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How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Histone deacetylase-2 and airway disease . The increased expression of inflammatory genes in inflammatory lung diseases is regulated by acetylation of core histones , whereas histone deacetylase-2 ( Q92769 ) suppresses inflammatory gene expression . Corticosteroids suppress inflammatory genes in asthma by inhibiting histone acetyltransferase and in particular by recruiting Q92769 to the nuclear factor-kappaB-activated inflammatory gene complex . This involves deacetylation of the acetylated glucocorticoid receptor . In P48444 , severe asthma and asthmatics who smoke , Q92769 is reduced , thus preventing corticosteroids from suppressing inflammation . The reduction in Q92769 appears to be secondary to increased oxidative and nitrative stress in the lungs . Antioxidants and inhibitors of nitric oxide synthesis may therefore restore corticosteroid sensitivity in P48444 , but this can also be achieved by low concentrations of theophylline and curcumin , which act as HDAC activators . DB00277 is a direct inhibitor of oxidant-activated phosphoinositide-3-kinase-delta , which is involved in inactivation of Q92769 . In the future selective O00329 inhibitors and more direct activators of Q92769 may be used to treat corticosteroid-resistant inflammatory diseases of the lung , including P48444 , severe asthma and asthma in smokers . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Environmental tobacco smoke exposure and airway hyperresponsiveness . Environmental tobacco smoke ( ETS ) exposure is a common health concern despite legislation to limit its presence , especially in public environments . ETS exposure is associated with changes in lung development and morphology , airway hyperresponsiveness and obstruction and development of asthma and its increased severity . However these effects of ETS exposure are not universally supported . Clinical data as well as studies in laboratory animals report ETS exposure may even attenuate airway hyperresponsiveness ( P35869 ) . Therefore , we lack complete understanding of ETS effects on pulmonary function as well as its mechanism of action . Disparate clinical and laboratory reports likely result from variables of ETS exposure , degrees of atopy and mechanisms of sensitization . The present review addresses the effects of ETS on P35869 reported in humans and animal models . ETS role as an adjuvant to P35869 as well as it contribution to development of antigenic tolerance is also reviewed . Possible neurogenic , cellular and intracellular mechanisms of ETS-induced Q5SW96 are proposed based on the existing literature . Enhanced understanding of the effects and mechanism of ETS will enhance therapy strategies in treatment of Q5SW96 and related disease such as P48444 as well as enhancing public presentation of convincing evidence to avoid ETS . A new cell culture-based assay quantifies vitamin K 2,3-epoxide reductase complex subunit 1 function and reveals warfarin resistance phenotypes not shown by the dithiothreitol-driven Q9BQB6 assay . BACKGROUND : DB00682 directly inhibits the vitamin K 2,3-epoxide reductase complex subunit 1 ( Q9BQB6 ) enzyme to effect anticoagulation . Q9BQB6 function has historically been assessed in vitro using a dithiothreitol ( DTT ) -driven vitamin K 2,3-epoxide reductase ( Q9BQB6 ) assay . DB00682 inhibits wild-type Q9BQB6 function by the DTT- Q9BQB6 assay . However , Q9BQB6 variants with warfarin resistance-associated missense mutations often show low Q9BQB6 activities and warfarin sensitivity instead of resistance . OBJECTIVES : A cell culture-based , indirect Q9BQB6 assay was developed and characterized that accurately reports warfarin sensitivity or resistance for wild-type and variant Q9BQB6 proteins . METHODS : Human coagulation factor (F)IX and Q9BQB6 variants were coexpressed in P29320 293T cells under standardized conditions at various warfarin concentrations . Secreted FIX activity served as surrogate marker to report wild-type and variant Q9BQB6 inhibition by warfarin . RESULTS AND CONCLUSIONS : DB00682 dose-response curves fit to the secreted FIX activity data for coexpressed hVKORC1 wild-type , Val29Leu , Val45Ala and Leu128Arg variants . The corresponding calculated IC50 values were 24.7 , 136.4 , 152.0 and 1226.4 nm , respectively . Basal activities in the absence of warfarin for all Q9BQB6 variants were similar to that of wild-type Q9BQB6 . Ranked IC50 values from the cell culture-based assay accurately reflect elevated warfarin dosages for patients with Q9BQB6 missense mutation-associated warfarin resistance . Leukemic challenge unmasks a requirement for O00329 in NK cell-mediated tumor surveillance . Specific inhibitors of PI3K isoforms are currently evaluated for their therapeutic potential in leukemia . We found that P11274 / P00519 (+) human leukemic cells express O00329 and therefore explored its impact on leukemia development . Using O00329 -deficient mice , we define a dual role of O00329 in leukemia . We observed a growth-promoting effect in tumor cells and an essential function in natural killer ( NK ) cell-mediated tumor surveillance : Abelson-transformed O00329 -deficient cells induced leukemia in P55895 -deficient mice with an increased latency , indicating that O00329 accelerated leukemia progression in vivo . However , the absence of O00329 also affected NK cell-mediated tumor surveillance . O00329 -deficient NK cells failed to lyse a large variety of target cells because of defective degranulation , as also documented by capacitance recordings . Accordingly , transplanted leukemic cells killed O00329 -deficient animals more rapidly . As a net effect , no difference in disease latency in vivo was detected if both leukemic cells and NK cells lack O00329 . Other tumor models confirmed that O00329 -deficient mice succumbed more rapidly when challenged with T- or B-lymphoid leukemic or B16 melanoma cells . Thus , the action of O00329 in the NK compartment is as relevant to survival of the mice as the delayed tumor progression . This dual function must be taken into account when using O00329 inhibitors as antileukemic agents in clinical trials . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Revelation of p53-independent function of Q13330 in DNA damage response via modulation of the P38936 P38936 -proliferating cell nuclear antigen pathway . Although metastasis-associated protein 1 ( Q13330 ) , a component of the nucleosome remodeling and deacetylase ( NuRD ) complex , is a DNA-damage response protein and regulates p53-dependent DNA repair , it remains unknown whether Q13330 also participates in p53-independent DNA damage response . Here , we provide evidence that Q13330 is a p53-independent transcriptional corepressor of P38936 ( P38936 ) , and the underlying mechanism involves recruitment of Q13330 -histone deacetylase 2 ( Q92769 ) complexes onto two selective regions of the P38936 ( P38936 ) promoter . Accordingly , Q13330 depletion , despite its effect on p53 down-regulation , superinduces P38936 ( P38936 ) , increases P38936 ( P38936 ) binding to proliferating cell nuclear antigen ( P12004 ) , and decreases the nuclear accumulation of P12004 in response to ionizing radiation . In support of a p53-independent role of Q13330 in DNA damage response , we further demonstrate that induced expression of Q13330 in p53-null cells inhibits P38936 ( P38936 ) promoter activity and P38936 ( P38936 ) binding to P12004 . Consequently , Q13330 expression in p53-null cells results in increased induction of gamma P16104 foci and DNA double strand break repair , and decreased DNA damage sensitivity following ionizing radiation treatment . These findings uncover a new target of Q13330 and the existence of an additional p53-independent role of Q13330 in DNA damage response , at least in part , by modulating the P38936 ( P38936 )- P12004 pathway , and thus , linking two previously unconnected NuRD complex and DNA-damage response pathways . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . The association between alveolar bone loss and pulmonary function : the VA Dental Longitudinal Study . The effect of oral conditions on medical outcomes is not well understood . The purpose of this epidemiological investigation was to examine whether the risk for chronic obstructive pulmonary disease ( P48444 ) is enhanced among individuals with a history of periodontal disease as assessed by radiographic alveolar bone loss ( P00519 ) . Subjects were selected from the VA Dental Longitudinal Study , a long-term study of aging and health in male veterans who were medically healthy at baseline . Subjects are not VA patients . Those subjects with a forced expiratory volume in 1 second ( FEV1 ) less than 65 % of predicted volume were categorized as having P48444 . P00519 was assessed by using full-mouth series periapical films measured by a Schei ruler . Bone loss at each interproximal site was measured in 20 % increments , and the mean whole-mouth bone loss score was calculated . Logistic regression analysis was used to determine the independent contribution of bone loss measurement at baseline to the subsequent risk of developing P48444 over a 25-year follow-up period . Covariates included measures of smoking , height , age , education , and alcohol consumption . Of the 1,118 medically healthy dentate men at baseline , 261 subsequently developed P48444 . We found that P00519 status at baseline was an independent risk factor for P48444 , with subjects in the worst population quintile of bone loss ( mean P00519 > 20 % per site ) found to be at significantly higher risk ( OR = 1.8 ; 95 % CI = 1.3 , 2.5 ) . The results of this analysis indicate that increased P00519 is associated with an increased risk for P48444 Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . P04150 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases . Airway epithelial cell PPARγ modulates cigarette smoke-induced chemokine expression and emphysema susceptibility in mice . Chronic obstructive pulmonary disease ( P48444 ) is a highly prevalent , chronic inflammatory lung disease with limited existing therapeutic options . While modulation of peroxisome proliferator-activating receptor ( Q07869 ) -γ activity can modify inflammatory responses in several models of lung injury , the relevance of the P37231 pathway in P48444 pathogenesis has not been previously explored . Mice lacking Pparg specifically in airway epithelial cells displayed increased susceptibility to chronic cigarette smoke ( CS ) -induced emphysema , with excessive macrophage accumulation associated with increased expression of chemokines , Ccl5 , Cxcl10 , and Cxcl15 . Conversely , treatment of mice with a pharmacological PPARγ activator attenuated Cxcl10 and Cxcl15 expression and macrophage accumulation in response to CS . In vitro , CS increased lung epithelial cell chemokine expression in a PPARγ activation-dependent fashion . The ability of PPARγ to regulate CS-induced chemokine expression in vitro was not specifically associated with peroxisome proliferator response element ( PPRE ) -mediated transactivation activity but was correlated with PPARγ-mediated transrepression of NF-κB activity . Pharmacological or genetic activation of PPARγ activity abrogated CS-dependent induction of NF-κB activity . Regulation of NF-κB activity involved direct PPARγ-NF-κB interaction and PPARγ-mediated effects on IKK activation , IκBα degradation , and nuclear translocation of p65 . Our data indicate that P37231 represents a disease-relevant pathophysiological and pharmacological target in P48444 . Its activation state likely contributes to NF-κB-dependent , CS-induced chemokine-mediated regulation of inflammatory cell accumulation . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . Genetic alterations and oncogenic pathways associated with breast cancer subtypes . Breast cancers can be divided into subtypes with important implications for prognosis and treatment . We set out to characterize the genetic alterations observed in different breast cancer subtypes and to identify specific candidate genes and pathways associated with subtype biology . mRNA expression levels of estrogen receptor , progesterone receptor , and P04626 were shown to predict marker status determined by immunohistochemistry and to be effective at assigning samples to subtypes . P04626 (+) cancers were shown to have the greatest frequency of high-level amplification ( independent of the P04626 amplicon itself ) , but triple-negative cancers had the highest overall frequencies of copy gain . Triple-negative cancers also were shown to have more frequent loss of phosphatase and tensin homologue and mutation of P06400 , which may contribute to genomic instability . We identified and validated seven regions of copy number alteration associated with different subtypes , and used integrative bioinformatics analysis to identify candidate oncogenes and tumor suppressors , including P04626 , Q14451 , O95251 , O15297 , P24385 , Q92769 , P55317 , and P20936 . We tested the candidate oncogene O95251 and showed that it enhances the anchorage-independent growth of breast cancer cells . The genome-wide and region-specific differences between subtypes suggest the differential activation of oncogenic pathways . Association of dopamine-related gene alleles , smoking behavior and decline in FEV1 in subjects with P48444 : findings from the lung health study . Cigarette smoking is the major risk factor for chronic obstructive pulmonary disease ( P48444 ) . Specific dopamine related gene alleles have previously been found to be associated with smoking initiation , maintenance and cessation . We investigated the association between specific dopamine related gene alleles and both change in smoking behavior and lung function change over time in individuals with mild-to-moderate P48444 . Subjects included a subset of participants in the Lung Health Study ( LHS ) , a smoking intervention study in smokers with mild to moderate P48444 . Smoking status was determined and lung function performed at baseline and annually for 5 years . In post-hoc analyses , we assessed the association of the dopamine receptor ( P14416 ) TaqI A1(+) allele ( A1A1 , A1A2 genotypes ) and A1(-) allele ( A2A2 genotype ) , and the dopamine transporter ( Q01959 ) 9R(+) allele ( 9R9R and 9R10R genotypes ) and 9R(-) allele ( 10R10R genotype ) with both changes in smoking status and lung function in a subset of LHS subjects . No significant associations were noted between variants in these genes and success in smoking cessation . However , in exploratory analyses that did not adjust for multiple comparisons , sustained male ( but not female ) quitters with the P14416 A1(-) allele and/or the Q01959 9R(+) allele showed an accelerated decline in Q99581 (1) similar to that of continuing smokers over 5 years after quitting smoking . These preliminary findings suggest that dopamine-related genes may play a role in the progression of P48444 , at least in the subset of male ex-smokers whose disease continues to progress despite sustained quitting , and warrants additional confirmatory and mechanistic studies . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Zinc-fingers and homeoboxes ( ZHX ) 2 , a novel member of the ZHX family , functions as a transcriptional repressor . Zinc-fingers and homeoboxes ( ZHX ) 1 is a transcription factor that interacts with the activation domain of the A subunit of nuclear factor-Y ( P23511 ) . Using a yeast two-hybrid system , a novel ubiquitous transcription factor Q9Y6X8 as a Q9UKY1 -interacting protein was cloned . Q9Y6X8 consists of 837 amino acid residues and contains two zinc-finger motifs and five homeodomains ( HDs ) as well as Q9UKY1 . The mRNA is expressed among various tissues . Q9Y6X8 not only forms a heterodimer with Q9UKY1 , but also forms a homodimer . Moreover , Q9Y6X8 interacts with the activation domain of P23511 . Further analysis revealed that Q9Y6X8 is a transcriptional repressor that is localized in the nuclei . Since Q9Y6X8 shares a number of properties in common with Q9UKY1 , we conclude that all these come under the ZHX family . The minimal functional domains of Q9Y6X8 were then characterized . The dimerization domain with both Q9UKY1 and Q9Y6X8 is the region containing HD1 , the domain that interacts with P23511 is the HD1 to Q92769 region , the repressor domain is the HD1 to a proline-rich region . Lastly , using an immunoprecipitation assay , we showed that Q9Y6X8 intrinsically interacts with P23511 in P29320 -293 cells and that Q9Y6X8 represses the promoter activity of the cdc25C gene stimulated by NF-Y in Drosophila Schneider line 2 cells . Thus the ZHX family of proteins may participate in the expression of a number of NF-Y-regulated genes via a more organized transcription network . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation .

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interacts_with DB06273?

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Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . Anti- P05231 -receptor-alpha ( tocilizumab ) does not inhibit human monocyte-derived dendritic cell maturation or alloreactive T-cell responses . Significant comorbidites and lethality complicate GVHD and its treatment . Targeting the cytokine milieu may improve GVHD control ; and P05231 is an attractive candidate , given its role in dendritic cell activation and T-cell differentiation . DB06273 is a humanized mAb to P05231 -receptor-α ( P08887 -α ) , which is Food and Drug Administration-approved for treatment of rheumatoid arthritis . Mouse transplant models have demonstrated that P05231 blockade also improves GVHD scores and survival . Definitive immunologic effects of P05231 inhibition have not emerged given inconsistent alterations in regulatory T cells ( Tregs ) and suppression of T-cell proliferation . Despite on-target suppression of P08887 -α signaling in human monocyte-derived dendritic cells ( moDCs ) and T cells , our data show no effect on moDC maturation/activation , alloreactive T-cell proliferation , Treg expansion , or allogeneic Th1/Th17 responses in vitro . These findings merit attention in any clinical trials of tocilizumab for GVHD prevention or treatment and provide a rationale for evaluating more specific inhibitors of downstream O60674 / P40763 signaling as well . Effect of tumor necrosis factor family member O43557 ( O43557 ) on the activation of basophils and eosinophils interacting with bronchial epithelial cells . Allergic asthma can cause airway structural remodeling , involving the accumulation of extracellular matrix and thickening of smooth muscle . P01375 ( P01375 ) family ligand O43557 ( O43557 ) is a cytokine that binds herpesvirus entry mediator ( Q92956 ) / Q92956 and lymphotoxin β receptor ( LTβR ) . O43557 induces asthmatic cytokine P35225 and fibrogenic cytokine transforming growth factor-β release from allergic asthma-related eosinophils expressing Q92956 and alveolar macrophages expressing LTβR , respectively , thereby playing crucial roles in asthmatic airway remodeling . In this study , we investigated the effects of O43557 on the coculture of human basophils/eosinophils and bronchial epithelial BEAS-2B cells . The expression of adhesion molecules , cytokines/chemokines , and matrix metalloproteinases ( MMP ) was measured by flow cytometry , multiplex , assay or ELISA . Results showed that O43557 could significantly promote intercellular adhesion , cell surface expression of intercellular adhesion molecule-1 , release of airway remodeling-related P05231 , P10145 , and P14780 from BEAS-2B cells upon interaction with basophils/eosinophils , probably via the intercellular interaction , cell surface receptors Q92956 and LTβR on BEAS-2B cells , and extracellular signal-regulated kinase , p38 mitogen activated protein kinase , and NF-κB signaling pathways . The above results , therefore , enhance our understanding of the immunopathological roles of O43557 in allergic asthma and shed light on the potential therapeutic targets for airway remodeling . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . DB06273 in pediatric rheumatology : the clinical experience . During the last two decades , clinical use of novel biological therapy has led to increased mechanistic understanding of complex rheumatological diseases . Conversely , basic and translational studies have led to development of new and varied therapeutic agents . These new medications which " target " specific steps in one or more immune pathways have the potential to control disease symptoms , improve quality of life and long-term prognosis , and perhaps in some , restore immunological tolerance . Use of these agents in clinical trials , combined with post-marketing surveillance , has revealed both the benefits and the undesirable side-effects of biological disease-modifying anti-rheumatic drugs ( DMARDs ) . In this review we focus on the use of tocilizumab , a monoclonal antibody directed against the P05231 receptor ( P08887 ) , which potently inhibits P05231 / P08887 signaling . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Differential effects of endotoxin and fibrinogen degradation products ( P14324 ) on liver synthesis of fibrinogen and albumin : evidence for the involvement of a novel monokine in the stimulation of fibrinogen synthesis induced by P14324 . 1. Administration of endotoxin or fibrinogen degradation products ( FDPs ) in rats increase fibrinogen synthesis comparable to that found during the acute phase response . 2 . An increased fibrinogen synthesis is also found in co-cultures of hepatocytes with peripheral blood mononuclear cells upon administration of endotoxin or FDPs , but not in primary cultures of hepatocytes alone . 3 . However , the increased synthesis of fibrinogen by FDPs is not accompanied by a decreased albumin synthesis , as in the case of stimulated fibrinogen synthesis induced by endotoxin in vivo and in co-cultures of hepatocytes with peripheral blood mononuclear cells , or induced by monocytic products in vivo and in primary cultures of hepatocytes alone . 4 . Since IL-1 and/or P05231 could not be accounted for the stimulation of fibrinogen synthesis without a decreased albumin synthesis , a novel monokine produced by mononuclear cells upon Q9NRC9 administration might be involved . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . P18509 , interleukin-6 and glucocorticoids regulate the release of vascular endothelial growth factor in pituitary folliculostellate cells . There is increasing evidence that hormones play an important role in the control of endothelial cell function and growth by regulating the production of vascular endothelial growth factor ( P15692 ) . P15692 regulates vascular permeability and represents the most powerful growth factor for endothelial cells . In the normal anterior pituitary , P15692 has been detected only in folliculostellate ( FS ) cells . In the present study , the regulation of the release of P15692 from FS-like mouse TtT/GF cells , and from FS cells of rat pituitary monolayer cell cultures was investigated using a specific P15692 ELISA . Basal release of P15692 was demonstrated in cultures of both TtT/GF cells and rat pituitary cells . Interestingly , the P15692 secretion was stimulated by both forms of pituitary adenylate cyclase-activating polypeptide ( PACAP-38 and PACAP-27 ) , indicating that this hypothalamic peptide regulates endothelial cell function and growth within the pituitary . P15692 secretion was also stimulated by interleukin-6 ( P05231 ) whereas basal , P05231 - and PACAP-stimulated secretion was inhibited by the synthetic glucocorticoid dexamethasone . The inhibitory action of dexamethasone was reversed by the glucocorticoid receptor antagonist DB00834 , suggesting that in FS cells functional glucocorticoid receptors mediate the inhibitory action of glucocorticoids on the P15692 secretion . The endocrine and auto-/paracrine control of P15692 production in pituitary FS cells by PACAP , P05231 and glucocorticoids may play an important role both in angiogenesis and vascular permeability regulation within the pituitary under physiological and pathophysiological conditions . In vitro and in vivo evidence for a role of the Q99572 receptor in the release of P01584 in the murine brain . The P2X(7) receptor ( P2X(7)R ) is a purinoceptor expressed predominantly by cells of immune origin , including microglial cells . P2X(7)R has a role in the release of biologically active proinflammatory cytokines such as P01584 , P05231 and TNFalpha . Here we demonstrate that when incubated with lipopolysaccharide ( LPS ) , glial cells cultured from brain of P2X(7)R(-/-) mice produce less P01584 compared to glial cells from brains of wild-type mice . This is not the case for TNFalpha and P05231 . Our results indicate a selective effect of the P2X7R gene deletion on release of P01584 release but not of P05231 and TNFalpha . In addition , we confirm that only microglial cells produce IL-1beta , and this release is dependent on P2X(7)R and ABC1 transporter . Because P01584 is a key regulator of the brain cytokine network and P2X(7)R is an absolute requirement for P01584 release , we further investigated whether response of brain cytokines to LPS in vivo was altered in P2X(7)R(-/-) mice compared to wild-type mice . P01584 and TNFalpha mRNAs were less elevated in the brain of P2X(7)R(-/-) than in the brain of wild-type mice in response to systemic LPS . These results show that P2X7R plays a key role in the brain cytokine response to immune stimuli , which certainly applies also to cytokine-dependent alterations in brain functions including sickness behavior . Biological activity of (lipo)polysaccharides of the exopolysaccharide-deficient mutant Rt120 derived from Rhizobium leguminosarum bv. trifolii strain Q96RJ0 . Lipopolysaccharides ( LPS ) from Rhizobium leguminosarum biovar trifolii Q96RJ0 ( RtTA1 ) and its mutant Rt120 in the pssBpssA intergenic region as well as degraded polysaccharides ( DPS ) derived from the LPS were elucidated in terms of their chemical composition and biological activities . The polysaccharide portions were examined by methylation analysis , MALDI-TOF mass spectrometry , and (1)H NMR spectroscopy . A high molecular mass carbohydrate fraction obtained from Rt120 DPS by Sephadex G-50 gel chromatography was composed mainly of L-rhamnose , 6-deoxy-L-talose , D-galactose , and D-galacturonic acid , whereas that from RtTA1 DPS contained L-fucose , 2-acetamido-2,6-dideoxy-D-glucose , D-galacturonic acid , 3-deoxy-3-methylaminofucose , D-glucose , D-glucuronic acid , and heptose . Relative intensities of the major (1)H NMR signals for O-acetyl and N-acetyl groups were 1 : 0.8 and 1 : 1.24 in DPS of Rt120 and RtTA1 , respectively . The intact mutant LPS exhibited a twice higher lethal toxicity than the wild type LPS . A higher in vivo production of TNFα and P05231 after induction of mice with Rt120 LPS correlated with the toxicity , although the mutant LPS induced the secretion of IL-1β and IFNγ more weakly than RtTA1 LPS . A polysaccharide obtained by gel chromatography on Bio-Gel P-4 of the high molecular mass material from Rt120 had a toxic effect on tumor HeLa cells but was inactive against the normal human skin fibroblast cell line . The polysaccharide from RtTA1 was inactive against either cell line . The potent inhibitory effect of the mutant DPS on tumor HeLa cells seems to be related with the differences in sugar composition . P05231 -receptor polymorphisms rs12083537 , rs2228145 , and rs4329505 as predictors of response to tocilizumab in rheumatoid arthritis . DB06273 ( TCZ ) , a monoclonal antibody targeting the human interleukin-6-receptor ( IL-6R ) , is indicated for the treatment of rheumatoid arthritis ( RA ) . We examined whether three P08887 single-nucleotide polymorphisms rs12083537 , rs2228145 ( formerly rs8192284 ) , and rs4329505 with previously reported functional effects were associated with clinical response to TCZ in a retrospective study cohort consisting of 79 RA patients . Three months after initiation of TCZ therapy , changes in swollen joint count ( SJC ) and , subordinately , tender joint count ( TJC ) , serum-CRP , DAS28-CRP , and EULAR-response were tested for association with the P08887 -haplotype or genotype . The major allele ( A ) of rs12083537 and the minor allele ( C ) of rs4329505 were associated with a poor SJC response ( P=0.02 and 0.02 , respectively ) . Moreover , the AAC-haplotype ( for rs12083537 , rs2228145 , and rs4329505 , respectively ) was associated with a poor SJC response ( P=0.00004 ) and , with borderline significance , EULAR-response ( P=0.05 ) . These data suggest that genetic variation in P08887 may aid in predicting TCZ therapy outcome in RA patients . Matrix metalloproteinase 9 ( gelatinase B ) is expressed in multinucleated giant cells of human giant cell tumor of bone and is associated with vascular invasion . Human giant cell tumor ( GCT ) consists of multinucleated giant cells and mononuclear stromal cells , and is characterized by frequent vascular invasion without distant metastases . To study the role of matrix metalloproteinases ( MMPs ) in the vascular invasion , we examined production of P03956 ( tissue collagenase ) , -2 ( gelatinase A ) , -3 ( stromelysin-1 ) , -9 ( gelatinase B ) , and tissue inhibitors of metalloproteinases ( P01033 and -2 ) in GCT . P14780 was highly and predominantly expressed in giant cells by both immunohistochemistry and in situ hybridization . Expression of other MMPs was also observed in some cases but was inconstant . Sandwich enzyme immunoassays demonstrated that P14780 is the predominant MMP secreted by GCT . There was a definite imbalance between the amounts of P14780 and those of TIMPs in the culture media of GCT , leading to detectable gelatinolytic activity in an assay using 14C-gelatin . Gelatin zymography demonstrated the main activity at about 90 kd , which was identified as the zymogen of P14780 by immunoblotting . Immunohistochemistry for type IV collagen and laminin , major basement membrane components , showed that disappearance of the proteins is closely associated with P14780 -positive giant cells . These results indicate the production of P14780 by multinucleated giant cells and suggest that the metalloproteinase may contribute to proteolysis associated with vascular invasion and local bone resorption in human GCT . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation .

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DB05812 acetate : a review of its use in patients with metastatic castration-resistant prostate cancer . DB05812 acetate ( Zytiga(®) ) is an orally administered , selective inhibitor of the 17α-hydroxylase and C17,20-lyase enzymatic activities of cytochrome P450 ( CYP ) 17 . P05093 is required for androgen biosynthesis , with androgen receptor signalling crucial in the progression from primary to metastatic prostate cancer . DB05812 acetate is approved in the European Union and the US , in combination with prednisone or prednisolone , for the treatment of men with metastatic castration-resistant prostate cancer ( CRPC ) . When administered in combination with prednisone in a placebo-controlled , multinational phase III study , abiraterone acetate significantly prolonged overall survival and radiographic progression-free survival ( rPFS ) in men with metastatic CRPC who had previously received docetaxel . In men with metastatic CRPC who had not previously received chemotherapy participating in a placebo-controlled , multinational phase III study , there was a strong trend towards an overall survival benefit , a significant prolongation in rPFS and significant delays in clinical decline , the need for chemotherapy and the onset of pain observed . Given the nature of the therapy , the overall tolerability profile of abiraterone acetate , in combination with prednisone , was acceptable in men with metastatic CRPC . DB05812 acetate is associated with hypokalaemia , hypertension , and fluid retention or oedema , secondary to its mechanism of action , and with cardiac adverse events and hepatotoxicity ; however , in the phase III studies the incidences of the most frequently reported grade 3 or 4 adverse events of special interest were relatively low . Although the final overall survival data in men with metastatic CRPC who have not previously received chemotherapy are awaited , current evidence indicates that abiraterone acetate is a useful option for the treatment of metastatic CRPC . Medical strategies for treatment of castration resistant prostate cancer ( CRPC ) docetaxel resistant . Current landscape of treatment of castration-resistant prostate cancer ( CRPC ) has recently changed . DB06772 , a new taxane with potential antineoplastic activity , has been approved by Food and Drug Administration ( FDA ) after docetaxel failure . In a phase III trial , cabazitaxel showed increased overall survival ( OS ) compared with mitoxantrone ( 15.1 vs. 12.7 mo , HR 0.70 , 95 % CI 0.59-0.83 , p < 0.0001 ) . Furthermore , chemotherapy is not the only strategy available : several studies have shown as CRPC remains dependent on androgen receptor function for growth . DB05812 acetate , an irreversible inhibitor of P05093 , has also been approved by FDA after docetaxel failure . In a phase III trial comparing abiraterone acetate to placebo , abiraterone showed improvement in OS ( 14.8 vs. 10.4 mo , HR 0.65 , 95 % CI 0.54-0.77 ; p < 0.0001 ) . This review will discuss current options and the ongoing trials for second-line treatment of CRPC including chemotherapy , hormonal therapies , antiangiogenetic and immune strategies . A-ring modified steroidal azoles retaining similar potent and slowly reversible P05093 inhibition as abiraterone . DB05812 acetate is a potent inhibitor of human cytochrome P450c17 ( P05093 , 17α-hydroxylase/17,20-lyase ) and is clinically used in combination with prednisone for the treatment of castration-resistant prostate cancer . Although many studies have documented the potency of abiraterone ( Abi ) in a variety of in vitro and in vivo systems for several species , the exact potency of Abi for human P05093 enzyme has not yet been determined , and the structural requirements for high-potency steroidal azole inhibitors are not established . We synthesized 4 Abi analogs differing in the A-B ring substitution patterns : 3α-hydroxy-Δ(4)-Abi ( 13 ) , 3-keto-Δ(4)-Abi ( 11 ) , 3-keto-5α-Abi ( 6 ) , and 3α-hydroxy-5α-Abi ( 5 ) . We measured the spectral binding constants ( Ks ) using purified and modified human P05093 along with the determination constants ( Ki ) applying a native human P05093 enzyme in yeast microsomes for these compounds as well as for ketoconazole . For Abi , 3-keto-Δ(4)-Abi , 3-keto-5α-Abi , and 3α-hydroxy-5α-Abi , the type 2 spectral changes gave the best fit for a quadratic equation , since in these experiments Ks values were 0.1-2.6nM , much lower than that for ketoconazole and 3α-hydroxy-Δ(4)-Abi ( Ks values were 140 and 1660nM , respectively ) . Inhibition experiments showed mixed inhibition patterns with Ki values of 7-80nM . Abi dissociation from the P05093 -Abi complex was incomplete and slow ; the t1/2 for dissociation was 1.8h , with 55 % of complex remaining after 5h . We conclude that Abi and the 3 related steroidal azoles ( 3-keto-Δ(4)-Abi , 3-keto-5α-Abi , and 3α-hydroxy-5α-Abi ) , which also mimic natural substrates , are extraordinarily potent inhibitors of human P05093 , whereas the 3α-hydroxy-Δ(4)-Abi is moderately potent and comparable to ketoconazole . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Use of prednisone with abiraterone acetate in metastatic castration-resistant prostate cancer . DB05812 acetate , a prodrug of the P05093 inhibitor abiraterone that blocks androgen biosynthesis , is approved for treatment of patients with metastatic castration-resistant prostate cancer ( mCRPC ) in combination with prednisone or prednisolone 5 mg twice daily . This review evaluates the basis for the effects of prednisone on mineralocorticoid-related adverse events that arise because of P05093 inhibition with abiraterone . Coadministration with the recommended dose of glucocorticoid compensates for abiraterone-induced reductions in serum cortisol and blocks the compensatory increase in adrenocorticotropic hormone seen with abiraterone . Consequently , 5 mg prednisone twice daily serves as a glucocorticoid replacement therapy when coadministered with abiraterone acetate , analogous to use of glucocorticoid replacement therapy for certain endocrine disorders . We searched PubMed to identify safety concerns regarding glucocorticoid use , placing a focus on longitudinal studies in autoimmune and inflammatory diseases and cancer . In general , glucocorticoid-related adverse events , including bone loss , immunosuppression , hyperglycemia , mood and cognitive alterations , and myopathy , appear dose related and tend to occur at doses and/or treatment durations greater than the low dose of glucocorticoid approved in combination with abiraterone acetate for the treatment of mCRPC . Although glucocorticoids are often used to manage tumor-related symptoms or to prevent treatment-related toxicity , available evidence suggests that prednisone and dexamethasone might also offer modest therapeutic benefit in mCRPC . Given recent improvements in survival achieved for mCRPC with novel agents in combination with prednisone , the risks of these recommended glucocorticoid doses must be balanced with the benefits shown for these regimens . Highly-selective 4-(1,2,3-triazole)-based P450c17a 17,20-lyase inhibitors . The orally-active P05093 inhibitor abiraterone acetate ( AA ) decreases adrenal and intratumoral androgen biosynthesis and is an effective agent for the treatment of prostate cancer . DB05812 potently inhibits both reactions catalyzed by P05093 , the 17α-hydroxylase ( hydroxylase ) reaction as well as the 17,20-lyase ( lyase ) transformation . P05093 hydroxylase inhibition prevents the synthesis of adrenal glucocorticoids and causes an accumulation of circulating mineralocorticoids . As a consequence of potent P05093 hydroxylase inhibition ( i.e. , lack of lyase selectivity ) , AA must be co-administered with the cortisol replacement prednisone and patients may experience the effects of mineralocorticoid excess syndrome ( MES ) . Herein , we describe rationally-designed , P05093 lyase-selective inhibitors that could prove safer and more effective than abiraterone . Using proprietary methodology , the high-affinity pyridine or imidazole metal-binding group found in current clinical P05093 inhibitors was replaced with novel , less avid , metal-binding groups in concert with potency-enhancing molecular scaffold modifications . This process produced a unique series of P05093 lyase-selective inhibitors that included the oral agent 6 ( VT-464 ) , now in Phase 2 prostate cancer clinical trials . The chemical methodology described is potentially applicable to the design of new and more effective metalloenzyme inhibitor treatments for a broad array of diseases . DB00563 in pediatric osteosarcoma : response and toxicity in relation to genetic polymorphisms and dihydrofolate reductase and reduced folate carrier 1 expression . OBJECTIVE : To determine the influence of the genotype and the level of expression of different enzymes involved in folate metabolism on the response to and toxicity of high-dose methotrexate treatment in pediatric osteosarcomas . STUDY DESIGN : P00374 and Reduced folate carrier 1 ( RFC1 ) semiquantitative expression was analyzed in 34 primary and metastatic osteosarcoma tissues by real-time polymerase chain reaction . The following polymorphisms were also analyzed in peripheral blood from 96 children with osteosarcoma and 110 control subjects : C677T , A1298C ( P42898 ) , G80A ( RFC1 ) , A2756G ( Q99707 ) , C1420T ( SHMT ) , the 28bp-repeat polymorphism , and 1494del6 of the P04818 gene . Treatment toxicity was scored after each cycle according to criteria from the World Health Organization . RESULTS : P00374 and RFC1 expression was lower in initial osteosarcoma biopsy specimens than in metastases ( P = .024 and P = .041 , respectively ) . RFC1 expression was moderately decreased in samples with poor histologic response to preoperative treatment ( P = .053 ) . Patients with osteosarcoma with P46379 /G4 hematologic toxicity were more frequently TT than CT/CC for C677T/ P42898 ( P = .023 ) and GG for A2756G/ Q99707 ( P = .048 and P = .057 for gastrointestinal and hematologic toxicity , respectively ) . CONCLUSIONS : The role of C677T/ P42898 and A2756G/ Q99707 on chemotherapy-induced toxicity should be further investigated in pediatric osteosarcomas receiving high-dose methotrexate . Altered expression of P00374 and RFC1 is a feasible mechanism by which osteosarcoma cells become resistant to methotrexate . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Recent progress in pharmaceutical therapies for castration-resistant prostate cancer . Since 2010 , six drugs have been approved for the treatment of castration-resistant prostate cancer , i.e. , P05093 inhibitor DB05812 , androgen receptor antagonist DB08899 , cytotoxic agent DB06772 , vaccine Sipuleucel-T , antibody DB06643 against receptor activator of nuclear factor kappa B ligand and radiopharmaceutical Alpharadin . All these drugs demonstrate improvement on overall survival , expect for DB06643 , which increases the bone mineral density of patients under androgen deprivation therapy and prolongs bone-metastasis-free survival . Besides further P05093 inhibitors ( Orteronel , Galeterone , VT-464 and CFG920 ) , androgen receptor antagonists ( ARN-509 , ODM-201 , AZD-3514 and EZN-4176 ) and vaccine Prostvac , more drug candidates with various mechanisms or new indications of launched drugs are currently under evaluation in different stages of clinical trials , including various kinase inhibitors and platinum complexes . Some novel strategies have also been proposed aimed at further potentiation of antitumor effects or reduction of side effects and complications related to treatments . Under these flourishing circumstances , more investigations should be performed on the optimal combination or the sequence of treatments needed to delay or reverse possible resistance and thus maximize the clinical benefits for the patients . Novel P05093 inhibitors : synthesis , biological evaluation , structure-activity relationships and modelling of methoxy- and hydroxy-substituted methyleneimidazolyl biphenyls . Recently , the steroidal P05093 inhibitor DB05812 entered phase II clinical trial for the treatment of androgen-dependent prostate cancer . As 17alpha-hydroxylase-17,20-lyase ( P05093 ) catalyzes the last step in androgen biosynthesis , inhibition of this target should affect not only testicular but also adrenal androgen formation . Therefore P05093 inhibitors should be advantageous over existing therapies , for example with DB00644 analogues . However , steroidal drugs are known for side effects which are due to affinities for steroid receptors . Therefore we decided to synthesize non-steroidal compounds mimicking the natural P05093 substrates pregnenolone and progesterone . The synthesis and biological evaluation of a series of 15 novel and highly active non-steroidal P05093 inhibitors are reported . The compounds were prepared via Suzuki-cross-coupling , Grignard reaction and CDI-assisted S(N)t-reaction with imidazole and their inhibitory activity was examined with recombinant human P05093 expressed in Escherichia coli . Promising compounds were further tested for their selectivity against the hepatic enzyme P08684 and the glucocorticoid-forming enzyme P15538 . All compounds turned out to be potent P05093 inhibitors . The most active compounds 7 and 8 were much more active than Ketoconazole showing activity comparable to DB05812 ( IC(50) values of 90 and 52nM vs. 72nM ) . Most compounds also showed higher selectivities than Ketoconazole , but turned out to be less selective than DB05812 . Docking studies using our P05093 protein model were performed with selected compounds to study the interactions between the inhibitors and the amino acid residues of the active site . Limited in vitro efficacy of P05093 inhibition on human castration resistant prostate cancer . Although accumulating evidence indicates high expression of P05093 (P45017A1) allows castration resistant prostate cancer ( CRPC ) to maintain high intratumoral androgen levels , the potential P45017A1 activity has not been characterized yet . The aim of this study was to examine the potential P05093 activity including 17α-hydroxylase and 17,20-lyase activities in human CRPC and the effect of a CYP17A inhibitor . We used three human CRPC cell lines : C4-2 and C4-2AT6 which was established from C4-2 under androgen ablation conditions for 6months , and PC3 . To ascertain the potential P05093 activity , we cultured with the steroid precursors : (13)C-[2,3,4]-progesterone ( 13C-Prog ) , and analyzed the sequential biosynthesis (13)C-[2,3,4]-17-hydroxyprogesterone ( 13C-17OHP ) and (13)C-[2,3,4]-androstenedione(13C-Adione) by liquid chromatography/mass spectrometry (LC/MS/MS).The C4-2AT6 cells showed significantly higher P05093 expression than C4-2 cells ( p < 0.001 ) . LC/MS/MS analysis enabled us to detect the 13C-17-OHP and 13C-A-dione in these cell lines . The concentration ratio of 13C-Adione/13C-17OHP ( Adione-17OHP ratio ) , which is thought to reflect the differences between 17-hydroxylase and 17,20-lyase activities , was then determined . The Adione-17OHP ratio in C4-2AT6 cells was significantly higher than that of C4-2 cells ( p < 0.001 ) . DB05812 were able to inhibit the CYP17A activities , although abiraterone did not have anti-proliferative effects on C4-2 and C4-2AT6 cells at clinically achievable concentrations of < 1000nM in vitro . The present study clearly demonstrates CRPC have the dual activities of P05093 mediated by 17-hydroxylase activity and 17,20-lyase activity . DB05812 does n't have an in vitro anti-proliferative efficacy in CRPC cells , suggesting limited efficacy in vitro . Multiple endocytic signals in the C-terminal tail of the cystic fibrosis transmembrane conductance regulator . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -dependent protein kinase ( PKA ) -activated chloride channel that is localized to the plasma membrane and endosomal compartment . Endosomal targeting of P13569 is attributed to the DB00135 (1424)-based internalization signal , identified in the C-terminal tail of the channel . Mutation of the DB00135 (1424) residue could partly inhibit the endocytosis of P13569 and its association with the adapter protein P05549 . To reveal additional endosomal targeting signals , site-directed mutagenesis of both a chimaera , composed of a truncated form of interleukin 2 receptor alpha chain ( TacT ) and the C-terminal tail of P13569 ( Ct ) , and the full-length P13569 was performed . Morphological and functional assays revealed the presence of multiple internalization motifs at the C-terminus , consisting of a phenylalanine-based motif ( DB00120 (1413) ) and a bipartite endocytic signal , comprising a tyrosine ( DB00135 (1424) ) and a di- DB00149 -based ( DB00149 (1430)- DB00149 ) motif . Whereas the replacement of any one of the three internalization motifs with alanine prevented the endocytosis of the TacT-Ct chimaera , mutagenesis of DB00120 (1413)- DB00149 impaired the biosynthetic processing of P13569 , indicating that DB00120 (1413) is indispensable for the native structure of P13569 . In contrast , replacement of DB00149 (1430)- DB00149 - and DB00135 (1424)-based signals with alanine increased the cell-surface density of both the chimaeras and P13569 in an additive manner . These results suggest that the internalization of P13569 is regulated by multiple endocytic sorting signals . DB05812 inhibits 3β-hydroxysteroid dehydrogenase : a rationale for increasing drug exposure in castration-resistant prostate cancer . PURPOSE : Treatment with abiraterone ( abi ) acetate prolongs survival in castration-resistant prostate cancer ( CRPC ) . Resistance to abi invariably occurs , probably due in part to upregulation of steroidogenic enzymes and/or other mechanisms that sustain dihydrotestosterone ( DB02901 ) synthesis , which raises the possibility of reversing resistance by concomitant inhibition of other required steroidogenic enzymes . On the basis of the 3β-hydroxyl , Δ(5)-structure , we hypothesized that abi also inhibits 3β-hydroxysteroid dehydrogenase/isomerase ( 3βHSD ) , which is absolutely required for DB02901 synthesis in CRPC , regardless of origins or routes of synthesis . EXPERIMENTAL DESIGN : We tested the effects of abi on 3βHSD activity , androgen receptor localization , expression of androgen receptor-responsive genes , and CRPC growth in vivo . RESULTS : Abi inhibits recombinant 3βHSD activity in vitro and endogenous 3βHSD activity in LNCaP and LAPC4 cells , including conversion of [ (3)H ] -dehydroepiandrosterone ( DB01708 ) to Δ(4)-androstenedione , androgen receptor nuclear translocation , expression of androgen receptor-responsive genes , and xenograft growth in orchiectomized mice supplemented with DB01708 . Abi also blocks conversion of Δ(5)- DB01524 to testosterone by 3βHSD . Abi inhibits 3βHSD1 and 3βHSD2 enzymatic activity in vitro ; blocks conversion from DB01708 to androstenedione and DB02901 with an IC(50) value of less than 1 μmol/L in CRPC cell lines ; inhibits androgen receptor nuclear translocation ; expression of O15393 , prostate-specific antigen , and Q13451 ; and decreases CRPC xenograft growth in DB01708 -supplemented mice . CONCLUSIONS : We conclude that abi inhibits 3βHSD-mediated conversion of DB01708 to active androgens in CRPC . This second mode of action might be exploited to reverse resistance to P05093 inhibition at the standard abi dose by dose-escalation or simply by administration with food to increase drug exposure . P05093 inhibitors in castration-resistant prostate cancer . The majority of prostate cancer ( PCa ) cases are diagnosed as a localized disease . Definitive treatment , active surveillance or watchful waiting are employed as therapeutic paradigms . The current standard of care for the treatment of metastatic PCa is either medical or surgical castration . Once PCa progresses in spite of castrate androgen levels it is termed ' castration-resistant prostate cancer ' ( CRPC ) . Patients may even exhibit rising PSA levels with possible bone , lymph node or solid organ metastases . In 2010 , the only agent approved for the treatment of CRPC was docetaxel , a chemotherapeutic agent . It is now known that cells from patients with CRPC express androgen receptors ( AR ) and remain continuously influenced by androgens . As such , treatments with novel hormonal agents that specifically target the biochemical conversion of cholesterol to testosterone have come to the forefront . The use of cytochrome P450c17 ( P05093 ) inhibitor underlies one of the most recent advances in the treatment of CRPC . DB05812 acetate ( AA ) was the first P05093 inhibitor approved in the United States . This review will discuss CRPC in general with a specific focus on AA and novel P05093 inhibitors . AA clinical trials will be reviewed along with other novel adjunct treatments that may enhance the effectiveness of abiraterone therapy . Furthermore , the most recently identified P05093 inhibitors Orteronel , Galeterone , VT-464 , and CFG920 will also be explored . DB05812 acetate : oral androgen biosynthesis inhibitor for treatment of castration-resistant prostate cancer . Prostate cancer is the second leading cause of cancer death in men in the US and Europe . The treatment of advanced-stage prostate cancer has been androgen deprivation . Medical castration leads to decreased production of testosterone and dihydrotestosterone by the testes , but adrenal glands and even prostate cancer tissue continue to produce androgens , which eventually leads to continued prostate cancer growth despite castrate level of androgens . This stage is known as castrate-resistant prostate cancer ( CRPC ) , which continues to be a challenge to treat . Addition of androgen antagonists to hormonal deprivation has been successful in lowering the prostate-specific antigen levels further , but has not actually translated into life-prolonging options . The results of several contemporary studies have continued to demonstrate activation of the androgen receptor as being the key factor in the continued growth of prostate cancer . Blockade of androgen production by nongonadal sources has led to clinical benefit in this setting . One such agent is abiraterone acetate , which significantly reduces androgen production by blocking the enzyme , cytochrome P450 17 alpha-hydroxylase ( P05093 ) . This has provided physicians with another treatment option for patients with CRPC . The landscape for prostate cancer treatment has changed with the approval of cabazitaxel , sipuleucel-T and abiraterone . Here we provide an overview of abiraterone acetate , its mechanism of action , and its potential place for therapy in CRPC . Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha-reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha-reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer(PCa) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or= +/-1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR; P24385 ) , and signal transduction ( P04626 ; V- P62158 ; Q07889 ) whereas , underexpressed genes ( KLK3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology . Effect of abiraterone acetate plus prednisone on the QT interval in patients with metastatic castration-resistant prostate cancer . PURPOSE : DB05812 is the active metabolite of the pro-drug abiraterone acetate ( AA ) and a selective inhibitor of P05093 , a key enzyme in testosterone synthesis , and improves overall survival in postdocetaxel metastatic castration-resistant prostate cancer ( mCRPC ) . This open-label , single-arm phase 1b study was conducted to assess the effect of AA and abiraterone on the QT interval . METHODS : The study was conducted in 33 patients with mCRPC . Patients received AA 1,000 mg orally once daily + prednisone 5 mg orally twice daily . Electrocardiograms ( ECGs ) were collected in triplicate using 12-lead Holter monitoring . Baseline ECGs were obtained on Cycle 1 Day-1 . Serial ECG recordings and time-matched pharmacokinetic ( PK ) blood samples were collected over 24 h on Cycle 1 Day 1 and Cycle 2 Day 1 . Serial PK blood samples were also collected over 24 h on Cycle 1 Day 8 . RESULTS : After AA administration , the upper bound of the 2-sided 90 % confidence interval ( CI ) for the mean baseline-adjusted QTcF change was < 10 ms ; no patients discontinued due to QTc prolongation or adverse events . No apparent relationship between change in QTcF and abiraterone plasma concentrations was observed [ estimated slope ( 90 % CI ) : 0.0031 ( -0.0040 , 0.0102 ) ] . CONCLUSIONS : There is no significant effect of AA plus prednisone on the QT/QTc interval in patients with mCRPC . Androgen synthesis inhibitors in the treatment of castration-resistant prostate cancer . Suppression of gonadal testosterone synthesis represents the standard first line therapy for treatment of metastatic prostate cancer . However , in the majority of patients who develop castration-resistant prostate cancer ( CRPC ) , it is possible to detect persistent activation of the androgen receptor ( AR ) through androgens produced in the adrenal gland or within the tumor itself . DB05812 acetate was developed as an irreversible inhibitor of the dual functional cytochrome P450 enzyme P05093 with activity as a 17α-hydroxylase and 17,20-lyase . P05093 is necessary for production of nongonadal androgens from cholesterol . Regulatory approval of abiraterone in 2011 , based on a phase III trial showing a significant improvement in overall survival ( OS ) with abiraterone and prednisone versus prednisone , represented proof of principle that targeting AR is essential for improving outcomes in men with CRPC . Inhibition of 17α-hydroxylase by abiraterone results in accumulation of upstream mineralocorticoids due to loss of cortisol-mediated suppression of pituitary adrenocorticotropic hormone ( DB01285 ) , providing a rationale for development of P05093 inhibitors with increased specificity for 17,20-lyase ( orteronel , galeterone and VT-464 ) that can potentially be administered without exogenous corticosteroids . In this article , we review the development of abiraterone and other P05093 inhibitors ; recent studies with abiraterone that inform our understanding of clinical parameters such as drug effects on quality-of-life , potential early predictors of response , and optimal sequencing of abiraterone with respect to other agents ; and results of translational studies providing insights into resistance mechanisms to P05093 inhibitors leading to clinical trials with drug combinations designed to prolong abiraterone benefit or restore abiraterone activity . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation . P19957 kinase p110β : a therapeutic target in advanced prostate cancers . Prostate cancers in the castration-resistant stage are life-threatening because they are not curable in clinic . The novel androgen receptor inhibitor Xandi ( DB08899 ) and the new P05093 inhibitor Zytiga ( DB05812 ) prolonged patient survival only a few months in advanced prostate cancers . Therefore , novel therapeutic agents for advanced prostate cancers are urgently needed . P19957 kinases are major intracellular signaling molecules that regulate multiple signal pathways related to cellular metabolism , cytokinesis , growth and survival . Accumulating evidence in the literature indicates that some isoforms of this kinase family are oncogenic and abnormally expressed in various human cancers , including prostate cancers . Recent extensive studies from our group and others showed that P19957 kinase p110β is aberrantly overexpressed in advanced prostate cancers and is critical for prostate cancer development and progression as demonstrated in cell-based and animal models . Importantly , novel p110β-specific inhibitors have been developed and are currently been testing in clinical trials . In this article , we will briefly summarize recent developments in this regard . DB05812 acetate : redefining hormone treatment for advanced prostate cancer . Prostate cancer has long since been recognised as being hormonally driven via androgen receptor signalling . DB05812 acetate ( AA ) is a rationally designed P05093 inhibitor that blocks the conversion of androgens from non-gonadal precursors effectively , thus reducing testosterone to undetectable levels . AA has recently been proved to extend survival for men with metastatic castration-resistant prostate cancer who have progressive disease after first-line chemotherapy treatment . In addition , it is currently being tested in a Phase III trial in the pre-chemotherapy setting . This paper will review the preclinical discovery and clinical development of AA and will outline the strategy of parallel translational research . New agents and strategies for the hormonal treatment of castration-resistant prostate cancer . IMPORTANCE OF THE FIELD : Hormonal therapy with medical or surgical castration is the mainstay of systemic therapy for advanced prostate cancer . Depletion of gonadal testosterone in circulation is typically initially effective , although responses are transient and metastatic disease progresses as castration-resistant prostate cancer ( CRPC ) . AREAS COVERED IN THIS REVIEW : CRPC is accompanied by a gain of function in the androgen receptor ( AR ) , which may occur at the level of AR itself or through intratumoral repletion of androgens that in turn stimulate AR . Investigational drugs in clinical trials have promising activity in CRPC . DB05812 acetate is a P05093 inhibitor that blocks the synthesis of adrenal androgens . MDV3100 is a nonsteroidal AR antagonist with a greater binding affinity than other AR antagonists currently in clinical use . Insights into the mechanisms of intratumoral steroidogenesis in CRPC have defined other potential targets . Metabolism from DB01708 to testosterone and dihydrotestosterone requires 3-hydroxyl oxidation and Delta(5) isomerization to Delta(4) by 3beta-hydroxysteroid dehydrogenase ( 3betaHSD ) and 17-keto reduction by 17beta-hydroxysteroid dehydrogenase ( 17betaHSD ) -3 or -5 . AR activation in CRPC by intratumoral steroids requires these enzymatic steps . Investigation into specific inhibitors of 3betaHSD and 17betaHSD are required to determine their efficacy and potential roles in the treatment of CPRC . WHAT THE READER WILL GAIN : Readers will gain an understanding of the biology of CRPC , new investigational hormonal agents and novel approaches to the treatment of CRPC . TAKE HOME MESSAGE : Intratumoral androgens drive CRPC progression . New investigational hormonal agents that inhibit intratumoral androgens are highly active in the treatment of CRPC . Alternative strategies hold the promise for the development of other agents with novel mechanisms of action . DB05812 acetate : targeting persistent androgen dependence in castration-resistant prostate cancer . DB05812 acetate is the first second-line hormonal agent proven to improve survival in metastatic castration-resistant prostate cancer . It selectively inhibits cytochrome P450 17 ( P05093 ) α-hydroxylase and cytochrome17,20 ( C17,20 ) -lyase , which are enzymes critical for androgen synthesis . DB05812 acetate was initially approved in the United States in 2011 after demonstrating a 4-month survival benefit in docetaxel-refractory metastatic prostate cancer . The FDA recently expanded its indication for use in the pre-chemotherapy setting after it elicited significant delays in disease progression and a strong trend for increased overall survival in phase III studies . Ongoing investigations of abiraterone are evaluating its efficacy in earlier disease states , exploring its synergy in combination with other therapeutic agents , and assessing the necessity for administration of concurrent steroids and gonadal suppression . The identification and development of predictive biomarkers will optimize the incorporation of abiraterone into the management of advanced prostate cancer . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Antitumor activity with P05093 blockade indicates that castration-resistant prostate cancer frequently remains hormone driven . DB05812 acetate is a potent , selective , and orally bioavailable small molecule inhibitor of P05093 , an enzyme that catalyzes two key serial reactions ( 17 alpha hydroxylase and 17,20 lyase ) in androgen and estrogen biosynthesis . Clinical trials have confirmed that specific inhibition of P05093 is safe and results in clinically important antitumor activity in up to 70 % of castrate patients with advanced prostate cancer resistant to currently available endocrine therapies . These clinical data indicate that castration-resistant prostate cancer frequently remains hormone dependent and has confirmed that this disease should no longer be described as " hormone resistant or refractory " . Biomarker studies , including the analysis of ETS gene fusion status , on patients treated with abiraterone acetate may allow enrichment of patients with a sensitive phenotype in future studies of therapeutics targeting P05093 . The role of the central flexible region on the aggregation and conformational properties of human ataxin-3 . Aggregation of human ataxin-3 ( P01008 ) into amyloid fibrils is responsible for spinocerebellar ataxia type 3 . This protein consists of a folded N-terminal domain ( Josephin domain , residues 1-182 ) , a central flexible region ( residues 183-291 ) , a poly-glutamine sequence of variable length and a short C-terminal flexible region . Very little is known about the influence of the central flexible region on the conformational and aggregation properties of this protein . The present study aimed to investigate the specific role of this portion of the protein ( residues 183-291 ) . Accordingly , protein fragments 1-182 ( P01008 /182 ) and 1-291 ( P01008 /291 ) were produced and compared by thioflavin-T fluorescence , Fourier transform infrared spectroscopy , CD , intrinsic fluorescence and P19957 -MS . It is shown that the central flexible region enhances protein aggregation and can populate conformational states with different degrees of compactness . Both monomeric and dimeric partially-folded forms are identified for both protein fragments under denaturing conditions . Partially-folded monomers and dimers accumulate to a larger extent in P01008 /291 . These species represent good candidates for early intermediates of the aggregation process under the experimental conditions employed in the present study . Interactions of abiraterone , eplerenone , and prednisolone with wild-type and mutant androgen receptor : a rationale for increasing abiraterone exposure or combining with MDV3100 . Prostate cancer progression can be associated with androgen receptor ( AR ) mutations acquired following treatment with castration and/or an antiandrogen . DB05812 , a rationally designed inhibitor of P05093 recently approved for the treatment of docetaxel-treated castration-resistant prostate cancer ( CRPC ) , is often effective , but requires coadministration with glucocorticoids to curtail side effects . Here , we hypothesized that progressive disease on abiraterone may occur secondary to glucocorticoid-induced activation of mutated AR . We found that prednisolone plasma levels in patients with CRPC were sufficiently high to activate mutant AR . P08235 antagonists , such as spironolactone and eplerenone that are used to treat side effects related to mineralocorticoid excess , can also bind to and activate signaling through wild-type or mutant AR . DB05812 inhibited in vitro proliferation and AR-regulated gene expression of AR-positive prostate cancer cells , which could be explained by AR antagonism in addition to inhibition of steroidogenesis . In fact , activation of mutant AR by eplerenone was inhibited by MDV3100 , bicalutamide , or greater concentrations of abiraterone . Therefore , an increase in abiraterone exposure could reverse resistance secondary to activation of AR by residual ligands or coadministered drugs . Together , our findings provide a strong rationale for clinical evaluation of combined P05093 inhibition and AR antagonism . Inhibition of the androgen receptor by mineralocorticoids at levels physiologically achieved in serum in patients treated with abiraterone acetate . BACKGROUND : DB05812 acetate ( AA ) , a highly potent P05093 inhibitor , has demonstrated marked clinical benefit in patients with metastatic castration-resistant prostate cancer ( CRPC ) . Phase I trials of AA without prednisone showed significant elevation of serum mineralocorticoid concentrations . The aim of this study was to elucidate the biological significance of elevated mineralocorticoid levels on androgen receptor ( AR ) activity in prostate cancer ( PC ) cells . METHODS : Fluorescence resonance energy transfer ( FRET ) assay was used to assess the effect of mineralocorticoids on androgen-induced conformational change of the AR . LAPC4 , LNCaP and LN-AR cells that were cultured and treated with androgens were exposed to mineralocorticoids at varying concentrations , including levels measured in the serum of AA-treated patients in a phase I trial . AR-dependent transcriptional activity and cell growth were measured in these cell lines to determine the biological impact of mineralocorticoids on PC cells . RESULTS : Corticosterone ( CS ) and deoxycorticosterone ( DOC ) inhibited androgen-induced conformational change of the AR in the FRET assay . CS inhibited AR-dependent transcriptional activity and cell growth at concentrations comparable to those measured in the serum of AA-treated patients . DOC inhibited AR transcriptional activity and cell growth at 10-fold greater concentrations than measured in the serum of AA-treated patients . CONCLUSIONS : Mineralocorticoids directly inhibit androgen-induced conformational change of the AR . CS inhibits AR transcriptional activity and PC cell growth at concentrations found in the serum of patients treated with AA . Further investigation of the potential therapeutic implications of mineralocorticoids in AA-treated CRPC patients is warranted . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . Battle of the kinases : integration of adrenal responses to DB02527 , DG and Ca2+ at the level of steroidogenic cytochromes P450 and 3betaHSD expression in H295R cells . While DB01285 receptors ( activating the protein kinase A pathway ) are expressed throughout the human/bovine/ovine zona glomerulosa ( zg ) and zona fasciculata ( zf ) , there are clear zonal differences in AII Type-1 receptor levels ( activating protein kinase C/Ca2+ ) , as well as resting membrane potential . Thus zg is most responsive to AII and K+ ( Ca2+ signalling ) , while zf is less responsive to AII with no K+ response . Zonal function in turn requires differential expression of P05093 /3betaHSD and P19099 / P15538 . We have used the H295R cell to study how differential activation of kinase A , kinase C and Ca2+/calmodulin ( P62158 ) kinases may alter the relative expression of the steroidogenic P450s and 3betaHSDII . While P05108 , P05093 , 3betaHSDII , P08686 , and P15538 are all induced by increases in DB02527 , studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively ( P08686 , 3betaHSDII ) or negatively ( P05093 , P05108 ) by protein kinase C . Thus adrenal 3betaHSDII and P08686 expression is high in zg and zf , but P05093 is not expressed in the zg where AII activation of kinase C is highest . In turn both K+ and AII-induced elevation of Ca2+ strongly induces P19099 but not P15538 , consistent with preferential expression of P19099 in the zg . We conclude that differential signaling through kinase C and P62158 kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones . Clinical and biochemical consequences of P05093 inhibition with abiraterone given with and without exogenous glucocorticoids in castrate men with advanced prostate cancer . CONTEXT : DB05812 acetate is a small-molecule cytochrome P450 17A1 ( P05093 ) inhibitor that is active in castration-resistant prostate cancer . OBJECTIVE : Our objective was to determine the impact of abiraterone with and without dexamethasone treatment on in vivo steroidogenesis . DESIGN AND METHODS : We treated 42 castrate , castration-resistant prostate cancer patients with continuous , daily abiraterone acetate and prospectively collected blood and urine before and during abiraterone treatment and after addition of dexamethasone 0.5 mg daily . RESULTS : Treatment with single-agent abiraterone acetate was associated with accumulation of steroids with mineralocorticoid properties upstream of P05093 . This resulted in side effects , including hypertension , hypokalemia , and fluid overload , in 38 of 42 patients that were generally treated effectively with eplerenone . Importantly , serum and urinary androgens were suppressed by more than 90 % from baseline . Urinary metabolites of 17-hydroxypregnenolone and 17-hydroxyprogesterone downstream of 17α-hydroxylase remained unchanged . However , 3α5α-17-hydroxypregnanolone , which can be converted via the backdoor pathway toward 5α-dihydrotestosterone , increased significantly and correlated with levels of the major 5α-dihydrotestosterone metabolite androsterone . In contrast , urinary metabolites of 11-deoxycortisol and active glucocorticoids declined significantly . Addition of dexamethasone to abiraterone acetate significantly suppressed DB01285 and endogenous steroids , including 3α5α-17-hydroxypregnanolone . CONCLUSION : P05093 inhibition with abiraterone acetate is characterized by significant suppression of androgen and cortisol synthesis . The latter is associated with a rise in DB01285 that causes raised mineralocorticoids , leading to side effects and incomplete 17α-hydroxylase inhibition . Concomitant inhibition of 17,20-lyase results in diversion of 17-hydroxyprogesterone metabolites toward androgen synthesis via the backdoor pathway . Addition of dexamethasone reverses toxicity and could further suppress androgens by preventing a rise in substrates of backdoor androgen synthesis . [ DB05812 acetate : a novel therapeutic option in hormone-refractory prostate cancer ] . Until recently , only therapy with docetaxel and prednisone has been shown to prolong survival in men with hormonorefractory metastatic prostate cancer . With approvals of sipuleucel-T , cabazitaxel , and abiraterone acetate , all based on improvement in overall survival , the scenary for management of men with metastatic prostate cancer has dramatically changed . DB05812 acetate was developed to specifically inhibit cytochrome P450 (CYP)17A1 , which is an essential enzyme in the biosynthesis of testosterone . In the phase III , the trial treatment with abiraterone acetate plus prednisone prolongs overall survival relative to prednisone alone in patients with metastatic castration-resistant prostate cancer who have disease progression after treatment with docetaxel and associated with an acceptable tolerability profile , which was generally similar to that of the placebo plus prednisone group . However , adverse events resulting from elevated mineralocorticoid levels because of P05093 inhibition , fluid retention and oedema , hypokalaemia , hypertension occurred in significantly more in abiraterone acetate plus prednisone than in placebo plus prednisone . P05093 inhibitors -- abiraterone , C17,20-lyase inhibitors and multi-targeting agents . As the first in class steroid 17α-hydroxylase/C17,20-lyase ( P05093 ) inhibitor , abiraterone acetate ( of which the active metabolite is abiraterone ) has been shown to improve overall survival in patients with castration-resistant prostate cancer ( CRPC ) -- in those who are chemotherapy-naive and those previously treated with docetaxel . Furthermore , the clinical success of abiraterone demonstrated that CRPC , which has previously been regarded as an androgen-independent disease , is still driven , at least in part , by androgens . More importantly , abiraterone is a ' promiscuous ' drug that interacts with a number of targets , which dictate its clinical benefits and adverse effects profile . Besides P05093 inhibition , abiraterone acts as an antagonist to the androgen receptor and inhibits 3β-hydroxysteroid dehydrogenase -- two effects that potentially contribute to its antitumour effects . However , the inhibition of the 17α-hydroxylase activity of P05093 , P15538 and a panel of hepatic CYP enzymes leads to adverse effects and toxicities that include secondary mineralocorticoid excess . DB05812 is also associated with increased incidence of cardiac disorders . Under such circumstances , development of new P05093 inhibitors as an additional line of defence is urgently needed . To achieve enhanced clinical benefits , new strategies are being explored that include selective inhibition of the C17,20-lyase activity of P05093 and multi-targeting strategies that affect androgen synthesis and signalling at different points . Some of these strategies-including the drugs orteronel , VT-464 and galeterone -- are supported by preclinical data and are being explored in the clinic . Prostate cancer-from steroid transformations to clinical translation . The survival benefit conferred by two hormonal agents in phase III trials has clinically validated the long suspected and now widely recognized phenomenon of castration-resistant prostate cancer ( CRPC ) hormone dependence . DB05812 inhibits steroid 17α-hydroxylase/17,20-lyase ( P05093 ) and blocks androgen synthesis , whereas enzalutamide directly binds and antagonizes the androgen receptor . Both agents are highly effective against CRPC and significantly prolong survival following docetaxel treatment . However , this clinical validation of the androgen pathway has led to questions regarding the fundamental mechanisms of CRPC , as well as resistance to abiraterone and enzalutamide . Our understanding of the predominant steroid transformation pathways that lead to dihydrotestosterone synthesis in CRPC is evolving . The role of steroidogenesis in the development of resistance to abiraterone and enzalutamide remains uncertain . The specific roles of candidate enzyme targets in the development of resistance to these agents must be defined if we are to identify novel targets for improved pharmacologic therapies . Is abiraterone acetate well tolerated and effective in the treatment of castration-resistant prostate cancer ? This Practice Point commentary discusses the findings of the first phase I trial to evaluate abiraterone acetate ( an inhibitor of the androgen-regulating enzyme P05093 ) in the treatment of castration-resistant prostate cancer . This open-label , dose-escalation study by Attard et al. showed that abiraterone was well tolerated but often induced a syndrome of secondary mineralocorticoid excess that improved with eplerenone ( a mineralocorticoid receptor antagonist ) . DB05812 is a potent suppressor of adrenal androgen synthesis , and produced lasting prostate-specific antigen responses in approximately half of the patients . A few patients had partial regression of distant metastases . Although promising , these results should be interpreted with caution owing to the small sample size and because the study was not primarily designed to examine drug efficacy . Multi-institutional , prospective trials should provide additional information on the tolerability and activity of this compound and further define the population most likely to benefit from this endocrine approach . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Ketoconazole inhibits the cellular uptake of anandamide via inhibition of FAAH at pharmacologically relevant concentrations . BACKGROUND : The antifungal compound ketoconazole has , in addition to its ability to interfere with fungal ergosterol synthesis , effects upon other enzymes including human P08684 , P05093 , lipoxygenase and thromboxane synthetase . In the present study , we have investigated whether ketoconazole affects the cellular uptake and hydrolysis of the endogenous cannabinoid receptor ligand anandamide ( AEA ) . METHODOLOGY/PRINCIPAL FINDINGS : The effects of ketoconazole upon endocannabinoid uptake were investigated using HepG2 , CaCo2 , PC-3 and P13671 cell lines . Fatty acid amide hydrolase ( FAAH ) activity was measured in HepG2 cell lysates and in intact P13671 cells . Ketoconazole inhibited the uptake of AEA by HepG2 cells and CaCo2 cells with IC50 values of 17 and 18 µM , respectively . In contrast , it had modest effects upon AEA uptake in PC-3 cells , which have a low expression of FAAH . In cell-free HepG2 lysates , ketoconazole inhibited FAAH activity with an IC50 value ( for the inhibitable component ) of 34 µM . CONCLUSIONS/SIGNIFICANCE : The present study indicates that ketoconazole can inhibit the cellular uptake of AEA at pharmacologically relevant concentrations , primarily due to its effects upon FAAH . Ketoconazole may be useful as a template for the design of dual-action FAAH/ P05093 inhibitors as a novel strategy for the treatment of prostate cancer . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . The 5α-androstanedione pathway to dihydrotestosterone in castration-resistant prostate cancer . The survival and progression of prostate cancer are generally dependent on expression of the androgen receptor ( AR ) , as well as the availability of endogenous AR agonists . Originating from the gonads , testosterone is released into circulation and is converted by steroid-5α-reductase in prostate cancer to 5α-dihydrotestosterone ( DB02901 ) , potently activating AR and driving tumor progression . Advanced prostate cancer is initially treated with gonadal testosterone depletion , which suppresses this cascade of events and typically leads to a treatment response . Eventually , resistance to testosterone deprivation occurs with " castration-resistant " prostate cancer ( CRPC ) and is driven by the intratumoral synthesis of DB02901 . The generation of DB02901 occurs in large part from adrenal 19-carbon precursor steroids , which are dependent on expression of P05093 . Although the path from adrenal precursor steroids to DB02901 was generally thought to require 5α-reduction of testosterone , recent data suggest that it instead involves conversion from Δ-androstenedione by steroid-5α-reductase isoenzyme-1 to 5α-androstanedione , followed by subsequent conversion to DB02901 . The 5α-androstanedione pathway to DB02901 therefore bypasses testosterone entirely . DB05812 acetate effectively inhibits P05093 , blocks the synthesis of androgens , and extends the survival of men with CRPC . Further progress in the hormonal treatment of CRPC is dependent on an understanding of the mechanisms that underlie CRPC and resistance to abiraterone acetate . [ DB05812 acetate(ZYTIGA®)-development and literature review ] . DB05812 acetate(AA)has been approved in more than 80 countries for the treatment of patients with metastatic castration-resistant prostate cancer(mCRPC) . In July 2013 , a marketing approval application for AA was submitted to the Japanese Ministry of Health , Labour , and Welfare . AA is a selective inhibitor of P05093 , a crucial enzyme for androgen biosynthesis . AA exerts its anti-tumor activity by directly inhibiting androgen production at all three sources , i. e. , the testes , adrenal glands , and tumor itself . Data from international phase III studies and phase I and II studies in Japan have indicated that AA improves the overall survival and quality of life(QoL)of patients with mCRPC . Herein , we have summarized the development of AA and the results of important international and local clinical trials in Japan . In addition , the effect of food on AA bioavailability , concomitant steroid use , and liver function test abnormalities have been discussed regarding the appropriate use of AA . DB05812 acetate : a promising drug for the treatment of castration-resistant prostate cancer . DB05812 acetate ( CB7630 ) , a pregnenolone analog , is an orally administered small molecule that irreversibly inhibits a rate-limiting enzyme in androgen biosynthesis , P05093 , and blocks the synthesis of androgens in the testes , adrenal glands and prostate without causing adrenal insufficiency . In clinical studies , abiraterone acetate is well tolerated and shows promising clinical activity in castration-resistant prostate cancer . The recommended Phase II dose of abiraterone acetate is 1000 mg orally daily in combination with prednisone 5 mg twice daily . Side effects are minimal and mostly associated with secondary mineralocorticoid excess , owing to a compensatory increase in upstream steroids , such as deoxycorticosterone and corticosterone . These include hypertension , hypokalemia and edema and are easily manageable with a selective mineralocorticoid antagonist , such as eplerenone , or low-dose corticosteroids . Currently , abiraterone acetate is being tested in a Phase III trial for men with progressive castration-resistant prostate cancer who are chemotherapy naive . A Phase III trial for patients following prior chemotherapy has been completed and is awaiting analysis . Antitumour activity of docetaxel following treatment with the P05093 inhibitor abiraterone : clinical evidence for cross-resistance ? BACKGROUND : DB05812 and docetaxel are both approved treatments for men with metastatic castration-resistant prostate cancer ( mCRPC ) . DB05812 pre-docetaxel is currently undergoing evaluation in a phase III study . In vitro studies indicate that taxanes may act by disrupting androgen receptor signalling . We hypothesised that prior abiraterone exposure would adversely impact docetaxel efficacy . PATIENTS AND METHODS : We retrospectively evaluated activity of docetaxel in mCRPC patients previously treated with abiraterone , using Prostate Cancer Working Group and radiological criteria . RESULTS : Of the 54 patients treated with abiraterone , 35 subsequently received docetaxel . DB01248 resulted in a prostate-specific antigen ( PSA ) decline of ≥50 % in nine patients [ 26 % , 95 % confidence interval ( CI ) 13 % to 43 % ] , with a median time to PSA progression of 4.6 months ( 95 % CI 4.2 % to 5.9 % ) . PSA declines ≥30 % were achieved by 13 patients ( 37 % , 95 % CI 22 % to 55 % ) . The median overall survival was 12.5 months ( 95 % CI 10.6-19.4 ) . All patients who failed to achieve a PSA fall on abiraterone and were deemed abiraterone-refractory were also docetaxel-refractory ( N = 8 ) . In the 24 patients with radiologically evaluable disease , partial responses were reported in four patients ( 11 % ) , none of whom were abiraterone-refractory . CONCLUSION : The activity of docetaxel post-abiraterone appears lower than anticipated and no responses to docetaxel were observed in abiraterone-refractory patients . P01308 drives transcriptional activity of the P05093 gene in primary cultures of swine theca cells . P01308 stimulates androgen biosynthesis and the accumulation of P05093 mRNA and heterogeneous nuclear ( hn ) RNA in primary cultures of immature swine theca cells . To further assess insulinomimetic transcriptional control , we subcloned 1.007 kilobases ( kb ) of the 5'-upstream region of the P05093 gene ( -976 to +31 base pairs [ bp ] to the transcriptional start site ) into a firefly-luciferase reporter construct . P01308 drove transcriptional activity of this probe in a time- and dose-dependent fashion , with maximal stimulation of 2.7- to 3.2-fold after insulin exposure ( 100 ng/ml ) for 6 h . Progressive deletional constructs -839 , -473 , -174 , and -75/+31 bp delineated expected reduction in responsiveness , except paradoxical gain of basal P05093 promoter activity by the -473/+31-bp sequence . The latter suggests a possible intervening inhibitory sequence . Elimination of all sequences 5'-upstream to -174 bp markedly reduced basal transcriptional activity and abolished insulin action . Point mutation of a presumptive Sp1-like element located within -193/-180 bp inhibited basal and insulin-stimulated luciferase activity of the full-length promoter fragment by 40 % and 67 % , respectively . Disruption of a contiguous presumptive P05549 site produced a comparable outcome . Combined mutation of the Sp1 and P05549 -like elements eliminated basal and insulin-potentiated P05093 promoter activity . By Western analysis , insulin augmented cognate receptor phosphoprotein concentrations by 31-fold within 10 min . Chemical inhibitors of MEK-activated P27361 /2 attenuated insulin-enhanced P05093 transcriptional activity by 76-80 % . In summary , insulin drives transcriptional activity of a 5'-upstream regulatory sequence ( -976 to +31 bp ) of the swine P05093 gene in primary cultures of theca cells , under a minimal requirement for combined activity of proximal ( -193/180 bp ) Sp1 and P05549 -like elements . Synthesis , biological evaluation , and molecular modeling of abiraterone analogues : novel P05093 inhibitors for the treatment of prostate cancer . DB05812 , a steroidal cytochrome P450 17alpha-hydroxylase-17,20-lyase inhibitor ( P05093 ) , is currently undergoing phase II clinical trials as a potential drug for the treatment of androgen-dependent prostate cancer . Since steroidal compounds often show side effects attributable to their structure , we have tried to replace the sterane scaffold by nonsteroidal core structures . The design and synthesis of 20 new abiraterone mimetics are described . Their activities have been tested with recombinant human P05093 expressed in E. coli . Promising compounds were further evaluated for selectivity against P15538 , P19099 , and the hepatic P08684 . Compounds 19 and 20 showed comparable activity to abiraterone ( IC50 values of 144 and 64 nM vs 72 nM ) and similar or even better selectivity against the other CYP enzymes . Selected compounds were also docked into our homology model , and the same binding modes as for abiraterone were found . Changing paradigms in management of metastatic Castration Resistant Prostate Cancer ( mCRPC ) . Recently , the standard of care for metastatic Castration Resistant Prostate Cancer ( mCRPC ) has changed considerably . Persistent androgen receptor ( AR ) signaling has been identified as a target for novel therapies and reengages the fact that AR continues to be the primary target responsible for metastatic prostate cancer . P10275 gene amplification and over expression have been found to result in a higher concentration of androgen receptors on tumor cells , making them extremely sensitive to low levels of circulating androgens . Additionally , prostate cancer cells are able to maintain dihydrotestosterone ( DB02901 ) concentration in excess of serum concentrations to support tumor growth . For many years ketoconazole was the only P05093 inhibitor that was used to treat mCRPC . However , significant toxicities limit its use . Newly approved chemotherapeutic agents such as DB05812 ( an oral selective inhibitor of CYP17A ) , which blocks androgen biosynthesis both within and outside the prostate cancer cells ) , and enzalutamide ( blocks AR signaling ) have improved overall survival . There are also ongoing phase III trials for Orteronel ( P50750 - 700 ) , ARN- 509 and Galeterone ( TOK-001 ) , which targets androgen signaling . In this review , we will present the rationale for the newly approved hormonal treatments , their indications and complications , and we will discuss ongoing trials that are being done to improve the efficacy of the approved agents . Finally , we will talk about the potential upcoming hormonal treatments for mCRPC . DB05812 in heavily pretreated patients with metastatic castrate-resistant prostate cancer . The aim of this study was to evaluate the activity and tolerability of abiraterone acetate in patients with metastatic castrate-resistant prostate cancer treated previously with more than three lines of chemotherapy . Patients received 1 g of abiraterone acetate ( administered as four 250 mg tablets ) orally once daily with prednisone at a dose of 5 mg orally twice daily . The primary endpoint was prostate-specific antigen ( PSA ) response . From August 2011 to January 2013 , 36 patients were enrolled . PSA response was observed in 22 patients ( 61.1 % , 95 % confidence interval : 0.41-0.81 ) . The median time to PSA progression was 7.3 months and after a median follow-up of 10.1 months , all patients were alive . The treatment was generally well tolerated ; side effects secondary to mineralocorticoid excess resulting from blockade of P05093 were largely controlled with prednisone . DB05812 acetate seems to be an effective and well-tolerated treatment option for patients with metastatic castrate-resistant prostate cancer irrespective of the number of chemotherapy lines administered previously . Resistance to abiraterone in castration-resistant prostate cancer : a review of the literature . Persistent androgen signaling is functionally significant in castration-resistant prostate cancer ( CRPC ) and it is actually considered a validated therapeutic target . Residual intra-tumoral androgens compensate for the effects of androgen ablation , activating the androgen receptor ( AR ) , AR-mediated gene expression and driving CRPC . The intra-tumoral biosynthesis of androgens takes place in different ways and cytochrome P450 17A1 ( P05093 ) has a crucial role in this context . DB05812 , a P05093 inhibitor , has shown impressive results in pre- and post-chemotherapy settings , prolonging the survival of patients with CRPC . However , not all patients respond to the treatment and most responders develop resistance , with a widely variable duration of response . Although many hypotheses are emerging , the mechanisms of resistance to abiraterone treatment have not yet been elucidated . The aim of the present review is to describe the main data currently available on resistance to abiraterone . Clinical appraisal of abiraterone in the treatment of metastatic prostatic cancer : patient considerations , novel opportunities , and future directions . While androgen-deprivation therapy can induce dramatic clinical responses in advanced and metastatic prostate cancer , refractory disease ( castration-resistant prostate cancer [ CRPC ] ) eventually emerges . In recent years , several studies have demonstrated the importance of residual intratumoral androgens in maintaining androgen receptor ( AR ) transcriptional activity in CRPC . The cytochrome P450 enzyme P05093 is an obligatory step in androgen synthesis , and therefore a critical therapeutic target in CRPC . DB05812 acetate is a selective , irreversible inhibitor of P05093 and can suppress adrenal synthesis of androgen precursors , and possibly in situ steroidogenesis in the tumor microenvironment . In a phase III multicenter study , abiraterone in combination with prednisone improved median overall survival of men with docetaxel-refractory CRPC by 3.9 months compared to placebo plus prednisone , and also resulted in higher objective prostate-specific antigen and radiographic response rates . The study led to the FDA approval in April 2011 of abiraterone for treatment of chemotherapy-refractory CRPC patients , validating steroidogenesis and the AR axis in general as therapeutic targets in CRPC . The FDA indication for abiraterone was expanded to all CRPCs in December 2012 , while evaluation in even earlier disease states is ongoing . We propose a comprehensive AR axis-targeting approach via simultaneous , frontline enzymatic blockade of several steroidogenic enzymes ( eg , P05093 and P42330 ) in combination with gonadotropin-releasing hormone analogs and potent , second-generation AR antagonists ( eg , enzalutamide ) in order to improve outcomes in patients with prostate cancer . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Beyond castration and chemotherapy : novel approaches to targeting androgen-driven pathways . In castrate-resistant prostate cancer , beyond chemotherapy , existing guidelines suggest only supportive care . However , recent evidence suggests that continued targeting of androgen-dependent pathways may be an efficacious approach . Clinical data is now available for two mechanistically distinct agents ( abiraterone and MDV3100 ) that both ultimately target these pathways . DB05812 is a potent and irreversible inhibitor of P05093 , a critical enzyme in androgen biosynthesis . Phase II studies indicate substantial declines in PSA amongst castrate-resistant patients treated with abiraterone , both prior to and following cytotoxic chemotherapy . In contrast to abiraterone , MDV3100 is a direct inhibitor of the androgen receptor , binding the receptor irreversibly with substantially higher affinity as compared to bicalutamide . A recent phase I/II trial of MDV3100 in castrate-resistant prostate cancer demonstrated tolerability of the agent with activity at the lowest dose level . On the basis of these compelling data , both abiraterone and MDV3100 will be examined in the phase III setting . DB05812 acetate : in metastatic castration-resistant prostate cancer . Oral abiraterone acetate , in combination with prednisone/prednisolone , is used to treat patients with metastatic castration-resistant prostate cancer ( CRPC ) who have previously received docetaxel-containing chemotherapy . DB05812 acetate was developed to specifically inhibit cytochrome P450 (CYP)17A1 , which is an essential enzyme in the biosynthesis of testosterone . In a pivotal phase III trial in patients with metastatic CRPC who have previously received docetaxel-containing chemotherapy , abiraterone acetate 1000 mg once daily plus prednisone 5 mg twice daily significantly prolonged overall survival compared with placebo plus prednisone . In this trial , abiraterone acetate plus prednisone was significantly more effective than placebo plus prednisone in prolonging the time to prostate-specific antigen ( PSA ) progression and in prolonging progression-free survival . Significantly more abiraterone acetate plus prednisone recipients than placebo plus prednisone recipients were considered to be responders , when assessed by PSA levels or radiographic imaging . Treatment with abiraterone acetate plus prednisone in the phase III trial was associated with an acceptable tolerability profile , which was generally similar to that of the placebo plus prednisone group . However , adverse events of special interest ( e.g. cardiac disorders and liver-function test abnormalities and adverse events resulting from elevated mineralocorticoid levels because of P05093 inhibition [ i.e. fluid retention and oedema , hypokalaemia , hypertension ] ) occurred in significantly more abiraterone acetate plus prednisone than in placebo plus prednisone recipients . 17(E)-picolinylidene androstane derivatives as potential inhibitors of prostate cancer cell growth : antiproliferative activity and molecular docking studies . We report a rapid and efficient synthesis of A-ring modified 17α-picolyl and 17(E)-picolinylidene androstane derivatives from dehydroepiandrosterone . Compounds were validated spectroscopically and structurally characterized by X-ray crystallography . Virtual screening by molecular docking against clinical targets of steroidal anticancer drugs ( ERα , AR , P11511 and P05093 ) suggests that 17(E)-picolinylidene , but not 17α-picolyl androstanes could specifically interact with P05093 ( 17α-hydroxylase ) with similar geometry and affinity as DB05812 , a 17-pyridinyl androstane drug clinically used in the treatment of prostate cancer . In addition , several 17(E)-picolinylidene androstanes demonstrated selective antiproliferative activity against PC3 prostate cancer cells , which correlates with DB05812 antiproliferative activity and predicted P05093 binding affinities . Based on these preliminary results , 17(E)-picolinylidene androstane derivatives could be a promising starting point for the development of new compounds for the treatment of prostate cancer . Hormonal therapy for prostate cancer : toward further unraveling of androgen receptor function . Prostate cancer is a major cause of cancer-related death in men . Prostate cancer is an androgen-responsive tumor and the treatment of advanced prostate cancer involves hormonal therapy . First-line treatment for advanced prostate cancer is androgen deprivation therapy ( ADT ) , usually with agents that suppress gonadotropins through a pituitary mechanism . DB00644 agonists and antagonists both suppress gonadal release of testosterone , although their activity profiles vary . ADT down-regulates androgen receptor ( AR ) transcriptional activity in the tumor but the response in metastatic disease is transient and tumors progress as castration-resistant prostate cancer ( CRPC ) . Although serum testosterone concentrations decline dramatically with ADT , CRPC growth remains largely dependent on AR activity . Secondary hormonal therapies are then often employed to further dampen AR-driven transcription . These secondary hormonal therapies either further deplete adrenal or intratumoral androgen synthesis , or directly and competitively antagonize AR . New hormonal agents with both of these mechanisms are in clinical trials and show promising activity in patients with CRPC . DB05812 acetate is an inhibitor of P05093 , which is an enzyme required for the synthesis of all androgens and estrogens . MDV3100 is an AR antagonist that has a higher affinity for AR than any other AR antagonist in clinic use . In phase I and phase II clinical trials , both agents have significant activity . These agents and the promise of the development of others provide hope that more effective hormonal therapies may soon be offered to patients , which will improve clinical outcomes . [ Roles of medical oncologists in the new era of CRPC therapy in Japan ] . Currently , the standard therapy for advanced prostate cancer is endocrine therapy ( luteinizing hormone-releasing hormone [LH-RH]agonists alone or LH-RHagonists plus antiandrogens ) . However , most patients eventually become resistant to these therapies as well as castration therapy . New endocrine therapies for castration-resistant prostate cancer(CRPC)have been developed . DB05812 , a P05093 inhibitor , and enzalutamide , a novel androgen receptor antagonist , have been shown to improve the overall survival , and they are set to be approved in Japan soon . Moreover , docetaxel and cabazitaxel have been established as first- and second-line chemotherapeutic drugs , respectively . Although there is currently no established molecular target drug for CRPC , some drugs such as cabozantinib seem to be effective , and they may be used in the future . In these situations of new drug development , the contribution of medical oncologists is predictable . While medical oncologists can not play central roles in all aspects of drug therapy for urological malignancies in Japan , they must play roles in certain aspects such as new drug development starting from phase I trials , improving multidisciplinary care for adverse events , and promoting translational research . P05093 inhibition as a hormonal strategy for prostate cancer . P10275 ( AR ) signaling has a key role in the pathogenesis of prostate cancer . AR gene amplification , AR overexpression , and activating mutations in the AR occur more frequently as castration-resistant prostate cancer ( CRPC ) evolves , with intratumoral androgen levels remaining sufficient for AR activation despite castration . The source of these androgens might be either adrenal or intratumoral . AR signaling , therefore , remains a valid treatment target for patients with CRPC . P05093 is a key enzyme for androgen biosynthesis . The imidazole antifungal agent ketoconazole weakly and nonspecifically inhibits P05093 , but remains unlicensed for this indication . Chemists at the Cancer Research UK Centre for Cancer Therapeutics have designed a novel , selective , irreversible inhibitor of P05093 called abiraterone , which is more than 20 times more potent than ketoconazole . DB05812 acetate , a prodrug , has undergone phase I assessment , and is rapidly progressing from phase II to phase III trials , in view of its high level of antitumor activity . This agent is safe and well tolerated , and activity profiles suggest that approximately 50 % of CRPC remains AR-ligand driven . Other P05093 inhibitors with alternative mechanisms of action , for example VN/124-1 , are in preclinical development . The rationale for and implications of P05093 inhibition and the P05093 -targeting agents in development are discussed in this Review . DB05812 in prostate cancer : a new angle to an old problem . DB05812 acetate is an orally administered potent inhibitor of cytochrome P450 , family 17 , subfamily A , polypeptide 1 ( P05093 ) , which is essential for synthesis of testosterone from cholesterol . Although decreasing serum testosterone through inhibition of testicular function is the first line of treatment for men with metastatic prostate cancer , residual androgens may still be detected in patients treated with luteinizing hormone-releasing hormone agonists or antagonists . Treatment with abiraterone results in rapid , and complete , inhibition of androgen synthesis in the adrenal glands and potentially within the tumor itself . An overall survival benefit of maximal androgen suppression was recently shown in a randomized placebo-controlled phase III clinical trial of abiraterone with prednisone versus prednisone in men with metastatic castrate-resistant prostate cancer previously treated with docetaxel chemotherapy . DB05812 's efficacy shows the importance of androgen signaling in patients with castrate-resistant metastatic disease , with additional confirmation from recent studies of other novel agents such as MDV3100 , an androgen receptor signaling inhibitor . These promising results now pose a new angle to an old problem about hormonal therapy and raise new questions about how resistance develops , how to best sequence therapy , and how to optimize combinations with other emerging novel agents . [ Mechanisms of resistance to P05093 inhibitors in castrate resistant prostate cancer ] . INTRODUCTION : DB05812 acetate has increased the overall survival of patients with metastatic castration-resistant prostate cancer . However , despite an initial response to treatment , many patients develop resistance to the drug . In this paper we present different hypotheses that may explain the emergence of resistance . METHOD : This review was conducted from the PubMed database . The most relevant articles were selected and analyzed . RESULTS : The molecular mechanisms of resistance to abiraterone acetate remain largely elusive . We detailed some of them including the reactivation of the androgen receptor through alternative biosynthesis of androgens , over expression or mutation of the androgen receptor gene , or the action of co-activators . The over expression of P05093 or the alteration of other genes ' expression involved in steroidogenesis could also contribute to the resistance . CONCLUSION : Some of the molecular mechanisms involved in the resistance to abiraterone acetate were detailed . Better understanding of these mechanisms is a key step to allow the emergence of new therapeutic options and personalized treatments of castration resistant prostate cancer . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . DB05812 inhibits 1α,25-dihydroxyvitamin D3 metabolism by P08684 in human liver and intestine in vitro . The chemopreventive and therapeutic effects of vitamin D3 are exerted through its dihydroxylated metabolite , 1α,25-dihydroxyvitamin D3 [ 1α,25(OH)2D3 ] . Inactivation of 1α,25(OH)2D3 by cytochrome P450 3A4 ( P08684 ) may be an important determinant of its serum and tissue levels . DB05812 , a steroidogenesis inhibitor used in late stage prostate cancer treatment , is a P05093 inhibitor . The purpose of this study was to assess the potential of abiraterone to block hepatic and intestinal inactivation of biologically active vitamin D3in vitro and to evaluate if abiraterone can alter P08684 marker substrate activities . Biotransformation reactions were initiated with NADPH regenerating solutions following initial preincubation of pooled human hepatic or intestinal microsomal protein or human recombinant P08684 supersomes with 1α,25(OH)2D3 , midazolam or triazolam for 10min at 37°C . Formation of hydroxylated metabolites of 1α,25(OH)2D3 , midazolam or triazolam was analyzed by liquid chromatography-mass spectrometry method . Co-incubation of 1α,25(OH)2D3 with abiraterone at varying concentrations ( 0.2-100μM ) led to up to ∼85 % inhibition of formation of hydroxylated metabolites of 1α,25(OH)2D3 thus preventing inactivation of active vitamin D3 . The IC50 values for individual metabolites of 1α,25(OH)2D3 ranged from 0.4 to 2.2μM in human liver microsomes or human intestinal microsomes . The mechanism of P08684 -mediated inhibition of 1α,25(OH)2D3 by abiraterone was competitive ( apparent Ki 2.8-4.3μM ) . Similar inhibitory effects were also observed upon inclusion of abiraterone into midazolam or triazolam hydroxylation assays . In summary , our results suggest that abiraterone inhibits the P08684 -mediated inactivation of active vitamin D3 in human liver and intestine , potentially providing additional anti-cancer benefits to prostate cancer patients . This article is part of a Special Issue entitled ' 16th Vitamin D Workshop ' . [ Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture ] . In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10(-7)-10(-10) M it protected cells of sympathetic ganglia , neocortex and continues cell lines under damaging actions of H2O2 ( 0.0001 M ) , NH4CI ( 0.01 M ) and cooling . DB00086 essentially influenced the mode of damaging effect of DB00171 ( 0.001 M ) . Even a short-term exposition ( 20 min ) of PC12 cells with both proteins ( each in the concentration 10(-9) M ) led to sharp alterations in intracellular DB00171 - or Ca(2+)-activated proteolysis . In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation , improvement of cell adhesion , high survival rate , the increase in quantity and length of processes and their arborisation . Electronic microscopy established the character of structural rearrangements of nervous tissue cells ( neurons , astrocytes , oligodendrocytes ) , reflecting the protective action of plasminogen and streptokinase . In the presence of plasminogen and especially streptokinase , the total number of cultured glioma P13671 and neuroblastoma IMR-32 cells , the intracellular contents of protein , RNA and DNA increased several-fold . Addition of plasminogen promoted formation of processes by neuroblastoma cells , this suggests initiation of differentiation of cellular elements . In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells . These proteins did not cause transformation of PC12 enterochromaffine cells to neurons , though plasminogen facilitated it . P00747 addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium , and streptokinase did not lose its plasminogen-activating capacity . Synthesis , biological evaluation , and molecular modeling studies of methylene imidazole substituted biaryls as inhibitors of human 17alpha-hydroxylase-17,20-lyase ( P05093 ) -- part II : Core rigidification and influence of substituents at the methylene bridge . Thirty-five novel substituted imidazolyl methylene biphenyls have been synthesized as P05093 inhibitors for the potential treatment of prostate cancer . Their activities have been tested with recombinant human P05093 expressed in Escherichia coli . Promising compounds were tested for selectivity against P15538 , P19099 , and hepatic CYP enzymes 3A4 , 1A2 , 2B6 and 2D6 . The core rigidified compounds ( 30-35 ) were the most active ones , being much more potent than Ketoconazole and reaching the activity of DB05812 . However , they were not very selective . Another rather potent and more selective inhibitor ( compound 23 , IC(50)=345 nM ) was further examined in rats regarding plasma testosterone levels and pharmacokinetic properties . Compared to the reference DB05812 , 23 was more active in vivo , showed a longer plasma half-life ( 10h ) and a higher bioavailability . Using our P05093 homology protein model , docking studies with selected compounds were performed to study possible interactions between inhibitors and amino acid residues of the active site .

MH_train_1003

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MH_train_1003

interacts_with DB00790?

multiple_choice

A 3-D model for P08908 -receptor agonists based on stereoselective methyl-substituted and conformationally restricted analogues of 8-hydroxy-2-(dipropylamino)tetralin . The enantiomers of cis- and trans-1,2,3,4,4a,5,10,10a-octahydro-9-hydroxy-1- propylbenzo[g]quinolines ( 10 and 11 , respectively ) and the enantiomers of trans-1,2,3,4,4a,5,6,10b-octahydro-10- hydroxy-4-propylbenzo[f]quinoline ( 12 ) have been synthesized and their stereochemical and conformational characteristics have been studied by use of X-ray crystallography and molecular mechanics ( P08253 ) calculations . The compounds , which are conformationally restricted analogues of the potent 5-hydroxytryptamine ( 5-HT ) receptor agonist 8-hydroxy-2- (dipropylamino)tetralin ( 8-OH-DPAT ; 1 ) have been evaluated for central 5-HT and dopamine receptor stimulating activity by use of biochemical and behavioral tests in rats . In addition , we have evaluated the ability of these compounds and a number of previously reported analogues to displace [ 3H ] -8-OH-DPAT from P08908 -binding sites . The enantiomers of 12 behave as potent P08908 -receptor agonists , whereas the octahydrobenzo[g]quinoline derivatives are much less potent or inactive . In general , the affinities of the compounds correlate well with their agonist potencies . The set of compounds under study is accommodated by a novel computer-graphics-derived model for P08908 -receptor agonism . The model consists of a flexible pharmacophore and a partial receptor-excluded volume . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Cardiac protective effect of Astragalus on viral myocarditis mice : comparison with DB00790 . In clinical practice , Astragali Radix ( Astragalus ) , the root of Astragalus membranaceus Bunge , has been widely applied to treat patients with viral diseases , including viral myocarditis in China . The present study was designed to evaluate the protective effects of Astragalus on the function of sarcoplasmic reticulum calcium ATPase ( P16615 ) activity and endothelin system at acute and chronic periods of myocarditis mice induced by CVB(3) infection . Astragalus feeding ( 2.2 mg/kg/day ) could significantly increase the survival rate , alleviate pathological alterations and serum cardiac troponin I ( cTnI ) , as well as restore impaired SERCA activity at the acute stage . Low affinity and capacity of ETR were reversed with Astragalus after the first CVB(3) inoculation up to 7 days and after the second virus inoculation up to 150 days . In the meantime , the contents of cardiac ET-1 and P01160 were reduced . Comparison the myocarditis mice treated with DB00790 ( 0.44 mg/kg/day ) , an P12821 inhibitor , shows that Astragalus achieved a similar effect on survival rate , P16615 and ET system . These results indicated that the beneficial effects of Astragalus and DB00790 for treating viral myocarditis might be partly mediated by preserving the functions of SERCA 2 activity and ET system . [ DB00988 -beta-hydroxylaseaktivität im Plasma von Dialysepatienten ( author 's transl ) ] . Plasma dopamin-b-hydroxylase ( P09172 ) was studied in 70 healthy control persons and in 37 hemodialysed patients . Basal P09172 in controls corresponded to 50.0 +/- 29.3 IU . There was was no significant difference between males ( 53.9 +/1 33.8 IU ) and females ( 47.4 +/- 25 IU ) ; no correlation could be found between age and plasma P09172 . In hemodialysed patients basal P09172 levels were significantly ( p less than 0.01 ) decreased ( 32.5 % /- 17.6 IU ) , suggesting lowered sympathetic activity and/or abnormalities in release , distribution space , or metabolism of P09172 . During hemodialysis plasma P09172 activity rose during ultrafiltration . This finding indicates a directionally appropriate sympathetic reflex response to volume depletion in dialysed patients . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . Expression profile of genes associated with the dopamine pathway in vitiligo skin biopsies and blood sera . BACKGROUND : Dopamine has been proven to be toxic for melanocytes . In vitiligo patients the level of dopamine is increased and the functioning of several enzymes participating in the dopamine pathway is changed . METHODS : With the use of quantitative real-time polymerase chain reaction and ELISA the expression of genes connected to the dopamine pathway ( PAH , P61457 , TH , DDC , P09172 , PNMT , P07203 , P21397 , P27338 , P21964 , P21728 - P21918 , P54219 and Q05940 ) was observed in vitiligo patients ' and control subjects ' skin and blood . RESULTS : The mRNA expression of P07203 , DDC , P21397 , P21728 and P21918 differs in vitiligo skin and the protein level of DDC , P21397 , P27338 , P21728 and P21918 is changed in vitiligo patients ' skin and/or blood sera . CONCLUSIONS : The dopamine pathway probably influences melanogenesis directly or through the melanocortin pathway . We provide new data about changes of expression profile of the dopamine-synthesizing enzyme DDC , the dopamine-degrading enzymes P21397 and P27338 and the D1-like family dopamine receptors in vitiligo skin and blood sera . Bayesian meta-analysis of tissue angiotensin-converting enzyme inhibitors for reduction of adverse cardiovascular events in patients with diabetes mellitus and preserved left ventricular function . The role of angiotensin-converting enzyme ( P12821 ) inhibitors in diabetic patients with preserved ventricular function is uncertain . Tissue P12821 inhibitors have been defined by increased lipophilicity and structural characteristics that result in greater tissue-specific P12821 binding when compared with plasma P12821 inhibitors . A Bayesian meta-analysis of randomized trials was conducted to evaluate tissue P12821 inhibitors in prevention of cardiovascular disease among patients with diabetes mellitus and preserved left ventricular function . Four trials were selected that evaluated 2 different P12821 inhibitors and included 10,328 patients ( 43,517 patient-years ) . The DB00790 Substudy in Coronary Artery Disease and Diabetes ( PERSUADE ) and the DB00790 Protection Against Recurrent Stroke Study ( PROGRESS ) compared the effects of perindopril vs a placebo , and the Heart Outcomes Prevention Evaluation ( HOPE ) and the Non- P01308 -Dependent Diabetes , Hypertension , Microalbuminuria , Proteinuria , Cardiovascular Events , and Ramipril ( DIABHYCAR ) study investigated the impact of ramipril vs a placebo . Bayesian meta-analysis of sequential trials and sensitivity analysis of therapeutic response were subsequently computed . Bayesian meta-analysis determined reduced risk of cardiovascular mortality ( PB=.991 ) , myocardial infarction ( PB=.999 ) , and the need for invasive coronary revascularization ( PB=.995 ) when compared with placebo . Total mortality was also decreased ( PB=.967 ) , while the risk of stroke ( PB=.907 ) and hospitalization for heart failure ( PB=.923 ) were impacted . Bayesian meta-analysis of randomized trials suggests that tissue P12821 inhibitors decrease the probability that diabetic patients with preserved left ventricular function will experience myocardial infarctions and cardiovascular death and reduce overall mortality . Fetal corticotrophin-releasing hormone mRNA , but not phosphatidylserine-exposing microparticles , in maternal plasma are associated with factor VII activity in pre-eclampsia . BACKGROUND : Pre-eclampsia is associated with increased placental debris circulating in maternal plasma . OBJECTIVES : This study related placental debris to maternal markers of coagulation and endothelial activation in pre-eclampsia . PATIENTS/METHODS : Circulating fetal corticotrophin-releasing hormone ( P06850 ) mRNA and phosphatidylserine ( PS ) -exposing microparticles were assayed in third trimester plasma from women with pre-eclampsia ( n = 32 ) and controls ( n = 32 ) matched for age , body mass index , parity , and gestational age at sampling . Markers of maternal hemostasis and endothelial function were assessed . RESULTS : Fetal P06850 mRNA levels were higher in pre-eclampsia [ mean 0.75 ( SD 2.77 ) P06850 /glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) mRNA ratio ] than in control pregnancies [ 0.20 ( 0.74 ) , P = 0.014 ] . PS-exposing microparticle levels were not different between the groups . Women with pre-eclampsia had higher levels of tissue factor pathway inhibitor ( P10646 ) , prothrombin F(1+2) fragment ( F(1+2) ) , factor XIIa , soluble vascular cell adhesion molecule 1 , P04275 and plasminogen activator inhibitor 1 than controls . Fetal P06850 mRNA correlated with P10646 in pre-eclampsia and control groups ( r = 0.38 , P = 0.031 , and r = 0.37 , P = 0.039 , respectively ) . Fetal P06850 mRNA correlated with FVII activity ( r = 0.43 , P = 0.017 ) and PS-exposing microparticles correlated inversely with F(1+2) ( r = -0.64 , P < 0.001 ) in pre-eclampsia . CONCLUSIONS : Placental debris , assessed by fetal P06850 mRNA levels in maternal blood , is related to coagulation potential , i.e. FVII activity , but not to markers of coagulation or endothelial activation in pre-eclampsia . Atrial natriuretic peptide : a possible mediator involved in dexamethasone 's inhibition of cell proliferation in multiple myeloma . Atrial natriuretic peptide ( P01160 ) has been recognized for several decades for its role of regulating blood pressure . Recently , cumulating evidences show that P01160 plays an anticancer role in various solid tumors via blocking the kinase cascade of Ras- Q02750 /2- P27361 /2 with the result of inhibition of DNA synthesis . P01160 , as well as its receptors ( P16066 and P17342 ) has been identified present in the embryonic stem cell and a wide range of cancer cells . Various lymphoid organs , such as lymph nodes , have been detected the presence of P01160 . Multiple myeloma ( MM ) , though the therapies have evolved significantly , is still an incurable disease as B lymphocyte cell neoplasm . Dexamethasone is the cornerstone in treatment of MM via inactivation of Ras- Q02750 /2- P27361 /2 cascade reaction . Coincidently , dexamethasone can increase the expression of P01160 markedly . Nevertheless , the role of P01160 in MM is unclear . Based on these results above , we raise the hypothesis that P01160 is involved in mediating dexamethasone 's inhibition of proliferation in MM cells , which suggests that P01160 may be a potential agent to treat MM . Sexually dimorphic stress and pro-inflammatory cytokine responses to an intravenous corticotropin-releasing hormone challenge of Brahman cattle following transportation . This study was designed to characterize potential sexually dimorphic stress and immunological responses following a corticotropin-releasing hormone ( P06850 ) challenge in beef cattle . Six female ( heifers ) and six male ( bulls ) Brahman calves ( 264 ± 12 d of age ) were administered P06850 intravenously ( 0.5 µg of P06850 /kg body mass ) after which serum concentrations of cortisol increased from 0.5 h to 4 h . From 1 h to 4 h after P06850 administration , serum cortisol concentrations were greater in heifers than in bulls . In all cattle , increased serum concentrations of P01375 -α , P05231 and IFN-γ were observed from 2.5 h to 3 h after P06850 , with greater concentrations of IFN-γ and P05231 in heifers than bulls . Heifer total leukocyte counts decreased 1 h after P06850 administration , while bull leukocyte counts and percent neutrophils decreased 2 h after P06850 administration . Heifers had greater rectal temperatures than bulls , yet rectal temperatures did not change following administration of P06850 . There was no effect of P06850 administration on heart rate . However , bulls tended to have increased heart rate 2 h after P06850 administration than before P06850 . Heifer heart rate was greater than bulls throughout the study . These data demonstrate that acute P06850 administration can elicit a pro-inflammatory response , and cattle exhibit a sexually dimorphic pro-inflammatory cytokine and cortisol response to acute P06850 administration . DB00819 inhibits aquaporin-1 expression and colon cancer xenograft tumor growth . BACKGROUND/AIMS : To study the effects of water channel protein inhibitor acetazolamide on xenograft tumor growth of colon cancer in nude mice . METHODOLOGY : Setting up human colon cancer model in nude mice , mice were randomly divided into two groups as experimental group and control group . DB00819 was given at a volume of 0.1mL per mice ( 40mg/kg/d , ig ) in experimental group , while the same volume of sterile saline was given in control group ( ig ) . After 21 days , protein and m-RNA levels of P29972 in tumor tissues from two groups were detected respectively by Western blot and RT-PCR to evaluate the treatment effects . P29972 , P15692 and P28906 expression was detected by immunohistochemistry , simultaneously . RESULTS : DB00819 ( 40mg/kg/d , ig ) significantly inhibited the xenograft tumor growth of colon cancer in nude mice . The inhibition rate was 88.28 % . In comparison with the control group , P29972 protein and mRNA level were significantly reduced in the experimental group ( p < 0.01 ) . P29972 , P15692 and P28906 expression in experimental group were positively correlated between each other ( p < 0.01 ) . CONCLUSIONS : DB00819 can suppress the xenograft tumor growth by inhibiting the expression of P29972 . The role of tumor necrosis factor-alpha in Henoch-Schonlein purpura . Henoch-Schonlein purpura ( HSP ) is one of the most common types of vasculitis disorders in childhood and is characterized by a rash , arthritis , abdominal pain , and renal involvement . The factors that determine and mediate the severity of HSP and its renal involvement remain poorly understood , although it is likely that pro-inflammatory cytokines , including tumor necrosis factor-alpha ( P01375 ) , are involved in the pathogenesis . Serum and urine levels of P01375 were measured in children with HSP in the acute and convalescent phases by ELISA . Serum P01375 levels were significantly higher in proteinuric HSP in the acute phase ( 36.6+/-8.5 pg/ml ) compared with those with HSP without renal involvement and those with hematuric HSP ( 25.4+/-4.5 and 27.1+/-3.9 pg/ml ) ( P < 0.005 ) . However , these significantly higher levels disappeared in the convalescent phase . Using matched serum samples from the same patients , serum P01375 levels of proteinuric HSP patients were significantly lower in the convalescent phase ( 29.9+/-4.6 pg/ml , P < 0.05 ) than in the acute phase ( Q04695 +/-8.2 pg/ml ) . Although urine P01375 levels were higher in proteinuric HSP in the acute phase and reduced in the convalescent phase , there were no significantly high or low levels . These results suggest that increased P01375 levels in the serum induce a series of functional and morphological changes in the glomerular cells in the acute phase and may be used as markers for monitoring the disease activity of HSP with severe renal involvement . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . Quantitative and qualitative pleiotropic differences between Simvastatin single and Vytorin combination therapy in hypercholesterolemic subjects . AIMS : This cross-sectional study tested the hypothesis that treatment with the combination of DB00973 /Simvastatin ( Vytorin ) leads to broader changes in the expression levels of immunomodulatory genes as compared to Simvastatin monotherapy . METHODS : Illumina 's GenomeStudio gene expression module was used to compare gene profiles of Vytorin and Simvastatin in the peripheral blood mononuclear cells of 20 hypercholesterolemic subjects . RESULTS : The characteristics of the immunomodulatory genes , which were altered by Vytorin , differed from those genes which were altered by Simvastatin . Vytorin mostly altered the expression levels of genes related to inflammation/oxidative stress ; it downregulated the NF-KappaB and upregulated the expression of anti-inflammatory cytokine , P22301 , and anti-oxidant enzymes , P07203 and P04179 , but also upregulated the expression levels of genes involved in cellular activation , adhesion , and coagulation cascade , including P04275 , P08709 , P02776 , P10720 SELP , P05106 , P18084 . Simvastatin mostly altered the expression levels of genes related to cellular apoptosis/proliferation . It upregulated the expression levels of apoptosis-related genes O14727 , Q07812 , P46695 , and P07333 , and downregulated the expression levels of genes related to cellular proliferation , including P21246 and Q07108 . Treatment with Vytorin combination therapy modulated lipid profile and serum levels of the P02741 more effectively , than treatment with Simvastatin monotherapy . CONCLUSION : The nature of the pleiotropic effects may play a role in Vytorin 's and Simvastatin 's clinical efficacies . Different effects of perindopril and enalapril on monocyte cytokine release in coronary artery disease patients with normal blood pressure . BACKGROUND : Favorable effects of angiotensin-converting enzyme ( P12821 ) inhibitor treatment on the incidence of cardiovascular and cerebrovascular mortality and morbidity are not limited to patients with elevated blood pressure . As suggested by our previous results , the physicochemical and pharmacokinetic differences between drugs may markedly contribute to the strength of pleiotropic effects of P12821 inhibitors . METHODS : The present study was aimed at comparing the effects of serum- and tissue-type P12821 inhibitors on monocyte release of proinflammatory cytokines in normotensive patients with stable coronary artery disease . The participants were randomized to 90-day treatment with enalapril ( 20 mg daily , n = 29 ) , perindopril ( 4 mg daily , n = 27 ) or placebo ( n = 28 ) . Plasma levels of lipids , glucose , insulin and high sensitivity P02741 ( hsCRP ) , as well as monocyte release of proinflammatory cytokines were determined before and after 30 days of therapy , and at the end of the treatment . RESULTS : Lipopolysaccharide-stimulated monocytes from normotensive patients with stable coronary artery disease released significantly more P01375 -α , interleukin-1β and monocyte chemoattractant protein-1 in comparison with monocytes from 23 matched control subjects . Their baseline hsCRP levels were also higher . DB00790 reversed the disease-induced changes in cytokine release and reduced plasma hsCRP , while the effect of enalapril was much more limited . The effect on both drugs on cytokine release was stronger in insulin-resistant than insulin-sensitive subjects . CONCLUSIONS : Our results indicate that perindopril is superior to enalapril in producing monocyte-suppressing and systemic anti-inflammatory effects in normotensive patients with coronary artery disease . This action may contribute to the clinical effectiveness of tissue P12821 inhibitors in the therapy of atherosclerosis-related disorders , particularly in insulin-resistant subjects . Comparing interfertility data with random amplified microsatellites DNA ( RAMS ) studies in Ganoderma Karst. Taxonomy . The taxonomy of the causal pathogen of basal stem rot of oil palms , Ganoderma is somewhat problematic at present . In order to determine the genetic distance relationship between G. boninense isolates and non-boninense isolates , a random amplified microsatellites DNA ( RAMS ) technique was carried out . The result was then compared with interfertility data of G. boninense that had been determined in previous mating studies to confirm the species of G. boninense . Dendrogram from cluster analysis based on UPGMA of RAMS data showed that two major clusters , I and II which separated at a genetic distance of 0.7935 were generated . Cluster I consisted of all the biological species G. boninense isolates namely CNLB , GSDK 3 , O15534 71 , WD 814 , Q9BVC4 3 , Q9BVC4 6 , OC , GH 02 , 170 SL and 348781 while all non-boninense isolates namely G . Q9H6B4 , WRR , TFRI 129 , G . RES , GJ , and CNLM were grouped together in cluster II . Although the RAMS markers showed polymorphisms in all the isolates tested , the results obtained were in agreement with the interfertility data . Therefore , the RAMS data could support the interfertility data for the identification of Ganoderma isolates . Prolonged oxytocin treatment in rats affects intracellular signaling and induces myocardial protection against infarction . DB00107 is a hormone , which is released into the circulation in response to acute or chronic stress stimuli . One of the important targets of oxytocin is cardiovascular system . Present studies were aimed at testing the hypothesis that prolonged treatment with oxytocin ( simulation of stress-induced rise in circulating oxytocin ) activates intracellular signaling pathways playing a role in ischemia/reperfusion injury . Furthermore , we tested protective effects of oxytocin treatment in vivo against cardiac injury induced by ischemia/reperfusion of isolated hearts . Male Wistar rats were treated with oxytocin or vehicle continuously via osmotic minipumps for 2 weeks . The hearts were used for biochemical measurements or isolated for Langendorff perfusion . Treatment with oxytocin resulted in a significant increase in specific phosphorylation ( activation ) of p38-MAPK and Akt kinase , an increase in phosphorylated Hsp27 and an elevation in atrial natriuretic peptide ( P01160 ) levels in left ventricular heart tissue . There were no significant changes in the activation of P08253 and P29323 in the left heart ventricle of oxytocin-treated rats . Postischemic recovery of functional parameters LVDP , RPP , +dP/dtmax and -dP/dtmax was better in the hearts of oxytocin-treated rats compared to that in the controls . DB00107 treatment significantly reduced infarct size to 15.1 + 3.2 % as compared to 32.4 + 3.5 % in vehicle-treated rats ( p < 0.01 ) . This is the first evidence for cardioprotective effects of oxytocin administered in vivo simulating chronic stress-induced elevation in plasma oxytocin . The present results show that positive effects of oxytocin that may ameliorate negative consequences of stress on the heart are , at least in part , mediated through p38-MAPK and Akt kinase pathways . Inhibition of the renin-angiotensin system improves physiological outcomes in mice with mild or severe cancer cachexia . Cancer cachexia describes the progressive skeletal muscle wasting and weakness associated with many cancers . Cachexia reduces mobility and quality of life and accounts for 20-30 % of all cancer-related deaths . Activation of the renin-angiotensin system causes skeletal muscle wasting and weakness . We tested the hypothesis that treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , perindopril , would enhance whole body and skeletal muscle function in cachectic mice bearing Colon-26 ( C-26 ) tumors . CD2F1 mice received a subcutaneous injection of phosphate buffered saline or C-26 tumor cells inducing either a mild or severe cachexia . The following day , one cohort of C-26 mice began receiving perindopril in their drinking water ( 4 mg kg(-1) day(-1) ) for 21 days . In mild and severe cachexia , perindopril increased measures of whole body function ( grip strength and rotarod ) and reduced fatigue in isolated contracting diaphragm muscle strips ( p < 0.05 ) . In severely cachectic mice , perindopril reduced tumor growth , improved locomotor activity and reduced fatigue of tibialis anterior muscles in situ ( p < 0.05 ) , which was associated with increased oxidative enzyme capacity ( succinate deyhydrogenase , p < 0.05 ) . DB00790 attenuated the increase in Q969Q1 and P05231 mRNA expression and enhanced Akt phosphorylation in severely cachectic mice but neither body nor muscle mass was increased . These findings support the therapeutic potential of P12821 inhibition for enhancing whole body function and reducing fatigue of respiratory muscles in early and late stage cancer cachexia and should be confirmed in future clinical trials . Since P12821 inhibition alone did not enhance body or muscle mass , co-treatment with an anabolic agent may be required to address these aspects of cancer cachexia . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Q96RP3 is expressed in human pregnant myometrial cells and regulates myosin light chain phosphorylation : potential role of the type-2 corticotropin-releasing hormone receptor in the control of myometrial contractility . The family of P06850 -related peptides are suggested to play important roles in the control of myometrial contractility during pregnancy and labor . In this study we investigated the expression of urocortin II ( P55089 II ) in human myometrium and its ability to phosphorylate intracellular components that can be involved in modulating myometrial contractility . Using RT-PCR and fluorescent in situ hybridization , we demonstrated that P55089 II and type-2 P06850 receptor ( Q13324 ) mRNAs were expressed in human nonpregnant and pregnant myometrium . Immunofluorescent studies confirmed protein expression of P55089 II in human pregnant myometrial cells , whereas chemical cross-linking studies with radiolabeled P55089 II confirmed the presence of Q13324 sites with an apparent molecular mass of 50 kDa . Treatment of primary human myometrial cells with P55089 II to specifically activate Q13324 resulted in a dose-dependent increase of myosin light chain ( MLC(20) ) phosphorylation . Activation of protein kinase C ( PKC ) and P27361 /2 was required for the P55089 II-induced activation of MLC(20) , because treatment of myometrial cells with inhibitors of MAPK kinase 1 ( U0126 ) and PKC ( bisindolylmaleimide ) inhibited the P55089 II-induced phosphorylation of MLC(20) . Furthermore , the P55089 II effect on MLC(20) was dependent on RhoA translocation to the membrane and subsequent activation of RhoA-associated kinase , as shown by the use of the specific inhibitors exoenzyme P01024 and Y27632 . Collectively , our data suggest a distinctive role for Q13324 - specific agonists like P55089 II in the control of myometrial contractility during human pregnancy involving sequential activation of PKC , MAPK kinase 1 , P27361 /2 , RhoA , and RhoA-associated kinase , leading to the MLC(20) phosphorylation . DB00203 induces angiogenic response in human coronary arteriolar endothelial cells through the expression of thioredoxin , hemeoxygenase and vascular endothelial growth factor . This study was undertaken to investigate the effect of phosphodiesterase-5 ( O76074 ) inhibitor , sildenafil , on angiogenic response in human coronary arteriolar endothelial cells ( HCAEC ) . The cells exposed to sildenafil ( 1-20 microM ) demonstrated significantly accelerated tubular morphogenesis with the induction of thioredoxin-1 ( P10599 -1 ) , hemeoxygenase-1 ( P09601 ) and P15692 . DB00203 induced P15692 and angiopoietin specific receptors such as P35968 , Tie-1 and Tie-2 . This angiogenic response was repressed by tinprotoporphyrin IX ( SnPP ) , an inhibitor of P09601 enzyme activity . DB00203 below 1 muM has no angiogenic effect as evidenced by reduced tuborogenesis . DB00203 along with SnPP inhibited both P15692 and Q15389 ( Ang-1 ) protein expression . Therefore our results demonstrated for the first time that sildenafil is a very potent pro-angiogenic factor . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . The use of microcalorimetry and HPLC for the determination of degradation kinetics and thermodynamic parameters of DB00790 Erbumine in aqueous solutions . DB00790 Erbumine ( O15534 ) is one of the newly used angiotensin-converting enzyme inhibitors ( P12821 inhibitors ) and is used for the treatment of patients with hypertension and symptomatic heart failure . It has two main degradation pathways , i.e. the degradation by hydrolysis and the degradation by cyclization . An isothermal heat conduction microcalorimetry ( MC ) and high pressure liquid chromatography ( HPLC ) were used for the characterization of aqueous solutions of O15534 and its stability properties . The rates of heat evolved during degradation of perindopril were measured by MC as a function of temperature and pH and from these data rate constant and change in enthalpy of the reactions were determined . With the HPLC method the concentration of perindopril and its degradation products were measured as a function of time in aqueous solutions of different pH that were stored at different temperatures . We demonstrated that reactions of degradation of perindopril at observed conditions follow the first order kinetics . The Arrhenius equation for each pH was determined . At pH 6.8 only one degradation pathway is present , i.e. the degradation by hydrolysis . Degradation constants for this pathway calculated from MC data are in good agreement with those obtained from HPLC . MC as a non-specific technique was shown to be useful in studies of O15534 when one reaction was present in the sample and also when more chemical and physical processes were simultaneously running . Investigation of mechanisms mediating 8-OH-DPAT-induced impairment of spatial memory : involvement of P08908 receptors in the dorsal hippocampus in rats . The purpose of this study was to identify mechanisms that mediate the impairment of spatial memory induced by 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 / P34969 receptor agonist , in the eight-arm radial maze in rats . WAY-100635 and NAN-190 , P08908 receptor antagonists , reversed the impairment of spatial memory induced by systemic injection of 8-OH-DPAT ( 1 mg/kg , i.p. ) . On the other hand , the alpha1-adrenoceptor antagonist prazosin and a selective P34969 receptor antagonist SB269970 had no effect on 8-OH-DPAT-induced impairment of spatial memory . Bilateral microinjection of 8-OH-DPAT ( 4 microg/side ) impaired spatial memory when injected into the dorsal hippocampus ( DH ) . Contrastingly , spatial memory was unaffected by microinjections of 8-OH-DPAT into the other six areas examined : ventral hippocampus ( VH ) , central amygdaloid nucleus ( P12821 ) , lateral hypothalamus ( LH ) , nucleus accumbens ( NAc ) , and dorsal ( DR ) and median ( MR ) raphe nucleus . Furthermore , NAN-190 significantly reversed the impairment of spatial memory induced by intra-DH injection of 8-OH-DPAT . These findings suggest that P08908 receptors in the DH play an important role in the mechanisms underlying the 8-OH-DPAT-induced impairment of spatial memory in rats . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Comparison of effects of clodronate and zoledronic acid on the repair of maxilla surgical wounds - histomorphometric , receptor activator of nuclear factor-kB ligand , osteoprotegerin , P04275 , and caspase-3 evaluation . BACKGROUND : The aim of this study was to compare clodronate and zoledronic acid regarding their influence on the repair of surgical wounds in maxillae ( soft tissue wound and tooth extraction ) and their relation to osteonecrosis . MATERIAL AND METHODS : Thirty-four Wistar rats were allocated into three groups according to the treatment received : ( i ) 12 animals treated with zoledronic acid , ( ii ) 12 animals treated with clodronate and ( iii ) 10 animals that were given saline solution . All animals were subjected to tooth extractions and surgically induced soft tissue injury . Histological analysis of the wound sites was performed by means of hematoxylin-eosin ( H & E ) staining and immunohistochemical staining for receptor activator of nuclear factor-kB ligand ( O14788 ) , osteoprotegerin ( O00300 ) , P04275 , and caspase-3 . RESULTS : The zoledronic acid group showed higher incidence of non-vital bone than did the clodronate group at the tooth extraction site . At the soft tissue wound site , there were no significant differences in non-vital bone between the test groups . O14788 , O00300 , P04275 , and caspase-3 did not show significant differences between the groups for both sites of surgical procedures . CONCLUSION : Both of the bisphosphonates zoledronic acid and clodronate are capable of inducing maxillary osteonecrosis . Immunohistochemical analysis suggests that the involvement of soft tissues as the initiator of osteonecrosis development is less probable than has been pointed out . P12821 activity is involved in the mechanism of increased endogenous nitric oxide synthase inhibitor in patients with type 2 diabetes mellitus . The renin-angiotensin system plays an important role in the elevation of asymmetric dimethylarginine ( DB01686 ) , an endogenous inhibitor of nitric oxide synthase , in hypertensive patients , so the present study was designed to examine whether angiotensin-converting enzyme ( P12821 ) activity is also involved in the mechanism of DB01686 elevation in type 2 diabetes mellitus ( NIDDM ) . A crossover study was performed to determine if P12821 inhibition with perindopril ( 4 mg/day ) for 4 weeks decreases serum DB01686 concentration and plasma P04275 ( P04275 ) level ( a marker of endothelial injury ) in 11 patients with NIDDM . None of the patients was treated with insulin or oral hypoglycemic drugs , and none had major diabetic complications . Before the protocol began , serum DB01686 and plasma P04275 were significantly higher in the 11 NIDDM patients , when compared with 8 control subjects without diabetes . DB00790 did not affect blood pressure or glucose metabolism , but did significantly decrease serum DB01686 and plasma P04275 . These results suggest that endothelial injury associated with DB01686 elevation may be present even in patients with non-complicated NIDDM , and that increased activity of P12821 may be involved in such endothelial dysfunction . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . P00797 -angiotensin system is involved in the mechanism of increased serum asymmetric dimethylarginine in essential hypertension . Endothelium-dependent/nitric oxide ( NO ) -mediated vasodilation is impaired in hypertensive individuals . DB01686 ( DB01686 ) , an endogenous inhibitor of NO synthase , is synthesized by many types of cells including vascular endothelial cells . The serum level of DB01686 is elevated in patients with essential hypertension , but the mechanism for this increase is unknown . Therefore , the present study examined whether the renin-angiotensin system ( DB01367 ) is involved . Patients with essential hypertension [ systolic blood pressure ( BP ) > 160 mmHg and/or diastolic BP > 95 mmHg ] were randomized to an angiotensin-converting enzyme ( P12821 ) inhibitor treatment group ( perindopril , 4mg/day for 4 weeks , n = 7 ) , an angiotensin II type 1 ( AT1 ) receptor antagonist treatment group ( losartan , 50 mg/day for 4 weeks , n = 7 ) or a beta-blocker treatment group ( bisoprolol , 5 mg/day for 4 weeks , n = 7 ) . Before and after the treatment , BP , serum concentration of DB01686 and plasma concentration of P04275 ( P04275 , a biological marker of endothelial injury ) were measured . DB00790 , losartan and bisoprolol decreased BP to a similar extent , and either perindopril or losartan , but not bisoprolol , significantly decreased serum DB01686 and plasma P04275 . These findings suggest that the DB01367 may contribute to the mechanism of increased serum DB01686 as well as to the endothelial injury observed in hypertensive patients . The vasculoprotective actions of P12821 inhibitors or AT1 receptor antagonists may be explained at least in part by amelioration of the endothelial injury through a decrease in the serum DB01686 concentration . Vascular endothelial growth factor expression and glomerular endothelial cell loss in the remnant kidney model . BACKGROUND : Vascular endothelial growth factor ( P15692 ) is constitutively expressed in the glomerulus where it may have a role in the maintenance of capillary endothelial cell integrity . The present study sought to examine changes in P15692 expression in a model of progressive renal disease and to assess the effects of angiotensin converting enzyme ( P12821 ) inhibition . METHODS : Subtotal nephrectomized ( STNx ) rats were randomly assigned to receive vehicle ( n=10 ) or the P12821 inhibitor perindopril ( 8 mg/l drinking water ) for 12 weeks duration ( n=10 ) . Sham-operated rats were used as controls ( n=10 ) . Glomerular capillary endothelial cell density was evaluated by immunostaining for the pan-endothelial cell marker Q06609 -1 and P15692 expression was assessed by quantitative in situ hybridization . RESULTS : In STNx rats glomerular capillary endothelial cell density was reduced to 19 % that of sham rats ( P < 0.01 ) with a concomitant reduction in glomerular P15692 expression , also to 19 % of sham rats ( P < 0.01 ) . DB00790 treatment was associated with normalization of both capillary endothelial cell density and glomerular P15692 mRNA . CONCLUSIONS : Reduction in glomerular P15692 expression is a feature of the renal pathology that follows subtotal nephrectomy . In the context of the known functions of this growth factor , these findings suggest that diminution in P15692 may contribute to the demonstrated loss of glomerular endothelium that develops in this model of progressive renal disease . P00797 -angiotensin system modulation : the weight of evidence . Modulation of the renin-angiotensin system is considered to be the most complete way to manage high-risk patients including those with hypertension . P12821 ( P12821 ) inhibitors are effective at reducing the morbidity and mortality of patients with overt clinical heart failure , asymptomatic left ventricular dysfunction , and uncomplicated myocardial infarction . Furthermore , recent trials like the Heart Outcomes Prevention Evaluations ( HOPE ) study and the EUropean trial on Reduction Of cardiac events with DB00790 in stable coronary Artery disease ( EUROPA ) support extending the use of P12821 inhibitors to the routine/first-line treatment of patients with an increased global cardiovascular risk . Although some investigators have seen the development of angiotensin II receptor blockers ( ARBs ) as a more effective and tolerable way of reproducing the benefits of P12821 inhibition , there remain important concerns regarding the distinct pharmacologic profiles and modes of action of these two classes of drugs . Careful evaluation of data from recent large-scale studies revealed that , unlike P12821 inhibitors , ARBs are either neutral or may actually increase rates of myocardial infarction despite similar levels of blood pressure reduction . The fact that this effect is most apparent when ARBs are compared with placebo in the absence of concomitant P12821 inhibitors suggests that differential effects on the angiotensin II type 2 ( AT(2) ) receptors may be important . Other important pharmacologic differences are also known to be present and may be of direct relevance . The weight of available evidence therefore supports the use of appropriate P12821 inhibitor regimens , although not ARBs , in the treatment of global cardiovascular risk . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Early life stress changes concentrations of neuropeptide Y and corticotropin-releasing hormone in adult rat brain . DB01356 treatment modifies these changes . Experiences of early life stress are more prevalent among depressed patients than healthy controls . Neuropeptide Y ( P01303 ) was suggested to play a role in the pathophysiology of depression . Consequently , we investigated in adult rats the effects of maternal deprivation for 3 h/day during postnatal days ( P01160 ) 2-14 and of dietary lithium during P01160 50-83 on brain levels of P01303 -like immunoreactivity ( LI ) . Brain levels of corticotropin-releasing hormone ( P06850 ) and serum corticosterone were also measured . Maternal deprivation reduced P01303 -LI levels in the hippocampus and the striatum but increased P01303 -LI and P06850 -LI levels in the hypothalamus . DB01356 treatment counteracted the effect of maternal deprivation in the hippocampus and striatum by increasing P01303 -LI levels . In the hypothalamus , lithium tended to decrease P06850 -LI but further increased levels of P01303 -LI ; it also increased serum corticosterone levels . The results suggest that early life stress has long-term effects on brain P01303 with implications for the development of depression/vulnerability to stress , and that one therapeutic mechanism of action of lithium is to increase brain P01303 . DB06643 -- an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 ) , a cytokine member of the P01375 family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis . Novel replication-incompetent adenoviral B-group vectors : high vector stability and yield in O15534 . P13671 cells . Adenoviral vectors based on adenovirus type 35 ( rAd35 ) have the advantage of low natural vector immunity and induce strong , insert-specific T- and B-cell responses , making them prime-candidate vaccine carriers . However , severe vector-genome instability of E1-deleted rAd35 vectors was observed , hampering universal use . The instability of E1-deleted rAd35 vector proved to be caused by low pIX expression induced by removal of the pIX promoter , which was located in the E1B region of B-group viruses . Reinsertion of a minimal pIX promoter resulted in stable vectors able to harbour large DNA inserts ( > 5 kb ) . In addition , it is shown that replacement of the E4-Orf6 region of Ad35 by the E4-Orf6 region of Ad5 resulted in successful propagation of an E1-deleted rAd35 vector on existing E1-complementing cell lines , such as O15534 . P13671 cells . The ability to produce these carriers on O15534 . P13671 contributes significantly to the scale of manufacturing of rAd35-based vaccines . Next , a stable rAd35 vaccine was generated carrying Mycobacterium tuberculosis antigens Ag85A , Ag85B and TB10.4 . The antigens were fused directly , resulting in expression of a single polyprotein . This vaccine induced dose-dependent P01730 + and CD8+ T-cell responses against multiple antigens in mice . It is concluded that the described improvements to the rAd35 vector contribute significantly to the further development of rAd35 carriers for mass-vaccination programmes for diseases such as tuberculosis , AIDS and malaria . Diurnal alterations in circadian genes and peptides in major depressive disorder before and after escitalopram treatment . BACKGROUND : Strong links exist between circadian disturbances and some of the most characteristic symptoms of clinical major depressive disorder ( MDD ) . However , changes in the expression of clock genes or neuropeptides related to the regulation of circadian rhythm that may influence the susceptibility to recurrence after antidepressant treatment in MDD have not been investigated . METHODS : Blood samples were collected at 4h intervals for 24h from 12 male healthy controls and 12 male MDD patients before and after treatment with escitalopram for 8 weeks . The outcome measures included the relative expression of clock gene mRNA ( PERIOD1 , PERIOD2 , PERIOD3 , Q16526 , O00327 , Q99743 , and GSK-3β ) , and the levels of serum melatonin , vasoactive intestinal polypeptide ( P01282 ) , cortisol , adrenocorticotropic hormone ( DB01285 ) , insulin-like growth factor-1 ( DB01277 ) , and growth hormone ( GH ) . RESULTS : Compared with healthy controls , MDD patients showed disruptions in the diurnal rhythms of the expression of PERIOD1 , PERIOD2 , Q16526 , O00327 , Q99743 , and GSK-3β and disruptions in the diurnal rhythms of the release of melatonin , P01282 , cortisol , DB01285 , DB01277 , and GH . Several of these disruptions ( i.e. , O15534 , Q16526 , melatonin , P01282 , cortisol , DB01285 , and DB01277 ) persisted 8 weeks after escitalopram treatment , similar to the increase in the 24h levels of P01282 and decreases in the 24h levels of cortisol and DB01285 . CONCLUSION : These persistent neurobiological changes may play a role in MDD symptoms that are thought to contribute to the vulnerability to recurrence and long-term maintenance therapy . [ Changes in chemokine receptor 4 , interleukin-6 , and collagen X expression in the ATDC5 cell line stimulated by cyclic tensile strain and stromal cell-derived factor-1 ] . OBJECTIVE : This study further explores the stromal cell-derived factor-1 ( P48061 ) /chemokine receptor 4 ( P61073 ) signaling axis mechanism in temporomandibular joint osteoarthritis ( OA ) by detecting the changes in P61073 , interleukin ( IL ) -6 , and collagen X expression in the ATDC5 cell line stimulated by the cyclic tensile strain and P48061 . METHODS : P01308 -transferrin-selenium ( ITS ) was used to induce ATDC5 cells to differentiate into chondrocyte-like cells . After three weeks , the cells were divided into two groups : those with and without cyclic tensile strain . These groups were further divided into the negative control and P48061 groups . Strain force of 20 % was applied . After 12 h , the total proteins were extracted from cells of the four groups , and Western blot analysis was used to detect the changes in P61073 , P05231 , and collagen X expression . RESULTS : P48061 could enhance P61073 , P05231 , and collagen X expressions in the chondrocytes , and 20 % tensile strain force could further upregulate the three factors . CONCLUSION : Under abnormal tensile force , P48061 can upregulate its specific receptor P61073 , thus increasing its-binding efficiency and resulting in the activation of the P48061 / P61073 axis . This condition enhances the expressions of P05231 and other inflammatory factors and directly damages to cartilage tissue . Such damage directly promotes chondrocyte hypertrophy , which enhances collagen X expression . DB00790 : possible use in cancer therapy . Since angiogenesis is essential for the growth of any solid tumor , emerging efforts are being made to develop antiangiogenic therapy . To date , however , no antiangiogenic agent has become widely available for the clinical setting . Angiotensin I-converting enzyme ( P12821 ) inhibitors are commonly used as antihypertensive agents and it has recently been suggested that they decrease the risk of cancer . Studies have found that an P12821 inhibitor , perindopril , is a potent inhibitor of experimental tumor development and angiogenesis at a clinically comparable dose . The potent angiogenic factor , vascular endothelial growth factor ( P15692 ) , is significantly suppressed by perindopril and also inhibits P15692 -induced tumor growth . In vitro studies showed that perindopril is not cytotoxic to either tumor cells or endothelial cells . Since perindopril is already in widespread clinical use without serious side effects , it may represent a potential new strategy for anticancer therapy . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Investigation of immunomodulatory properties of human Wharton 's Jelly-derived DB05914 after lentiviral transduction . Human Wharton 's Jelly-derived Mesenchymal Stem Cells ( hWJ-MSCs ) are considered as an alternative for bone-marrow-derived MSCs . These cells have immunosuppressive properties . It was unclear whether the WJ-MSCs would sustain their immunomodulatory characteristics after lentiviral transduction or not . In this study , we evaluated immunomodulatory properties of WJ-MSCs after lentiviral transduction . HWJ-MSCs were transduced with lentiviral particles . Expression of transduced and un-transduced hWJ-MSCs surface molecules and secretion of P22301 , P14210 , P15692 and TGF-β was analyzed . Cell proliferation and frequency of P01730 (+)CD25(+) CD127(low/neg) Foxp3(+) T regulatory cells was measured . There was no difference between the surface markers and secretion of P22301 , P14210 , P15692 and TGF-β in transduced and un-transduced hWJ-MSCs . Both cells inhibited the proliferation of PHA stimulated PBMCs , and improved the frequency of T regulatory cells . These findings suggest that lentiviral transduction does not alter the immunomodulatory function of hWJ-MSCs . However , lentiviral transduction may have a wide range of applications in gene therapy . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . Vascular endothelial growth factor receptors in osteoclast differentiation and function . Osteoclasts are derived from haematopoietic stem cell precursors of the monocyte/macrophage cell lineage , through interaction with factors that are believed to include P09603 and O14788 . P15692 is a proangiogenic cytokine that has been shown to promote osteoclast differentiation and survival . In this study , we assessed the role of P15692 and its receptors in osteoclastogenesis , in vitro , by culturing osteoclast precursors in the presence of P15692 , P15692 receptor-specific ligands , and blocking antibodies to P15692 receptors . Activation of P17948 in the presence of O14788 induces osteoclast differentiation . Stimulating the receptors individually induced increased resorption by osteoclasts compared to controls but not to the level observed when stimulating both receptors simultaneously . We have shown that P15692 induces osteoclast differentiation through its action on P17948 . The way in which P15692 mediates its effect on mature osteoclast activity , however , may be through its interaction with both receptor subtypes . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Genetic basis of delay discounting in frequent gamblers : examination of a priori candidates and exploration of a panel of dopamine-related loci . INTRODUCTION : Delay discounting is a behavioral economic index of impulsivity that reflects preferences for small immediate rewards relative to larger delayed rewards . It has been consistently linked to pathological gambling and other forms of addictive behavior , and has been proposed to be a behavioral characteristic that may link genetic variation and risk of developing addictive disorders ( i.e. , an endophenotype ) . Studies to date have revealed significant associations with polymorphisms associated with dopamine neurotransmission . The current study examined associations between delay discounting and both previously linked variants and a novel panel of dopamine-related variants in a sample of frequent gamblers . METHODS : Participants were 175 weekly gamblers of European ancestry who completed the Monetary Choice Questionnaire to assess delay discounting preferences and provided a DNA via saliva . RESULTS : In a priori tests , two loci previously associated with delayed reward discounting ( rs1800497 and rs4680 ) were not replicated , however , the long form of P21917 VNTR was significantly associated with lower discounting of delayed rewards . Exploratory analysis of the dopamine-related panel revealed 11 additional significant associations in genes associated with dopamine synthesis , breakdown , reuptake , and receptor function ( P35462 , Q01959 , DDC , P09172 , and Q05940 ) . An aggregate genetic risk score from the nominally significant loci accounted for 17 % of the variance in discounting . Mediational analyses largely supported the presence of indirect effects between the associated loci , delay discounting , and pathological gambling severity . CONCLUSIONS : These findings do not replicate previously reported associations but identify several novel candidates and provide preliminary support for a systems biology approach to understand the genetic basis of delay discounting . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . Role of angiotensin II in the remodeling induced by a chronic increase in flow in rat mesenteric resistance arteries . Angiotensin II is a potent growth factor involved in arterial wall homeostasis . In resistance arteries , chronic increases in blood flow induce a rise in diameter associated with arterial wall hypertrophy . Nevertheless , the role of angiotensin II in this remodeling is unknown . We investigated the effect of blocking angiotensin II production or receptor activation on flow-induced remodeling of mesenteric resistance arteries . Arteries were ligated in vivo to generate high-flow arteries compared with normal flow ( control ) vessels located at a distance . Arteries were isolated after 1 week for in vitro analysis . Arterial diameter , media surface , endothelial NO synthase expression , superoxide production , and extracellular signal-regulated kinase 1/2 phosphorylation were higher in high-flow than in control arteries . P12821 inhibition ( perindopril ) and angiotensin II type 1 receptor blockade ( candesartan ) prevented arterial wall hypertrophy without affecting diameter enlargement . The nonselective vasodilator hydralazine had no effect on remodeling . Although perindopril and candesartan increased endothelial NO synthase expression in high-flow arteries , hypertrophy remained in rats treated with N(G)-nitro-l-arginine methyl ester and mice lacking endothelial NO synthase . DB00790 and candesartan reduced oxidative stress in high-flow arteries , but superoxide scavenging did not prevent hypertrophy . Both Tempol and the absence of endothelial NO synthase prevented the rise in diameter in high-flow vessels . P27361 /2 activation in high-flow arteries was prevented by perindopril and candesartan and not by hydralazine . P27361 /2 inhibition in vivo ( U0126 ) prevented hypertrophy in high-flow arteries . Thus , a chronic rise in blood flow in resistance arteries induces a diameter enlargement involving NO and superoxide , whereas hypertrophy was associated with extracellular signal-regulated kinase 1/2 activation by angiotensin II . DB00790 decreases P wave dispersion in patients with stage 1 hypertension . INTRODUCTION : P12821 inhibitors prevent atrial fibrillation episodes by effective control of blood pressure and improving electrical and structural remodelling in the atria . Increased P wave dispersion ( P35670 ) is a non-invasive electrocardiographic marker for paroxysmal atrial fibrillation . The aim of the study was to evaluate the effect of perindopril treatment on P35670 in hypertensive patients . METHODS : Forty-eight hypertensive patients ( mean age 57.4+/-11.8 years , 18 men ) were included . Blood pressure values were determined and 12-lead electrocardiograms were recorded at the beginning and at the first week , first month , third month and sixth month of the perindopril treatment.The difference between maximum and minimum P wave durations was calculated as P35670 . RESULTS : PWDs were significantly shortened at the first , third and sixth months ( 41.7+/-8.8 ms , Q04695 +/-6.9 ms and 38.3+/-7.1 ms , respectively ) compared with baseline and first-week measurements ( 54.3+/-9.2 ms and 49.0+/-9.1 ms , respectively , p < 0.001 ) . Baseline P35670 was correlated with body mass index ( r=0.32 , p=0.026 ) , while P35670 at the sixth month of treatment was significantly correlated with left atrial volume index ( r=0.30 , p=0.042 ) . Multiple linear regression analysis revealed that P35670 at the sixth month was related to baseline P35670 ( p=0.001 ) . CONCLUSION : DB00790 treatment significantly reduced P35670 in hypertensive patients . Inhibition of central angiotensin converting enzyme ameliorates scopolamine induced memory impairment in mice : role of cholinergic neurotransmission , cerebral blood flow and brain energy metabolism . Evidences indicate that inhibition of central P00797 angiotensin system ( DB01367 ) ameliorates memory impairment in animals and humans . Earlier we have reported involvement of central angiotensin converting enzyme ( P12821 ) in streptozotocin induced neurodegeneration and memory impairment . The present study investigated the role of central P12821 in cholinergic neurotransmission , brain energy metabolism and cerebral blood flow ( Q03701 ) in model of memory impairment induced by injection of scopolamine in mice . DB00790 ( 0.05 and 0.1 mg/kg , PO ) was given orally for one week before administration of scopolamine ( 3mg/kg , IP ) . Then , memory function was evaluated by Morris water maze and passive avoidance tests . Q03701 was measured by laser Doppler flowmetry . Biochemical and molecular parameters were estimated after the completion of behavioral studies . DB00747 caused impairment in memory which was associated with reduced Q03701 , acetylcholine ( ACh ) level and elevated acetylcholinesterase ( P22303 ) activity and malondialdehyde ( MDA ) level . DB00790 ameliorated scopolamine induced amnesia in both the behavioral paradigms . Further , perindopril prevented elevation of P22303 and MDA level in mice brain . There was a significant increase in Q03701 and ACh level in perindopril treated mice . However , scopolamine had no significant effect on DB00171 level and mRNA expression of angiotensin receptors and P12821 in cortex and hippocampus . But , perindopril significantly decreased P12821 activity in brain without affecting its mRNA expression . The study clearly showed the interaction between P12821 and cholinergic neurotransmission and beneficial effect of perindopril can be attributed to improvement in central cholinergic neurotransmission and Q03701 . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . β-cryptoxanthin regulates bone resorption related-cytokine production in human periodontal ligament cells . OBJECTIVE : β-cryptoxanthin ( β-cry ) is a type of carotenoid found in certain fruits and vegetables . Although it has been shown that β-cry inhibits alveolar bone resorption , the molecular mechanisms for this have not yet been clarified . In the present study , we investigated the effects of β-cry on bone resorption related-cytokine production in human periodontal ligament ( hPDL ) cells . DESIGN : hPDL cells were stimulated with β-cry ( 1×10(-7)mol/l ) , mechanical stress ( 1 or 6MPa ) , and P. gingivalis . The production of interleukin ( IL ) -1β , P05231 , P10145 , tumour necrosis factor ( P01375 ) -α , osteoprotegerin ( O00300 ) , and receptor activator of nuclear factor kappa-B ligand ( O14788 ) were analyzed by RT-PCR and ELISA . RESULTS : The production of IL-1β , P05231 , P10145 , and P01375 -α was not induced in hPDL cells after stimulation with β-cry , although these cytokines were produced after stimulation with P. gingivalis . On the other hand , P05231 and P10145 were produced after exposure to 6MPa of mechanical stress . The production of P05231 and P10145 was significantly decreased by the addition of β-cry . Furthermore , β-cry up-regulated the production of O00300 , but not O14788 . CONCLUSION : β-cry inhibited the production of P05231 and P10145 induced by mechanical stress and periodontopathogenic bacteria in hPDL cells . Moreover , β-cry up-regulated O00300 production . These results suggest that β-cry may prevent bone resorption in periodontitis . Synthesis and application in polypropylene of a novel of phosphorus-containing intumescent flame retardant . A novel phosphorus-containing triazine oligomer poly ( 2-morpholinyl-4-penta-erythritol phosphate-1,3,5-triazine ) ( PMPT ) was synthesized as a kind of tri-component intumescent flame retardant ( IFR ) . The chemical structure of PMPT was characterized by FTIR , 1H-NMR and 31P-NMR , and the mechanical and flammability properties of FR-PP were measured . The FTIR results showed that the expected chemical reactions had happened at each step . The 1H-NMR and 31P-NMR spectra also agreed with the chemical structure of PMPT . The slight effect of PMPT on the mechanical properties of FR-PP suggested that PMPT and PP are compatible . The high limited oxygen index ( LOI ) values of FR-PP revealed that PMPT was an efficient IFR and there was the synergistic effect between PMPT and ammonium polyphosphate/ pentaerythritol ( P05067 / O15534 ) . Eupolyphaga sinensis walker displays inhibition on hepatocellular carcinoma through regulating cell growth and metastasis signaling . Tumor growth and metastasis are responsible for most cancer patients ' deaths . Here , we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis . Compared with proliferation , colony formation , transwell assay and transplantable tumor in nude mouse in vitro and vivo , eupolyphaga sinensis walker extract ( ESWE ) showed good inhibition on the SMMC-7721 cell growth and metastasis . Using genome-wide microarray analysis , we found the down-regulated growth and metastasis factors , and selected down-regulated genes were confirmed by real-time PCR . Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE . Moreover , our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ , Akt , m-TOR , Erk1/2 , MEK-2 , Raf and JNK-1 , but also increasing cyclin D1 protein levels and decreasing amount of cyclin E , cyclin B1 and cdc2 of the cycle proteins . At the same time , ESWE reduced P08253 , P14780 and P61073 , P00747 , NFκB and P04637 activities . Overall , our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC , AKT , MAPK signaling and related metastasis signaling , having potential in cancer therapy . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . Impaired urine concentration and absence of tissue P12821 : involvement of medullary transport proteins . P12821 .2 mice lack all tissue angiotensin-converting enzyme ( P12821 ) but have 33 % of normal plasma P12821 activity . They exhibit the urine-concentrating defect and hyperkalemia present in mice that lack all P12821 , but in contrast to the complete knockout , P12821 .2 mice have normal medullary histology and creatinine clearance . To explore the urine-concentrating defect in P12821 .2 mice , renal medullary transport proteins were analyzed using Western blot analysis . In the inner medulla , UT-A1 , P51800 , and aquaporin-1 ( P29972 ) were significantly reduced to 28 +/- 5 , 6 +/- 6 , and 39 +/- 5 % of the level in wild-type mice , respectively , whereas P41181 and UT-B were unchanged . In the outer medulla , Na(+)-K(+)-2Cl(-) cotransporter ( Q13621 /BSC1 ) and P29972 were significantly reduced to 56 +/- 11 and 29 +/- 6 % , respectively , whereas Na(+)-K(+)-ATPase , UT-A2 , UT-B , and P41181 were unchanged , and renal outer medullary potassium channel was significantly increased to 711 +/- 187 % of the level in wild-type mice . The abnormal expression of these transporters was similar in P12821 .2 mice backcrossed onto a C57BL/6 or a Swiss background and was not rescued by P03950 II infusion . We conclude that the urine-concentrating defect in P12821 .2 mice is associated with , and may result from , downregulation of some or all of these key urea , salt , and water transport proteins . A role for plasma transforming growth factor-beta and matrix metalloproteinases in aortic aneurysm surveillance in Marfan syndrome ? BACKGROUND : We have previously shown that the angiotensin-converting enzyme ( P12821 ) inhibitor perindopril reduced aortic diameter by 3-7mm in Marfan syndrome ( MFS ) patients . Excessive signalling by the transforming growth factor-beta ( TGF-beta ) has been implicated in the development of aortic dilatation . We hypothesised that reduction in aortic diameter would correlate with reduction in plasma TGF-beta and matrix metalloproteinase ( MMP ) levels . METHODS : 17 MFS patients ( aged 33+/-5 ( mean+/-SD ) ) on standard beta-blocker therapy were randomised to also receive perindopril ( n=10 ) or placebo ( n=7 ) for 24 weeks in a double blind study . Aortic root diameters were assessed at four sites via transthoracic echocardiography . Venous blood samples were analysed for latent and active TGF-beta , P08253 and P08254 levels . RESULTS : DB00790 significantly reduced aortic root diameters relative to placebo in both end-systole and end-diastole ( by 1.2-3mm/m(2) , p < 0.001 ) . In addition , compared to placebo perindopril significantly reduced latent TGF-beta levels by 14.0+/-4.5ng/ml ( p=0.01 ) , active TGF-beta levels by 4+/-1ng/ml ( p=0.02 ) , P08253 levels by 22+/-6ng/ml ( p < 0.001 ) , and P08254 levels by 5+/-1ng/ml ( p < 0.001 ) . There were moderately strong correlations between the pre/post intervention change in aortic diameters and the change in both latent ( r=0.49-0.76 , p=0.001-0.04 ) and active TGF-beta ( r=0.59-0.73 , p=0.002-0.02 ) , P08253 ( r=0.63-0.75 , p=0.001-0.007 ) , and P08254 plasma levels ( r=0.81-0.83 , p < 0.0001 ) . CONCLUSIONS : Plasma TGF-beta , P08253 and P08254 should be further explored in longitudinal trials as potential prognostic indicators of progression of aortic dilatation and response to therapy in MFS . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy . Sustained cardiac pressure overload induces hypertrophy and pathological remodeling , frequently leading to heart failure . Genetically engineered hyperstimulation of guanosine 3',5'-cyclic monophosphate ( cGMP ) synthesis counters this response . Here , we show that blocking the intrinsic catabolism of cGMP with an oral phosphodiesterase-5A ( O76074 ) inhibitor ( sildenafil ) suppresses chamber and myocyte hypertrophy , and improves in vivo heart function in mice exposed to chronic pressure overload induced by transverse aortic constriction . DB00203 also reverses pre-established hypertrophy induced by pressure load while restoring chamber function to normal . cGMP catabolism by O76074 increases in pressure-loaded hearts , leading to activation of cGMP-dependent protein kinase with inhibition of O76074 . O76074 inhibition deactivates multiple hypertrophy signaling pathways triggered by pressure load ( the calcineurin/NFAT , phosphoinositide-3 kinase ( PI3K ) /Akt , and P27361 /2 signaling pathways ) . But it does not suppress hypertrophy induced by overexpression of calcineurin in vitro or Akt in vivo , suggesting upstream targeting of these pathways . O76074 inhibition may provide a new treatment strategy for cardiac hypertrophy and remodeling . The dopamine-4 receptor gene associated with binge eating and weight gain in women with seasonal affective disorder : an evolutionary perspective . BACKGROUND : We recently described a preliminary association between the hypofunctional seven-repeat allele of the dopamine-4 receptor gene ( P21917 ) and increased maximal lifetime body mass index in women with seasonal affective disorder ( SAD ) . In this study , we examined whether binge eating behavior mediated this putative association . METHODS : The study sample consisted of 131 women with winter SAD who reported increased intake of high-carbohydrate/high-fat foods during depressive episodes . We compared rates of binge eating behavior in the two genotypic groups defined by the presence or absence of the seven-repeat allele of P21917 . RESULTS : Consistent with our working hypothesis , the proportion of binge eaters was significantly greater in probands with the seven-repeat allele ( 18 of 46 , Q04695 % ) than in probands without this allele ( 14 of 85 , 16.5 % ) [ chi(2)(1)= 8.32 , p = .004 ; odds ratio = 3.25 , 95 % confidence interval 1.43 , 7.41 ] . CONCLUSIONS : Pending replication in other samples , these results point to a genetic factor that could help in the early identification and treatment of women at higher risk for seasonal weight gain associated with binge eating behavior . At a theoretic level , the current results suggest a novel link between evolutionary models of seasonal weight gain on the one hand and the P21917 gene on the other .

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interacts_with DB00368?

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Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . [ Polymorphisms of 2B-adrenergic receptor and endothelial NO-Synthase genes in genesis of the hereditary sick sinus node syndrome ] . In this work we have demonstrated for the first time on the clinico-genetic material association between hereditary sick sinus node syndrome ( SSNS ) P18089 and P29474 genes polymorphisms . We have established predominance of homozygote genotype of more rare DD allele in patients with SSNS ( 28 % ) compared with subjects of control group ( 8.99 % ) . We have found predominance of heterozygote genotype 4a/4b in patients with SSNS compared with subjects of control group ( 41.8 and 25.39 % , respectively ) . The data obtained allow to suggest that P29474 gene polymorphism might be associated with SSNS . Role of tyrosyl-DNA phosphodiesterase 1 and inter-players in regulation of tumor cell sensitivity to topoisomerase I inhibition . Q9NUW8 ( Q9NUW8 ) plays a unique function as it catalyzes the repair of topoisomerase I-mediated DNA damage . Thus , ovarian carcinoma cell lines exhibiting increased Q9NUW8 levels and resistance to the topoisomerase I poisons campthotecins were used to clarify the role of this enzyme . The camptothecin gimatecan was employed as a tool to inhibit topoisomerase I because it produces a persistent damage . The resistant sublines displayed an increased capability to repair drug-induced single-strand breaks and a reduced amount of drug-induced double-strand breaks , which was enhanced following Q9NUW8 silencing . In loss of function studies using U2-OS cells , we found that Q9NUW8 knockdown did not produce a change in sensitivity to camptothecin , whereas co-silencing of other pathways cooperating with Q9NUW8 in cell response to topoisomerase I poisons indicated that P18887 and P38398 were major regulators of sensitivity . No change in cellular sensitivity was observed when Q9NUW8 was silenced concomitantly to O75943 , which participates in the stabilization of collapsed replication forks . The expression of dominant-negative P09874 in cells with reduced expression of Q9NUW8 due to a constitutively expressed Q9NUW8 targeting microRNA did not modulate cell sensitivity to camptothecin . Mild resistance to gimatecan was observed in cells over-expressing Q9NUW8 , a feature associated with decreased levels of drug-induced single-strand breaks . In conclusion , since Q9NUW8 alone can account for mild levels of camptothecin resistance , repair of topoisomerase I-mediated DNA damage likely occurs through redundant pathways mainly implicating P38398 and P18887 , but not O75943 and P09874 . These findings may be relevant to define novel therapeutic strategies . Acute erythropoietin cardioprotection is mediated by endothelial response . Increasing evidence indicates that high levels of serum erythropoietin ( Epo ) can lessen ischemia-reperfusion injury in the heart and multiple cardiac cell types have been suggested to play a role in this Epo effect . To clarify the mechanisms underlying this cardioprotection , we explored Epo treatment of coronary artery endothelial cells and Epo cardioprotection in a Mus musculus model with Epo receptor expression restricted to hematopoietic and endothelial cells ( ΔEpoR ) . Epo stimulation of coronary artery endothelial cells upregulated endothelial nitric oxide synthase ( P29474 ) activity in vitro and in vivo , and enhanced nitric oxide ( NO ) production that was determined directly by real-time measurements of gaseous NO release . Epo stimulated phosphoinositide 3-kinase ( PI3K ) /protein kinase B ( AKT ) and mitogen-activated protein kinase kinase ( MEK ) /extracellular signal regulated kinase ( P29323 ) signaling pathways , and inhibition of PI3K , but not MEK activity , blocked Epo-induced NO production . To verify the potential of this Epo effect in cardioprotection in vivo , ΔEpoR-mice with Epo response in heart restricted to endothelium were treated with Epo . These mice exhibited a similar increase in P29474 phosphorylation in coronary artery endothelium as that found in wild type ( WT ) mice . In addition , in both WT- and ΔEpoR-mice , exogenous Epo treatment prior to myocardial ischemia provided comparable protection . These data provide the first evidence that endothelial cell response to Epo is sufficient to achieve an acute cardioprotective effect . The immediate response of coronary artery endothelial cells to Epo stimulation by NO production may be a critical mechanism underlying this Epo cardioprotection . P18089 gene insertion/deletion polymorphism and artery compliance . BACKGROUND : The P18089 gene insertion/deletion ( I/D ) polymorphism is associated with various cardiovascular and metabolic phenotypes . Large ( C1 ) and small ( P06681 ) artery compliance , assessed by pulse wave analysis , is considered as sensitive markers or risk factors for cardiovascular disease . Therefore whether the P18089 I/D polymorphism is associated with C1 and P06681 need to be investigated . METHODS : A total of 227 men and 243 women were enrolled in a Chinese family-based study . C1 and P06681 were measured by pulse wave analysis . P18089 genotypes were determined by polymerase chain reaction . Statistical methods included generalized estimation equations and quantitative transmission disequilibrium test . RESULTS : The II ( 31.9 % ) , ID ( 46.8 % ) and DD ( 21.3 % ) genotype frequencies were in Hardy-Weinberg equilibrium ( P = 0.73 ) . The covariates selected by stepwise regression for C1 and P06681 were age , systolic pressure and gender . The population based association analysis showed that C1 and P06681 were not associated with P18089 genotype both before ( C1 : P = 0.28 ; P06681 : P = 0.27 ) and after ( C1 : P = 0.58 ; P06681 : P = 0.18 ) the adjustment . The family-based analyses of 128 informative offspring showed that transmission of the D-allele was not associated with C1 or P06681 , both before ( C1 : P = 0.42 ; P06681 : P = 0.85 ) and after ( C1 : P = 0.31 ; P06681 : P = 0.82 ) the adjustment . CONCLUSION : The study do not support that the P18089 gene I/D polymorphism has a major gene effect on C1 or P06681 in the Chinese population of current sample size . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Candidate gene polymorphisms in the serotonergic pathway : influence on depression symptomatology in an elderly population . BACKGROUND : Depressed mood is a major concern in the elderly , with consequences for morbidity and mortality . Previous studies have demonstrated that genetic factors in depression and subsyndromal depressive symptoms are no less important in the elderly than during other life stages . Variations in genes included in the serotonin system have been suggested as risk factors for various psychiatric disorders but may also serve as candidates for normal variations in mood . METHODS : This study included 684 elderly Danish twins to investigate the influence of 11 polymorphisms in 7 serotonin system genes on the mean level of depression symptomatology assessed over several years , reflecting individuals ' underlying mood level . RESULTS : A suggestive association of sequence variations in genes responsible for the synthesis ( P17752 ) , recognition ( 5- P28223 ) , and degradation ( P21397 ) of serotonin with depression symptomatology was found , although the effect was generally restricted to men . We also found that a specific haplotype in Q05940 , the gene encoding the vesicular monoamine transporter , was significantly associated with depression symptoms in men ( p= .007 ) . CONCLUSIONS : These results suggest that variations in genes encoding the components of serotonin metabolism may influence the basic mood level and that different genetic factors may apply in men and women . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Modulation of a number of genes on personality traits in a sample of healthy subjects . A large number of studies investigated the genetic modulation of personality with mixed results . As a confirmatory analysis of previous findings , we firstly examined the association between several previously examined single nucleotide polymorphisms ( SNPs ) and personality traits in a sample of 158 healthy subjects . As a secondary aim , we tested the potential modulation of additional never previously investigated genes on personality . A blood sample was collected and the Temperament and Character Inventory ( TCI ) has been administered to all participants . Multivariate analysis of covariance , controlling for sex and age , was used to test SNP influence on TCI scores . Examination of previously studied gene variants showed an effect of adrenergic alpha 2B receptor ( P18089 ) on Cooperativeness and of serotonin receptor P28223 on Self Directedness . Examination of new variants revealed that sex hormone binding protein ( P04278 ) was associated with reward dependence . Moreover , several additional variants showed a tendency towards association with some TCI traits , confirming previous results . This study suggests that P18089 , P28223 and P04278 genes may be involved in the modulation of personality in healthy subjects . The major limitation of this study was the small sample size . Alpha 2B adrenoceptor genotype moderates effect of reboxetine on negative emotional memory bias in healthy volunteers . Evidence suggests that emotional memory plays a role in the pathophysiology of depression/anxiety disorders . DB00368 crucially modulates emotional memory . Genetic variants involved in noradrenergic signaling contribute to individual differences in emotional memory and vulnerability to psychopathology . A functional deletion polymorphism in the α-2B adrenoceptor gene ( P18089 ) has been linked to emotional memory and post-traumatic stress disorder . The noradrenaline reuptake inhibitor reboxetine attenuates enhanced memory for negative stimuli in healthy and depressed individuals . We examined whether the effect of reboxetine on emotional memory in healthy individuals would be moderated by P18089 genotype . P18089 deletion carriers demonstrated enhanced emotional memory for negative stimuli compared with deletion noncarriers , consistent with prior studies . DB00234 attenuated enhanced memory for negative stimuli in deletion noncarriers but had no significant effect in deletion carriers . This is the first demonstration of genetic variation influencing antidepressant drug effects on emotional processing in healthy humans . Molecular systematics of armadillos ( Xenarthra , Dasypodidae ) : contribution of maximum likelihood and Bayesian analyses of mitochondrial and nuclear genes . The 30 living species of armadillos , anteaters , and sloths ( Mammalia : Xenarthra ) represent one of the three major clades of placentals . Armadillos ( Cingulata : Dasypodidae ) are the earliest and most speciose xenarthran lineage with 21 described species . The question of their tricky phylogeny was here studied by adding two mitochondrial genes ( P03886 [ P03886 ] and 12S ribosomal RNA [ 12S rRNA ] ) to the three protein-coding nuclear genes ( alpha2B adrenergic receptor [ P18089 ] , breast cancer susceptibility exon 11 [ P38398 ] , and P04275 exon 28 [ P04275 ] ) yielding a total of 6869 aligned nucleotide sites for thirteen xenarthran species . The two mitochondrial genes were characterized by marked excesses of transitions over transversions-with a strong bias toward CT transitions for the 12S rRNA-and exhibited two- to fivefold faster evolutionary rates than the fastest nuclear gene ( P18089 ) . Maximum likelihood and Bayesian phylogenetic analyses supported the monophyly of Dasypodinae , Tolypeutinae , and Euphractinae , with the latter two armadillo subfamilies strongly clustering together . Conflicting branching points between individual genes involved relationships within the subfamilies Tolypeutinae and Euphractinae . Owing to a greater number of informative sites , the overall concatenation favored the mitochondrial topology with the classical grouping of Cabassous and Priodontes within Tolypeutinae , and a close relationship between Euphractus and Chaetophractus within Euphractinae . However , low statistical support values associated with almost equal distributions of apomorphies among alternatives suggested that two parallel events of rapid speciation occurred within these two armadillo subfamilies . P18089 genotype differentially modulates stress-induced neural activity in the amygdala and hippocampus during emotional memory retrieval . RATIONALE : DB00368 interacts with stress hormones in the amygdala and hippocampus to enhance emotional memory consolidation , but the noradrenergic-glucocorticoid interaction at retrieval , where stress impairs memory , is less understood . OBJECTIVES : We used a genetic neuroimaging approach to investigate whether a genetic variation of the noradrenergic system impacts stress-induced neural activity in amygdala and hippocampus during recognition of emotional memory . METHODS : This study is based on genotype-dependent reanalysis of data from our previous publication ( Li et al. Brain Imaging Behav 2014 ) . Twenty-two healthy male volunteers were genotyped for the P18089 gene encoding the α2B-adrenergic receptor . Ten deletion carriers and 12 noncarriers performed an emotional face recognition task , while their brain activity was measured with fMRI . During encoding , 50 fearful and 50 neutral faces were presented . One hour later , they underwent either an acute stress ( Trier Social Stress Test ) or a control procedure which was followed immediately by the retrieval session , where participants had to discriminate between 100 old and 50 new faces . RESULTS : A genotype-dependent modulation of neural activity at retrieval was found in the bilateral amygdala and right hippocampus . Deletion carriers showed decreased neural activity in the amygdala when recognizing emotional faces in control condition and increased amygdala activity under stress . Noncarriers showed no differences in emotional modulated amygdala activation under stress or control . Instead , stress-induced increases during recognition of emotional faces were present in the right hippocampus . CONCLUSION : The genotype-dependent effects of acute stress on neural activity in amygdala and hippocampus provide evidence for noradrenergic-glucocorticoid interaction in emotional memory retrieval . Gene polymorphisms of endothelial nitric oxide synthase enzyme associated with pulmonary hypertension in patients with P48444 . In this cross-sectional controlled study , we aimed to investigate the role of polymorphisms of the angiotensin-converting enzyme ( P12821 ) and endothelial nitric oxide synthase ( P29474 ) genes on pulmonary hypertension ( PH ) in patients with chronic obstructive pulmonary disease ( P48444 ) . Forty-two ( 41 male , 1 female , mean age : 62 +/- 7 years ) P48444 patients and 40 ( all male , mean age : 60 +/- 8 years ) healthy controls were included . Respiratory function tests , arterial blood gases , and echocardiographic examinations were performed . P12821 and P29474 genotypes were determined using PCR . The P12821 and P29474 genotype distribution was not significantly different between P48444 patients and controls . On comparing pulmonary artery pressures in different P29474 genotypes , the mean pulmonary artery pressure ( Ppa ) in patients with the BB genotype was significantly higher than in patients with the nonBB genotypes ( 41.3 +/- 17.7 mmHg vs. 27.3 +/- 11.2 mmHg , P = 0.02 ) . However , there was no difference in P12821 genotype distributions between P48444 patients with and without pulmonary hypertension . In stepwise linear regression analysis for predicting pulmonary artery pressure , PaO2 and polymorphism of P29474 gene were found to be independent variables . In conclusion , BB-type polymorphism of the P29474 gene has been associated with PH in addition to hypoxemia . However , P12821 gene polymorphism was not found to be associated with PH . Adrenergic receptor polymorphisms associated with resting heart rate : the HyperGEN Study . The association between polymorphisms in the beta1 , beta2 and alpha2B adrenergic receptor ( ADR ) genes ( P08588 , P07550 and P18089 ) and resting heart rate was examined in white and African-American participants of the HyperGEN Study . All analyses were adjusted for age , sex , body mass index , alcohol use , smoking status and daily exercise within strata of race , hypertension status and beta-blocker use . The Ser49Gly polymorphism of the beta1 ADR was associated with resting heart rate in hypertensive African-Americans and hypertensive whites taking beta-blockers , with carriers of the DB00145 allele having a higher mean resting heart rate by 2.7 and 4.4 beats per minute ( bpm ) , respectively . The Arg389Gly polymorphism of the beta1 ADR was associated with lower heart rate in the normotensive African-American sample . A beta1 haplotype ( Ser49Gly-Arg389Gly ) was modestly associated with resting heart rate in the hypertensive African-Americans . The alpha2B C/A polymorphism was associated with heart rate in hypertensive whites , and both whites and African-Americans taking beta-blockers , with carriers of the A allele having a higher mean resting heart rate . In summary , each of the ADR gene polymorphisms was associated with heart rate in at least one stratum studied , but there was no consistent association from which one would infer a large genetic contribution to heart rate . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Porphyromonas gingivalis-nucleoside-diphosphate-kinase inhibits DB00171 -induced reactive-oxygen-species via Q99572 receptor/NADPH-oxidase signalling and contributes to persistence . Ligation of Q99572 receptors with a ' danger signal ' , extracellular DB00171 ( eATP ) , has recently been shown to result in production of intracellular reactive-oxygen-species ( ROS ) in macrophages . We show that primary gingival epithelial cells ( GECs ) produce sustained , robust cellular ROS upon stimulation by eATP . The induction of ROS was mediated by Q99572 receptor signalling coupled with NADPH-oxidase activation , as determined by pharmacological inhibition and RNA interference . Furthermore , Porphyromonas gingivalis , an oral opportunistic pathogen , upregulated the antioxidant glutathione response , modulated eATP-induced cytosolic and mitochondrial ROS generated through Q99572 /NADPH-oxidase interactome , and subsequently blocked oxidative stress in GECs via temporal secretion of a P. gingivalis effector , nucleoside-diphosphate-kinase ( Ndk ) . An ndk-deficient P. gingivalis mutant lacked the ability to inhibit ROS production and persist intracellularly following eATP stimulation . Treatment with recombinant Ndk significantly diminished eATP-evoked ROS production . P. gingivalis infection elicited a strong , time-dependent increase in anti-oxidativemitochondrial P55851 levels , whereas ndk-deficient mutant did not cause any change . The results reveal a novel signalling cascade that is tightly coupled with eATP signalling and ROS regulation . Ndk by P. gingivalis counteracts these antimicrobial signalling activities by secreting Ndk , thus contributing to successful persistence of the pathogen . Correlations between genetic variance and adiposity measures , and gene x gene interactions for obesity in postmenopausal Vietnamese women . Although environmental factors are important , there is considerable evidence that genes also have a significant role in the pathogenesis of obesity . We conducted a population-based study to investigate the relationship between candidate genes for obesity ( P25874 , P55851 , P18089 , P13945 , LEPR , P11473 and P03372 ) and adiposity measures ( body mass index , body fat percentage , weight , waist circumference and waist-hip ratio ) in terms of individual gene and gene x gene interaction in models unadjusted and adjusted for covariates ( age , years since menopause , educational level and total energy intake ) . Postmenopausal women with TC genotype of P03372 gene had higher body fat percentage than those with TT genotype in the models unadjusted and adjusted for the covariates ( P = 0.006 in adjusted model ) . In multiple logistic regression analysis , BsmI and ApaI SNPs of P11473 genes were significantly associated with overweight and obesity . The P55851 - P11473 ApaI interaction to susceptibility of overweight and obesity was first observed from logistic regression analysis , and then confirmed in the multifactor dimensionality reduction method unadjusted and adjusted for the covariates . This interaction had 69.09 % prediction accuracy for overweight and obesity ( P = 0.001 , sign test ) . In conclusion , the study suggests the significant association of P03372 and P11473 genes with adiposity measures and the P55851 - P11473 ApaI interaction to susceptibility to being overweight and obesity in postmenopausal Vietnamese women . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Deliberate self-harm is associated with allelic variation in the tryptophan hydroxylase gene ( P17752 A779C ) , but not with polymorphisms in five other serotonergic genes . BACKGROUND : There is a heritable component to suicidal behaviour , encouraging the search for the associated risk alleles . Given the putative role of the 5-HT ( 5-hydroxytryptamine ; serotonin ) system in suicidal behaviour , serotonergic genes are leading candidates . In particular , several studies have reported an association with variants in the tryptophan hydroxylase ( P17752 ) gene . METHOD : We studied six serotonergic gene polymorphisms in a well-characterized sample of 129 deliberate self-harm subjects and 329 comparison subjects . The polymorphisms were P17752 ( A779C ) , 5-HT transporter ( 5-HTT , LPR S/L ) , monoamine oxidase A ( P21397 G941T ) , P28222 receptor ( P28222 G861C ) , 5- Q13049 receptor ( P28223 T102C ) , and P28335 receptor ( P28335 Cys23Ser ) . Genotyping was done using polymerase chain reaction ( PCR ) -based assays . The primary analyses compared allele and genotype frequencies between cases and controls . There were a limited number of planned secondary analyses within the deliberate self-harm group . RESULTS : The P17752 A779 allele was more common in deliberate self-harm subjects than in controls ( OR 1.38 , 95 % CI 1.02-1.88 ; P = 0.03 ) . None of the other polymorphisms was associated with deliberate self-harm . Within the deliberate self-harm group there were no associations with impulsivity , suicide risk , lifetime history of depression , or family history of deliberate self-harm . CONCLUSIONS : Our data extend the evidence that allelic variation in the P17752 gene is a risk factor for deliberate self-harm . No evidence was found to implicate the other polymorphisms . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . Radioimmunoassay method for baboon plasma gonadotropins . Double antibody radioimmunoassay methods were developed for the determination of baboon luteinizing hormone ( bLH ) and follicle stimulating hormone ( bFSH ) . The bLH radioimmunoassay employs a unique anti-ovine LH serum ( P07093 -15 ) and ovine LH ( LER-1056- P06681 ) for radioiodination , while the bFSH radioimmunoassay employs an heterologous system , i.e. , an anti-ovine DB00094 serum ( H-31 ) and purified human DB00094 for radioiodination . The reference standard used in both assays is a crude rhesus pituitary extract ( LER-1909-2 ) . Elevated endogeneous baboon plasma DB00024 and prolactin induced by the intravenous administration of 500 microgram of TRH had no influence on the levels of LH and DB00094 , whereas simultaneous intravenous administration of 100 microgram P01148 and 500 microgram TRH raised the levels of LH and DB00094 in plasma . hPRL , hCG , hTSH and hGH did not cross react with either the bLH or the bFSH assay system . The determination of plasma LH and DB00094 concentrations in daily samples from 6 mature female baboons throughout ovulatory menstrual cycles revealed patterns qualitatively similar to those of the rhesus monkey and human females . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients .

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interacts_with DB00641?

multiple_choice

Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization . Determination of the type and quantity of sialic acid in the egg jelly coat of the sea urchin Paracentrotus lividus using capillary LC- P19957 -MS/MS . Sialic acid is a terminal sugar of carbohydrate chains that participates in numerous biological events . Recent studies have explored the mechanism of carbohydrate-mediated fertilisation to understand the biochemistry of fertilisation , although the type and quantity of sialic acid and the role of sialic acid during fertilisation remain unknown . Echinoderm fertilisation in particular has been studied extensively , yet our understanding of the mechanisms of carbohydrate-mediated fertilisation and the role of sialic acid remains incomplete . In this study , we characterised the sialic acid types in the egg jelly coat of the sea urchin , Paracentrotus lividus , using the sensitive analytical system capillary liquid chromatography electro-spray ionisation tandem mass spectrometry ( capLC- P19957 -MS/MS ) . First , we isolated the egg jelly coat and released its sialic acid using acid treatment . These sialic acids were derivatised with 1,2-diamino-4,5-methylenediaoxy-benzene dihydrochloride ( P28068 ) and injected into the capLC- P19957 -MS/MS system . When compared with standards , we identified twelve different types of sialic acid according to their retention times and collision-induced dissociation fragments . The mass spectral data revealed that Neu5Gc , Neu5Ac , Neu5GcS , and Neu5Gc9Ac were the predominant types of sialic acid in the sea urchin jelly coat , with Neu5Gc being the most abundant . Other types of sialic acid detected included Neu5AcS , Neu5Gc7,9Ac2 , Neu5,9Ac2 , Neu5Gc8Ac , Neu5Gc7Ac , Neu5,7Ac2 , Neu5Gc8,9Ac2 , and Neu5,8Ac2 . The types and quantities of sialic acid that we detected in the egg jelly coat will aid in the discovery of new sialic acid-specific receptors on the sperm membrane . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Metabolic fate of 2,2-dimethylbutyryl moiety of simvastatin in rats : identification of metabolites by gas chromatography/mass spectrometry . Metabolic pathways of simvastatin ( DB00641 ) , a lactone prodrug of an inhibitor of P04035 , were elucidated in male rats , using the [ 14C ] -labelled compound . Evidence has been obtained for hydrolysis of simvastatin and its metabolites at their 2,2-dimethylbutyryl moieties . Metabolites identified in plasma were 2,2-dimethylbutyric acid ( P28068 ) , 2,2-dimethyl-3-hydroxybutyric acid ( DMHB ) and an open chain hydroxy acid of simvastatin : metabolites identified in urine were DMHB , a glucuronide and the glycine conjugate of P28068 . They were characterized by gas chromatography/electron impact and chemical ionization mass spectrometry as phenacyl or pertrimethylsilylated derivatives . The structures of the metabolites and the aglycone of the glucuronide were confirmed as phenacyl esters by comparison of their chromatographic data and mass spectra with those of the phenacyl derivatives of authentic compounds . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . P28067 and - P28068 genes are both required for MHC class II/peptide complex formation in antigen-presenting cells . Major histocompatibility complex ( MHC ) class II molecules are highly polymorphic cell-surface glycoproteins that present antigenic peptides to P01730 + T lymphocytes . The normal assembly of class II molecules with cognate peptides for antigen presentation requires an accessory function provided by a gene mapping to the class II region of the HLA complex . The isolation of somatic cell mutants of antigen-presenting cells ( P25054 ) has shown that at least one gene which maps between HLA-DP and HLA-DQ , provisionally designated c2p-1 ( ref. 3 ) , mediates this process . Here we describe a unique new mutant 2.2.93 , which manifests defective formation of class II/peptide complexes like that described in c2p-1 mutants . We show that ( 1 ) mutant 2.2.93 contains a mutation in P28067 , and a representative c2p-1 mutant , 9.5.3 , contains a mutation in P28068 ; and ( 2 ) transfection and expression of P28067 complementary DNA in 2.2.93 , and P28068 cDNA in 9.5.3 , reverses their mutant phenotypes . These results show that P28067 and - P28068 , genes of previously unknown function mapping between HLA-DP and HLA-DQ , are required for the normal assembly of peptides with MHC class II molecules . They suggest that P28067 and - P28068 encode subunits of a functional heterodimer which is critical in the pathway of class II antigen presentation . Combinations of dominant-negative class II transactivator , p300 or P50750 proteins block the expression of MHC II genes . The class II transactivator ( P33076 ) regulates not only the transcription of HLA-DR , -DQ , -DP , but also invariant chain , P28067 and P28068 genes . A hybrid mutant P33076 protein , which contained residues from positions 302 to 1130 in P33076 fused to the enhanced green fluorescent protein ( EdCIITA ) , inhibited the function of the wild-type protein . EdCIITA extinguished the inducible and constitutive expression of MHC II genes in epithelial cells treated with P01579 and B lymphoblastoid cells respectively . Also , it blocked T cell activation by superantigen . This inhibition correlated with the localization of EdCIITA but not P33076 in the cytoplasm of cells . However , when EdCIITA was co-expressed with a dominant-negative form of the nucleoporin Nup214/ P35658 , it also accumulated in the nucleus . These data suggest that EdCIITA not only competes with the wild-type protein for the binding to MHC II promoters but sequesters a critical co-factor of P33076 in the cytoplasm . P33076 also recruits the histone acetyltransferase DB02527 responsive element binding protein ( CREB ) binding protein and positive transcription elongation factor b ( p-TEFb ) for the transcription of MHC II genes . Dominant-negative p300 ( DNp300 ) or P50750 ( DNCDK9 ) proteins inhibited the function of P33076 and of the P40879 promoter . Thus , combinations of EdCIITA and DNp300 and/or DNCDK9 proteins extinguished the transcription of MHC II genes . They might become useful for future genetic therapeutic approaches in organ transplantation and autoimmune diseases . P00747 activator inhibitor-1 and vitronectin expression level and stoichiometry regulate vascular smooth muscle cell migration through physiological collagen matrices . BACKGROUND : Vascular smooth muscle cell ( VSMC ) migration is a critical process in arterial remodeling . Purified plasminogen activator inhibitor-1 ( P05121 ) is reported to both promote and inhibit VSMC migration on two-dimensional ( D ) surfaces . OBJECTIVE : To determine the effects of P05121 and vitronectin ( VN ) expressed by VSMC themselves on migration through physiological collagen matrices . METHODS : We studied migration of wild-type ( WT ) , P05121 -deficient , VN-deficient , P05121 /VN doubly-deficient ( DKO ) and P05121 -transgenic ( Tg ) VSMC through three-D collagen gels . RESULTS : WT VSMC migrated significantly slower than P05121 - and VN-deficient VSMC , but significantly faster than DKO VSMC . Experiments with recombinant P05121 suggested that basal VSMC P05121 expression inhibits migration by binding VN , which is secreted by VSMC and binds collagen . However , P05121 -over-expressing Tg VSMC migrated faster than WT VSMC . Reconstitution experiments with recombinant P05121 mutants suggested that the pro-migratory effect of P05121 over-expression required its anti-plasminogen activator ( PA ) and P01130 -related protein ( Q14764 ) binding functions , but not VN binding . While promoting VSMC migration in the absence of P05121 , VN inhibited the pro-migratory effect of active P05121 . CONCLUSIONS : In isolation , VN and P05121 are each pro-migratory . However , via formation of a high-affinity , non-motogenic complex , P05121 and VN each buffers the other 's pro-migratory effect . The level of P05121 expression by VSMC and the concentration of VN in extracellular matrix are critical determinants of whether P05121 and VN promote or inhibit migration . These findings help to rectify previously conflicting reports and suggest that P05121 /VN stoichiometry plays an important role in VSMC migration and vascular remodeling . Determination of free N-acetylneuraminic acid in human body fluids by high-performance liquid chromatography with fluorimetric detection . Determinations of both the free and bound form of N-acetyl-neuraminic acid ( NANA ) in several human body fluids , such as serum , cerebrospinal fluid ( P04141 ) , saliva , urine , amniotic fluid , and milk were carried out by HPLC with fluorimetric detection . The method utilized 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride ( P28068 ) as a fluorimetric derivatizing reagent . Free-form NANA was obtained from the body fluids after ultrafiltration with Microcon 10 ( YM-10 cellulose membrane , filtration limit M(r) = 10,000 , Amicon ) . The P28068 derivative of NANA was separated isocratically by a Nucleosil 5C18 column with a mixture of 0.1 M sodium phosphate buffer ( pH 2.0 ) -methanol ( 75:25 , v/v ) . A gradient elution system was used for urine analysis . Analysis times were 10-30 min . Recoveries of free NANA by ultrafiltration were satisfactory : 95.66 +/- 1.80 % for serum and 97.27 +/- 1.55 % for P04141 , respectively . The high sensitivity and specificity render this method applicable to all the body fluids tested . Although a physiological role for free NANA has not yet been elucidated , the method presented promises to contribute to the basic understanding of the NANA metabolism . Activity , pharmacological inhibition and biological regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Trypanosoma brucei . Activity of hydroxymethylglutaryl-coenzyme A ( HMG- DB01992 ) reductase , the key enzyme in the biosynthesis of steroids and polyisoprenoids in mammalian cells , has been detected in both the bloodstream form and the culture-adapted procyclic form of Trypanosoma brucei ( 3.7 +/- 0.6 and 12.7 +/- 1.8 pmol mevalonate produced min-1 ( mg cell protein ) -1 , respectively ) . The enzyme activity is enriched 6-fold in microsomal fractions . Several competitive inhibitors of mammalian P04035 , including synvinolin ( simvastatin ) , inhibit the multiplication of both forms of trypanosome in vitro ( IC50 , approx. 25-50 microM after 2-3 days ) . This growth inhibition is potentiated by agents interfering with the exogenous supply of cholesterol , such as antibodies blocking the low-density lipoprotein ( LDL ) receptor , or 5 microM chloroquine . Conversely , growth inhibition by synvinolin can be largely reverted either by 300 nM LDL or by products of the mevalonate pathway , such as 20 mM mevalonate and in procyclics by 100 microM squalene or cholesterol . In procyclics , low concentrations of synvinolin selectively inhibit the incorporation of [14C]acetate into sterols , but not into fatty acids . These results argue for a critical role in trypanosomes of a mevalonate pathway , that is involved in the biosynthesis of sterol and probably of other metabolites . The P04035 activity is decreased 2-fold in procyclics incubated with 4 mM mevalonate and increased 2-fold in the presence of 2.5 microM synvinolin . DB00641 also upregulates LDL binding up to 4-fold . These data suggest that P04035 and P01130 expression are regulated in T. brucei as in mammalian cells , to ensure sterol homeostasis . A common haplotype of the nicotine acetylcholine receptor alpha 4 subunit gene is associated with vulnerability to nicotine addiction in men . DB00184 is the major addictive substance in cigarettes , and genes involved in sensing nicotine are logical candidates for vulnerability to nicotine addiction . We studied six single-nucleotide polymorphisms ( SNPs ) in the P43681 gene and four SNPs in the P17787 gene with respect to nicotine dependence in a collection of 901 subjects ( 815 siblings and 86 parents ) from 222 nuclear families with multiple nicotine-addicted siblings . The subjects were assessed for addiction by both the Fagerstrom Test for DB00184 Dependence ( FTND ) and the Revised Tolerance Questionnaire ( RTQ ) . Because only 5.8 % of female offspring were smokers , only male subjects were included in the final analyses ( 621 men from 206 families ) . Univariate ( single-marker ) family-based association tests ( FBATs ) demonstrated that variant alleles at two SNPs , rs1044396 and rs1044397 , in exon 5 of the P43681 gene were significantly associated with a protective effect against nicotine addiction as either a dichotomized trait or a quantitative phenotype ( i.e. , age-adjusted FTND and RTQ scores ) , which was consistent with the results of the global haplotype FBAT . Furthermore , the haplotype-specific FBAT showed a common ( 22.5 % ) P43681 haplotype , GCTATA , which was significantly associated with both a protective effect against nicotine addiction as a dichotomized trait ( Z=-3.04 , P < .005 ) and significant decreases of age-adjusted FTND ( Z=-3.31 , P < .005 ) or RTQ scores ( Z=-2.73 , P=.006 ) . Our findings provide strong evidence suggesting a common P43681 haplotype might be protective against vulnerability to nicotine addiction in men . Effects of serotonin on expression of the P01130 family member Q92673 and 7-ketocholesterol-induced apoptosis in human vascular smooth muscle cells . Serotonin ( 5-HT ) is a known mitogen for vascular smooth muscle cells ( VSMCs ) . The dedifferentiation and proliferation/apoptosis of VSMCs in the arterial intima represent one of the atherosclerotic changes . Q92673 , a member of low-density lipoprotein receptor family , may contribute to the proliferation of VSMCs in neointimal hyperplasia . We conducted an in vitro study to investigate whether 5-HT is involved in Q92673 expression in human VSMCs and apoptosis of VSMCs induced by 7-ketocholesterol ( 7KCHO ) , an oxysterol that destabilizes plaque . 5-HT enhanced the proliferation of VSMCs , and this effect was abolished by sarpogrelate , a selective 5- Q13049 receptor antagonist . Sarpogrelate also inhibited the 5-HT-enhanced Q92673 mRNA expression in VSMCs . Furthermore , 5-HT suppressed the 7KCHO-induced apoptosis of VSMCs via caspase-3/7-dependent pathway . These findings provide new insights on the changes in the differentiation stage of VSMCs mediated by 5-HT . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Rare human nicotinic acetylcholine receptor α4 subunit ( P43681 ) variants affect expression and function of high-affinity nicotinic acetylcholine receptors . DB00184 , the primary psychoactive component in tobacco smoke , produces its behavioral effects through interactions with neuronal nicotinic acetylcholine receptors ( nAChRs ) . α4β2 nAChRs are the most abundant in mammalian brain , and converging evidence shows that this subtype mediates the rewarding and reinforcing effects of nicotine . A number of rare variants in the P43681 gene that encode the α4 nAChR subunit have been identified in human subjects and appear to be underrepresented in a cohort of smokers . We compared three of these variants ( α4R336C , α4P451L , and α4R487Q ) to the common variant to determine their effects on α4β2 nAChR pharmacology . We examined [(3)H]epibatidine binding , interacting proteins , and phosphorylation of the α4 nAChR subunit with liquid chromatography and tandem mass spectrometry ( LC-MS/MS ) in P29320 293 cells and voltage-clamp electrophysiology in Xenopus laevis oocytes . We observed significant effects of the α4 variants on nAChR expression , subcellular distribution , and sensitivity to nicotine-induced receptor upregulation . Proteomic analysis of immunopurified α4β2 nAChRs incorporating the rare variants identified considerable differences in the intracellular interactomes due to these single amino acid substitutions . Electrophysiological characterization in X. laevis oocytes revealed alterations in the functional parameters of activation by nAChR agonists conferred by these α4 rare variants , as well as shifts in receptor function after incubation with nicotine . Taken together , these experiments suggest that genetic variation at P43681 alters the assembly and expression of human α4β2 nAChRs , resulting in receptors that are more sensitive to nicotine exposure than those assembled with the common α4 variant . The changes in nAChR pharmacology could contribute to differences in responses to smoked nicotine in individuals harboring these rare variants . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . In vitro and in vivo efficacy of PEGylated diisopropyl fluorophosphatase ( DFPase ) . Highly toxic organophosphorus compounds that irreversibly inhibit the enzyme acetycholinesterase ( P22303 ) , including nerve agents like tabun , sarin , or soman , still pose a credible threat to civilian populations and military personnel . New therapeutics that can be used as a pretreatment or after poisoning with these compounds , complementing existing treatment schemes such as the use of atropine and P22303 reactivating oximes , are currently the subject of intense research . A prominent role among potential candidates is taken by enzymes that can detoxify nerve agents by hydrolysis . Diisopropyl fluorophosphatase ( DFPase ) from the squid Loligo vulgaris is known to effectively hydrolyze DB00677 and the range of G-type nerve agents including sarin and soman . In the present work , DFPase was PEGylated to increase biological half-life , and to lower or avoid an immunogenic reaction and proteolytic digest . Addition of linear polyethylene glycol ( PEG ) chains was achieved using mPEG- Q6T4R5 esters and conjugates were characterized by electrospray ionization -- time of flight -- mass specrometry ( P19957 -ToF-MS ) . PEGylated wildtype DFPase and a mutant selective for the more toxic stereoisomers of the agents were tested in vivo with rats that were challenged with a subcutaneous 3x LD(50) dose of soman . While wildtype DFPase prevented death only at extremely high doses , the mutant was able keep the animals alive and to minimize or totally avoid symptoms of poisoning . The results serve as a proof of principle that engineered variants of DFPase are potential candidates for in vivo use if substrate affinity can be improved or the turnover rate enhanced to lower the required enzyme dose . Mitogenic signaling of urokinase receptor-deficient kidney fibroblasts : actions of an alternative urokinase receptor and P01130 -related protein . The urokinase receptor ( Q03405 ) attenuates myofibroblast recruitment and fibrosis in the kidney . This study examined the role of Q03405 and its co-receptor P01130 -related protein ( Q14764 ) in the regulation of kidney fibroblast proliferation and extracellular signal-regulated kinase ( P29323 ) signaling . Compared with Q03405 +/+ cells , Q03405 -/- kidney fibroblasts were hyperproliferative . Q03405 -/- fibroblast proliferation was 60 % inhibited by an P29323 kinase inhibitor . Q14764 protein was reduced and extracellular accumulation of urokinase-type plasminogen activator ( uPA ) and plasminogen activator inhibitor type 1 ( P05121 ) proteins were greater in Q03405 -/- cultures . Addition of functional uPA protein or Q14764 antisense RNA significantly increased P29323 signaling and cell mitosis in both genotypes . Enhanced Q03405 -/- fibroblast proliferation was reversed by a recombinant nonfunctional uPA peptide . The density of cell-bound fluor-uPA was similar between Q03405 -/- and Q03405 +/+ fibroblasts ( 78 +/- 6 versus 92 +/- 16 units ) . These data suggest that Q03405 -deficient kidney fibroblasts express lower levels of its scavenger co-receptor Q14764 , resulting in greater extracellular accumulation of uPA and P05121 . Enhanced proliferation of Q03405 -/- fibroblasts seems to be mediated by uPA-dependent P29323 signaling via an alternative urokinase receptor . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . Genetics of late-onset Alzheimer 's disease : update from the alzgene database and analysis of shared pathways . The genetics of late-onset Alzheimer 's disease ( LOAD ) has taken impressive steps forwards in the last few years . To date , more than six-hundred genes have been linked to the disorder . However , only a minority of them are supported by a sufficient level of evidence . This review focused on such genes and analyzed shared biological pathways . Genetic markers were selected from a web-based collection ( Alzgene ) . For each SNP in the database , it was possible to perform a meta-analysis . The quality of studies was assessed using criteria such as size of research samples , heterogeneity across studies , and protection from publication bias . This produced a list of 15 top-rated genes : P02649 , P10909 , Q13492 , Q2M3D2 , O00499 , P17927 , Q92673 , Q13470 , P10145 , P01130 , P01034 , P17787 , Q8WY21 , P01375 , and P41597 . A systematic analysis of gene ontology terms associated with each marker showed that most genes were implicated in cholesterol metabolism , intracellular transport of beta-amyloid precursor , and autophagy of damaged organelles . Moreover , the impact of these genes on complement cascade and cytokine production highlights the role of inflammatory response in AD pathogenesis . Gene-gene and gene-environment interactions are prominent issues in AD genetics , but they are not specifically featured in the Alzgene database . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Obesity and breast cancer : the roles of peroxisome proliferator-activated receptor-γ and plasminogen activator inhibitor-1 . Breast cancer is the most prominent cancer among females in the United States . There are a number of risk factors associated with development of breast cancer , including consumption of a high-fat diet and obesity . P00747 activator inhibitor-1 ( P05121 ) is a cytokine upregulated in obesity whose expression is correlated with a poor prognosis in breast cancer . As a key mediator of adipogenesis and regulator of adipokine production , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) is involved in P05121 expression from adipose tissue . We summarize the current knowledge linking Q07869 -γ and P05121 expression to high-fat diet and obesity in the risk of breast cancer . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Polymorphisms in genes implicated in dopamine , serotonin and noradrenalin metabolism suggest association with cerebrospinal fluid monoamine metabolite concentrations in psychosis . BACKGROUND : Homovanillic acid ( HVA ) , 5-hydroxyindoleacetic acid ( 5-HIAA ) and 3-methoxy-4-hydroxyphenylglycol ( MHPG ) are the major monoamine metabolites in the central nervous system ( CNS ) . Their cerebrospinal fluid ( P04141 ) concentrations , reflecting the monoamine turnover rates in CNS , are partially under genetic influence and have been associated with schizophrenia . We have hypothesized that P04141 monoamine metabolite concentrations represent intermediate steps between single nucleotide polymorphisms ( SNPs ) in genes implicated in monoaminergic pathways and psychosis . METHODS : We have searched for association between 119 SNPs in genes implicated in monoaminergic pathways [ tryptophan hydroxylase 1 ( P17752 ) , Q8IWU9 , tyrosine hydroxylase ( TH ) , P20711 ( DDC ) , dopamine beta-hydroxylase ( P09172 ) , catechol-O-methyltransferase ( P21964 ) , monoamine oxidase A ( P21397 ) and P27338 ] and monoamine metabolite concentrations in P04141 in 74 patients with psychotic disorder . RESULTS : There were 42 nominally significant associations between SNPs and P04141 monoamine metabolite concentrations , which exceeded the expected number ( 20 ) of nominal associations given the total number of tests performed . The strongest association ( p = 0.0004 ) was found between P27338 rs5905512 , a SNP previously reported to be associated with schizophrenia in men , and MHPG concentrations in men with psychotic disorder . Further analyses in 111 healthy individuals revealed that 41 of the 42 nominal associations were restricted to patients with psychosis and were absent in healthy controls . CONCLUSIONS : The present study suggests that altered monoamine turnover rates in CNS reflect intermediate steps in the associations between SNPs and psychosis . Determination of MK-0767 enantiomers in human plasma by normal phase LC-MS/MS . A sensitive and selective analytical method for the enantioselective determination of MK-0767 , a dual peroxisome proliferator-activated receptor ( Q07869 ) alpha/gamma agonist , in human plasma has been developed and validated . The chromatography is based on normal-phase chiral separation on a Kromasil , 5 microm , CHI- P28068 250 mm x 4.6 mm column . The detection involves the direct introduction of the normal phase eluent into MS/MS without the addition of a post-column reagent . Atmospheric pressure chemical ionization ( APcI ) mode was selected as the ion source in this method . With proper sample handling and processing procedures , ex vivo interconversion of the enantiomers was kept to minimum during sample collection , preparation and short term storage of frozen human plasma samples . The method was successfully utilized to determine the concentrations of MK-0767 enantiomers in human plasma to support pharmacokinetic investigation in man . The role of de novo ceramide synthesis in the mechanism of action of the tricyclic xanthate D609 . The cytotoxic effects of several chemotherapeutic drugs have been linked to elevated de novo ceramide biosynthesis . However , the relationship between the intracellular site(s) of ceramide accumulation and cytotoxicity is poorly understood . Here we examined the relationship between the site of ceramide deposition and inhibition of protein translation and induction of apoptosis by the antitumor/antiviral xanthate , D609 . In Chinese hamster ovary ( CHO ) - P04264 , P29320 -293 , and NIH-3T3 cells , D609 caused rapid ( 1-5 min ) and sustained eukaryotic initiation factor 2alpha ( eIF2alpha ) phosphorylation followed by apoptosis after 24 h . Concurrently , D609 stimulated de novo ceramide synthesis and increased ceramide mass 2-fold by 2 h in CHO- P04264 cells . In D609-treated CHO- P04264 cells , sphingomyelin synthesis was stimulated by brefeldin A , and P01031 - P28068 -ceramide transport to the Golgi apparatus was blocked , indicating ceramide accumulation in the endoplasmic reticulum ( ER ) . However , D609-mediated eIF2alpha phosphorylation , inhibition of protein synthesis , and apoptosis in CHO- P04264 cells were not attenuated by fumonisin B1 or l-cycloserine . Interestingly , short-chain ceramide promoted eIF2alpha phosphorylation and inhibited protein synthesis in CHO- P04264 cells , indicating that the effectiveness of endogenous ceramide could be limited by access to signaling pathways . Thus , expansion of the ER ceramide pool by D609 was not implicated in early ( eIF2alpha phosphorylation ) or late ( apoptotic ) cytotoxic events . Q9Y5Q5 mutations K317E and S472G from preeclamptic patients alter zymogen activation and cell surface targeting. [ Corrected ] . Q9Y5Q5 is a membrane-bound serine protease that acts as the atrial natriuretic peptide ( P01160 ) convertase in the heart . Recent studies show that corin also activates P01160 in the pregnant uterus to promote spiral artery remodeling and prevent pregnancy-induced hypertension . Two Q9Y5Q5 gene mutations , K317E and S472G , were identified in preeclamptic patients and shown to have reduced activity in vitro . In this study , we carried out molecular modeling and biochemical experiments to understand how these mutations impair corin function . By molecular modeling , the mutation K317E was predicted to alter corin P01130 -2 module conformation . Western blot analysis of K317E mutant in HEK293 cells showed that the mutation did not block corin expression on the cell surface but inhibited corin zymogen activation . In contrast , the mutation S472G was predicted to abolish a β-sheet critical for corin frizzled-2 module structure . In Western blot analysis and flow cytometry , S472G mutant was not detected on the cell surface in transfected HEK293 cells . By immunostaining , the S472G mutant was found in the ER , indicating that the mutation S472G disrupted the β-sheet , causing corin misfolding and ER retention . Thus , these results show that mutations in the Q9Y5Q5 gene may impair corin function by entirely different mechanisms . Together , our data provide important insights into the molecular basis underlying corin mutations that may contribute to preeclampsia in patients . P37268 inhibitors suppress triglyceride biosynthesis through the farnesol pathway in rat hepatocytes . We recently demonstrated that squalene synthase ( P37268 ) inhibitors reduce plasma triglyceride through an P01130 -independent mechanism in Watanabe heritable hyperlipidemic rabbits ( Hiyoshi et al. 2001. Eur. J. Pharmacol. 431 : 345-352 ) . The present study deals with the mechanism of the inhibition of triglyceride biosynthesis by the P37268 inhibitors ER-27856 and RPR-107393 in rat primary cultured hepatocytes . DB01076 , an P04035 inhibitor , had no effect on triglyceride biosynthesis , but reversed the inhibitory effect of the P37268 inhibitors . A squalene epoxidase inhibitor , NB-598 , affected neither triglyceride biosynthesis nor its inhibition by ER-27856 and RPR-107393 . The reduction of triglyceride biosynthesis by ER-27856 and RPR-107393 was potentiated by mevalonolactone supplementation . Treatment of hepatocytes with farnesol and its derivatives reduced triglyceride biosynthesis . In addition , we found that ER-27856 and RPR-107393 significantly reduced the incorporation of [1-(14)C]acetic acid into oleic acid , but not the incorporation of [1-(14)C]oleic acid into triglyceride . Though ER-27856 and RPR-107393 increased mitochondrial fatty acid beta-oxidation , the inhibition of beta-oxidation by RS-etomoxir had little effect on their inhibition of triglyceride biosynthesis . These results suggest that P37268 inhibitors reduce triglyceride biosynthesis by suppressing fatty acid biosynthesis via an increase in intracellular farnesol and its derivatives . Functional alterations in endothelial NO , PGI₂ and EDHF pathways in aorta in ApoE/ P01130 -/- mice . Adequate endothelial production of nitric oxide ( NO ) , endothelium-derived hyperpolarizing factor ( EDHF ) , and prostacyclin ( PGI₂ ) is critical to the maintenance of vascular homeostasis . However , it is not clear whether alterations in each of these vasodilatory pathways contribute to the impaired endothelial function in murine atherosclerosis . In the present study , we analyze the alterations in NO- , EDHF- and PGI₂-dependent endothelial function in the thoracic aorta in relation to the development of atherosclerotic plaques in apoE/LDLR⁻/⁻ mice . We found that in the aorta of 2-month-old apoE/LDLR⁻/⁻ mice there was no lipid deposition , subendothelial macrophage accumulation ; and matrix metalloproteinase ( MMP ) activity was low , consistent with the absence of atherosclerotic plaques . Interestingly , at this stage the endothelium was already activated and hypertrophic as evidenced by electron microscopy , while acetylcholine-induced NO-dependent relaxation in the thoracic aorta was impaired , with concomitant upregulation of cyclooxygenase-2 ( P35354 ) /PGI₂ and EDHF ( epoxyeicosatrienoic acids , EETs ) pathways . In the aorta of 3-6-month-old apoE/LDLR⁻/⁻ mice , lipid deposition , macrophage accumulation and MMP activity in the intima were gradually increased , while impairment of NO-dependent function and compensatory upregulation of P35354 /PGI₂ and EDHF pathways were more accentuated . These results suggest that impairment of NO-dependent relaxation precedes the development of atherosclerosis in the aorta and early upregulation of P35354 /PGI₂ and EDHF pathways may compensate for the loss of the biological activity of NO . Essential role for the ( hepatic ) P01130 in macrophage apolipoprotein E-induced reduction in serum cholesterol levels and atherosclerosis . P02649 ( apoE ) is a high affinity ligand for several receptor systems in the liver , including the low-density lipoprotein ( LDL ) receptor , and non- P01130 sites , like the P01130 -related protein ( Q14764 ) , the putative remnant receptor and/or proteoglycans . Although the liver is the major source of apoE synthesis , apoE is also produced by a wide variety of other cell types , including macrophages . In the present study , the role of the P01130 in the removal of lipoprotein remnants , enriched with macrophage-derived apoE from the circulation , was determined using the technique of bone marrow transplantation ( BMT ) . Reconstitution of macrophage apoE production in apoE-deficient mice resulted in a serum apoE concentration of only 2 % of the concentration in wild-type C57Bl/6 mice . This low level of apoE nevertheless reduced VLDL and LDL cholesterol 12-fold ( P < 0.001 ) and fourfold ( P < 0.001 ) , respectively , thereby reducing serum cholesterol levels and the susceptibility to atherosclerosis . In contrast , reconstitution of macrophage apoE synthesis in mice lacking both apoE and the P01130 induced only a twofold ( P < 0.001 ) reduction in VLDL cholesterol and had no significant effect on atherosclerotic lesion development , although serum apoE levels were 93 % of the concentration in normal C57Bl/6 mice . In conclusion , a functional ( hepatic ) P01130 is essential for the efficient removal of macrophage apoE-enriched lipoprotein remnants from the circulation and thus for normalization of serum cholesterol levels and protection against atherosclerotic lesion development in apoE-deficient mice .

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interacts_with DB00216?

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MnSOD drives neuroendocrine differentiation , androgen independence , and cell survival in prostate cancer cells . An increase in neuroendocrine ( NE ) cell number has been associated with progression of prostate tumor , one of the most frequent cancers among Western males . We previously reported that mitochondrial manganese superoxide dismutase ( MnSOD ) increases during the NE differentiation process . The goal of this study was to find whether MnSOD up-regulation is enough to induce NE differentiation . Several human prostate cancer LNCaP cell clones stably overexpressing MnSOD were characterized and two were selected ( MnSOD-S4 and MnSOD- P28222 ) . MnSOD overexpression induces NE morphological features as well as coexpression of the NE marker synaptophysin . Both MnSOD clones exhibit lower superoxide levels and higher H(2)O(2) levels . MnSOD-overexpressing cells show higher proliferation rates in complete medium , but in steroid-free medium MnSOD- P28222 cells are still capable of proliferation . MnSOD up-regulation decreases androgen receptor and prevents its nuclear translocation . MnSOD also induces up-regulation of Bcl-2 and prevents docetaxel- , etoposide- , or P01375 -induced cell death . Finally , MnSOD-overexpressing cells enhance growth of androgen-independent PC-3 cells but reduce growth of androgen-dependent cells . These results indicate that redox modulation caused by MnSOD overexpression explains most NE-like features , including morphological changes , NE marker expression , androgen independence , inhibition of apoptosis , and enhancement of cell growth . Many of these events can be associated with the androgen dependent-independent transition during prostate cancer progression . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Efficacy , safety and tolerability of oral eletriptan in the acute treatment of migraine : results of a phase III , multicentre , placebo-controlled study across three attacks . The efficacy , safety and tolerability of the P28222 /D receptor agonist eletriptan ( 40 mg and 80 mg ) in acute treatment of migraine was evaluated in a multinational , randomized , double-blind , parallel-group , placebo-controlled , three-attack study treating 1153 patients . In the initial attack , significantly more eletriptan patients reported headache relief and complete pain relief at 2 h vs. placebo ( 40 mg 62 % and 32 % , 80 mg 65 % and 34 % , placebo 19 % and 3 % ; P < 0.0001 ) . Headache relief occurred faster after eletriptan , with more patients at both doses reporting relief 30 min ( P < 0.01 ) and 1 h ( P < 0.0001 ) after treatment than after placebo . There was a significantly lower recurrence rate with eletriptan 80 mg compared with placebo ( P < 0.01 ) . Adverse events for all treatments were generally mild or moderate and self-limiting . DB00216 40 mg and eletriptan 80 mg both appear to be effective and well-tolerated acute migraine treatments . Characterisation of the contractile activity of eletriptan at the canine vascular P28222 receptor . The functional activity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] - 1 H-indole ) at the contractile serotonin ( 5-hydroxytryptamine ; 5-HT ) ' 1B-like ' receptor in dog isolated saphenous vein and basilar artery was investigated . DB00216 , like 5-HT and sumatriptan potently contracted saphenous vein ( pEC50 : 6.3 , 6.9 and 6.1 , respectively ) and basilar artery ( pEC50 7.2 , 7.5 and 6.8 , respectively ) . The maximum responses evoked by eletriptan was , unlike sumatriptan , significantly lower than that to 5-HT ( intrinsic activity saphenous vein : eletriptan 0.57 , 5-HT 1.0 , sumatriptan 0.85 ; basilar artery : eletriptan 0.77 , 5-HT 0.98 , sumatriptan 0.89 ) . Contractions evoked by eletriptan were antagonised by the P28222 /1D receptor antagonist GR125743 ( N- [ 4-methoxy-3- ( 4-methyl piperazin-1-yl ) phenyl ] -3-methyl-4-(4-pyridyl)benzamide ) with pA2 values of 9.1 in saphenous vein and 9.4 in basilar artery . Affinity estimates ( pKA ) for 5-HT and sumatriptan determined from receptor alkylation studies in saphenous vein were 6.6 and 6.3 , respectively , compared to the apparent equilibrium dissociation constant ( pKp ) for eletriptan of 6.8 . The rank order of relative intrinsic efficacies ( epsilon ) was 5-HT > sumatriptan > eletriptan . Thus , eletriptan required greater receptor occupancy ( 4.4-fold ) to evoke an equivalent contraction to 5-HT and sumatriptan in dog isolated saphenous vein . These data demonstrate that eletriptan is a potent partial agonist at the canine vascular P28222 receptor . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . Deliberate self-harm is associated with allelic variation in the tryptophan hydroxylase gene ( P17752 A779C ) , but not with polymorphisms in five other serotonergic genes . BACKGROUND : There is a heritable component to suicidal behaviour , encouraging the search for the associated risk alleles . Given the putative role of the 5-HT ( 5-hydroxytryptamine ; serotonin ) system in suicidal behaviour , serotonergic genes are leading candidates . In particular , several studies have reported an association with variants in the tryptophan hydroxylase ( P17752 ) gene . METHOD : We studied six serotonergic gene polymorphisms in a well-characterized sample of 129 deliberate self-harm subjects and 329 comparison subjects . The polymorphisms were P17752 ( A779C ) , 5-HT transporter ( 5-HTT , LPR S/L ) , monoamine oxidase A ( P21397 G941T ) , P28222 receptor ( P28222 G861C ) , 5- Q13049 receptor ( P28223 T102C ) , and P28335 receptor ( P28335 Cys23Ser ) . Genotyping was done using polymerase chain reaction ( PCR ) -based assays . The primary analyses compared allele and genotype frequencies between cases and controls . There were a limited number of planned secondary analyses within the deliberate self-harm group . RESULTS : The P17752 A779 allele was more common in deliberate self-harm subjects than in controls ( OR 1.38 , 95 % CI 1.02-1.88 ; P = 0.03 ) . None of the other polymorphisms was associated with deliberate self-harm . Within the deliberate self-harm group there were no associations with impulsivity , suicide risk , lifetime history of depression , or family history of deliberate self-harm . CONCLUSIONS : Our data extend the evidence that allelic variation in the P17752 gene is a risk factor for deliberate self-harm . No evidence was found to implicate the other polymorphisms . Characterisation of the 5-HT receptor binding profile of eletriptan and kinetics of [3H]eletriptan binding at human P28222 and P28221 receptors . The affinity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] -1H-indole ) for a range of 5-HT receptors was compared to values obtained for other P28222 /1D receptor agonists known to be effective in the treatment of migraine . DB00216 , like sumatriptan , zolmitriptan , naratriptan and rizatriptan had highest affinity for the human P28222 , P28221 and putative 5-ht1f receptor . Kinetic studies comparing the binding of [3H]eletriptan and [3H]sumatriptan to the human recombinant P28222 and P28221 receptors expressed in HeLa cells revealed that both radioligands bound with high specificity ( > 90 % ) and reached equilibrium within 10-15 min . However , [3H]eletriptan had over 6-fold higher affinity than [3H]sumatriptan at the P28221 receptor ( K(D) ) : 0.92 and 6.58 nM , respectively ) and over 3-fold higher affinity than [3H]sumatriptan at the P28222 receptor ( K(D) : 3.14 and 11.07 nM , respectively ) . Association and dissociation rates for both radioligands could only be accurately determined at the P28221 receptor and then only at 4 degrees C . At this temperature , [3H]eletriptan had a significantly ( P < 0.05 ) faster association rate ( K(on) 0.249 min(-1) nM(-1) ) than [3H]sumatriptan ( K(on) 0.024 min(-1) nM(-1) ) and a significantly ( P < 0.05 ) slower off-rate ( K(off) 0.027 min(-1) compared to 0.037 min(-1) for [3H]sumatriptan ) . These data indicate that eletriptan is a potent ligand at the human P28222 , P28221 , and 5-ht1f receptors and are consistent with its potent vasoconstrictor activity and use as a drug for the acute treatment of migraine headache . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Effects of eletriptan on the peptidergic innervation of the cerebral dura mater and trigeminal ganglion , and on the expression of c-fos and c-jun in the trigeminal complex of the rat in an experimental migraine model . Nociceptive axons and terminals in the supratentorial cerebral dura mater display an intense calcitonin gene-related peptide ( P80511 ) immunoreactivity . In an experimental migraine model , it has been shown that electrical stimulation of the rat trigeminal ganglion induced an increase in the lengths of P80511 -immunoreactive axons , increased size and number of pleomorphic axonal varicosities in the dura mater , and an increased number of c-jun and c-fos protein-expressing nerve cells in the trigeminal complex . We demonstrate the effect of the highly specific and moderately lipophilic serotonin agonist eletriptan ( Pfizer ) which prevents the effects of electrical stimulation in the dura mater . DB00216 also affected the caudal trigeminal complex ; it markedly reduced the numbers of the oncoprotein-expressing cells , mainly after stimulation and to some extent also in nonstimulated animals . DB00216 also affected expression of P80511 in perikarya of trigeminal ganglion cells , insofar as the number of small nerve cells exhibiting a compact P80511 immunoreaction was decreased to one quarter of the original value . In all these respects , eletriptan acted in a similar way to sumatriptan , with the notable exception that eletriptan also blocked the stimulation-induced effects in the nucleus caudalis trigemini and the upper cervical spinal cord ( trigeminal complex ) , whereas sumatriptan did not . It is concluded that eletriptan , acting on perikarya and both the peripheral and the central axon terminals of primary sensory neurons , exerts its antimigraine effect by an agonist action on P28222 /1D receptors throughout the entire trigeminal system , probably by passing the blood-brain-barrier because of its lipophilic character . DB01393 induces plasminogen activator inhibitor-1 gene expression in a O15516 -dependent circadian manner . A functional interaction between peroxisome proliferator-activated receptor alpha ( PPARalpha ) and components of the circadian clock has been suggested , but whether these transcriptional factors interact to regulate the expression of their target genes remains obscure . Here we used a PPARalpha ligand , bezafibrate , to search for PPARalpha-regulated genes that are expressed in a O15516 -dependent circadian manner . Microarray analyses using hepatic RNA isolated from bezafibrate treated-wild type , Clock mutant ( Clk/Clk ) , and PPARalpha-null mice revealed that 136 genes are transcriptionally regulated by PPARalpha in a O15516 -dependent manner . Among them , we focused on the plasminogen activator inhibitor-1 ( P05121 ) gene , because its expression typically shows circadian variation , and it has transcriptional response elements for both Q07869 and O15516 . The bezafibrate-induced expression of P05121 mRNA was attenuated in Clk/Clk mice and in PPARalpha-null mice . The protein levels of PPARalpha were reduced in Clk/Clk hepatocytes . However , the overexpression of PPARalpha could not rescue bezafibrate-induced P05121 expression in Clk/Clk hepatocytes , suggesting that impaired bezafibrate-induced P05121 expression in Clk/Clk mice is not due to reduced PPARalpha expression . Luciferase reporter and chromatin immunoprecipitation analyses using primary hepatocytes demonstrated that DNA binding of both PPARalpha and O15516 is essential for bezafibrate-induced P05121 gene expression . Pull-down assays in vitro showed that both PPARalpha and its heterodimerized partner retinoic acid receptor-alpha can serve as potential interaction targets of O15516 . The present findings revealed that molecular interaction between the circadian clock and the lipid metabolism regulator affects the bezafibrate-induced gene expression . 5-hydroxytryptamine stimulates phosphorylation of Q8TCB0 / Q8NFH3 mitogen-activated protein kinase activation in bovine aortic endothelial cell cultures . 5-Hydroxytryptamine ( 5-HT ) is sequestered and released by endothelial cells , acts as an endothelial cell mitogen , promotes the release of nitric oxide ( NO ) , and has been associated with the Q8TCB0 / Q8NFH3 mitogen-activated protein kinase ( MAPK ) cascade . NO also acts as a cell mitogen and promotes signals that culminate in the phosphorylation of MAPK . The aim of this study was to test whether endothelial 5-HT receptors stimulate dual ( tyrosyl- and threonyl- ) phosphorylation of MAPK through a mitogen-activated protein kinase kinase-1 ( MEK-1 ) and P29474 -dependent pathway in bovine aortic endothelial cells ( BAECs ) . As shown by Western blot analysis , 5-HT and the P28222 -selective agonist 5-nonyloxytryptamine ( 5-NOT ) stimulate time- and concentration-dependent ( 0.001-10 microM ) phosphorylation of MAPK in these cells . The agonist-stimulated phosphorylation of MAPK was blocked by the 5-HT1b-receptor antagonist isamoltane ( 0.01-10 p3M ) and the MEK-1 inhibitor PD 098059 ( [ 2-(2'-amino-3'-methoxy-phenyl)-oxanaphthalen-4-one ] ; 0.01-10 microM¿ . The P29474 inhibitor L-N(omega)-iminoethyl-L-ornithine ( L-NIO ; 0.01-10 microM ) failed to block the 1 microM 5-NOT-stimulated responses . Our findings suggest that the 5-HT receptors ( specifically P28222 ) mediate signals to MEK-1 and subsequently to MAPK through an P29474 -independent pathway in BAECs . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . [ Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture ] . In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10(-7)-10(-10) M it protected cells of sympathetic ganglia , neocortex and continues cell lines under damaging actions of H2O2 ( 0.0001 M ) , NH4CI ( 0.01 M ) and cooling . DB00086 essentially influenced the mode of damaging effect of DB00171 ( 0.001 M ) . Even a short-term exposition ( 20 min ) of PC12 cells with both proteins ( each in the concentration 10(-9) M ) led to sharp alterations in intracellular DB00171 - or Ca(2+)-activated proteolysis . In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation , improvement of cell adhesion , high survival rate , the increase in quantity and length of processes and their arborisation . Electronic microscopy established the character of structural rearrangements of nervous tissue cells ( neurons , astrocytes , oligodendrocytes ) , reflecting the protective action of plasminogen and streptokinase . In the presence of plasminogen and especially streptokinase , the total number of cultured glioma P13671 and neuroblastoma IMR-32 cells , the intracellular contents of protein , RNA and DNA increased several-fold . Addition of plasminogen promoted formation of processes by neuroblastoma cells , this suggests initiation of differentiation of cellular elements . In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells . These proteins did not cause transformation of PC12 enterochromaffine cells to neurons , though plasminogen facilitated it . P00747 addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium , and streptokinase did not lose its plasminogen-activating capacity . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . High content analysis of human fibroblast cell cultures after exposure to space radiation . Space travel imposes risks to human health , in large part by the increased radiation levels compared to those on Earth . To understand the effects of space radiation on humans , it is important to determine the underlying cellular mechanisms . While general dosimetry describes average radiation levels accurately , it says little about the actual physiological impact and does not provide biological information about individual cellular events . In addition , there is no information about the nature and magnitude of a systemic response through extra- and intercellular communication . To assess the stress response in human fibroblasts that were sent into space with the Foton-M3 mission , we have developed a pluralistic setup to measure DNA damage and inflammation response by combining global and local dosimetry , image cytometry and multiplex array technology , thereby maximizing the scientific output . We were able to demonstrate a significant increase in DNA double-strand breaks , determined by a twofold increase of the gamma- P16104 signal at the level of the single cell and a threefold up-regulation of the soluble signal proteins P13501 , P05231 , P10145 , beta-2 microglobulin and P80511 , which are key players in the process of inflammation , in the growth medium . Porcine carotid vascular effects of eletriptan ( UK-116,044 ) : a new P28222 /1D receptor agonist with anti-migraine activity . It has been suggested that opening of cephalic arteriovenous anastomoses may be involved in the headache phase of migraine . Indeed , a number of acutely acting anti-migraine drugs , including the ergot alkaloids and sumatriptan , constrict porcine carotid arteriovenous anastomoses . In this study , using pentobarbital anaesthetised pigs , we investigated the effects of eletriptan , a close structural analogue of sumatriptan , on the distribution of common carotid artery blood flow into arteriovenous anastomotic and nutrient ( capillary ) fractions . DB00216 ( 10 , 30 , 100 , 300 and 1000 microg kg(-1) , i.v. ) decreased the total carotid blood flow , exclusively by decreasing cephalic arteriovenous anastomotic blood flow ; nutrient blood flow , particularly to the ear , skin and fat , was significantly increased . The doses of eletriptan needed to reduce arteriovenous anastomotic blood flow and conductance by 50 % ( ED50 ) were , respectively , 117+/-21 microg kg(-1) ( 251+/-45 nmol kg(-1) ) and 184+/-42 microg kg(-1) ( 396+/-91 nmol kg(-1) ) ; the highest dose caused reductions of 84+/-3 % and 77+/-4 % , respectively . The eletriptan-induced changes in carotid haemodynamics were clearly attenuated by pretreating the pigs with the selective P28222 /1D receptor antagonist GR127935 ( 0.5 mg kg(-1) ) . On the basis of these results , we conclude that ( 1 ) the eletriptan-induced constriction of cephalic arteriovenous anastomoses as well as the arteriolar dilatation in head tissues is predominantly mediated by P28222 /1D receptors , and ( 2 ) eletriptan should be effective in aborting migraine headache . Clinical studies have already demonstrated its therapeutic action in migraine patients . DB00216 Pfizer . Pfizer has developed and launched eletriptan , a P28222 /1D agonist , for the potential treatment of migraine with and without aura . DB00216 has 6-fold greater affinity for the P28221 receptor than sumatriptan , and a 3-fold greater affinity for the P28222 receptor [ 249570 ] . DB00216 pharmacology has also been evaluated in vitro in comparison with zolmitriptan ( AstraZeneca plc ) and naratriptan ( GlaxoSmithKline plc ) [ 290116 ] . Biochemical markers and genetic research of ADHD . ADHD ( attention hyperactivity disorder ) is a polygenetic disorder with various candidate genes . At this time , more than thirty dopaminergic , noradrenergic , serotonergic and GABA-ergic genes are known . The research of only some candidate genes ( P21917 , Q01959 , P21918 , P09172 , P31645 , P28222 and P60880 ) brought relatively consistent results confirming the heredity of ADHD syndromes . The results of research of other genes ( P14416 , P35462 , MAO , ADR2A , GABA A3 , GABA B3 ) are not clear yet . This paper summarizes the most important genetic data in correlations with biochemical periphery parameters ( especially for P09172 , HVA , MHPG , serotonin ) . Hypothetically , certain subgroups of ADHD may be identified by correlation of biochemical characteristics and some candidate genes . The paper discusses some implications for future research. Review . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Intramuscular gene transfer of P80511 inhibits neointimal hyperplasia after balloon injury in the rat abdominal aorta . P80511 is a well-known neuropeptide that has various protective effects on cardiovascular system . Our previous studies have shown that P80511 inhibits vascular smooth muscle cell ( VSMC ) proliferation in vitro . The present study aimed to explore the role of the P80511 in neointimal formation after balloon injury in the rat aortic wall and the underlying mechanism . Gene transfer of P80511 was performed with the use of intramuscular electroporation in a balloon-injured rat aorta model . Apoptosis in VSMCs was determined by electrophoresis assessment of DNA fragmentation and terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay . Overexpression of the P80511 gene significantly inhibited the neointimal formation after balloon injury compared with the mock transfer , as assessed by the intima-to-media ratio 14 days after balloon injury ( 29.2 +/- 3.7 % vs. 52.7 +/- 5.4 % ; n = 9-12 , P < 0.05 ) . In addition , P80511 gene expression increased the number of apoptotic cells in the neointima in vivo 14 days after balloon injury . Similarly , the addition of bioactive P80511 and the nitric oxide donor induced similar apoptosis in cultured VSMCs . The antagonist of the P80511 (1) receptor and inhibitors of DB02527 -PKA and nitric oxide blocked P80511 -mediated apoptosis . Furthermore , P80511 gene transfer increased inducible nitric oxide synthase and p53 but decreased P12004 and Bcl-2 protein levels in balloon-injured rat aorta . Our data demonstrated that P80511 potently inhibited neointimal thickening in the rat aorta , at least in part through its distinct effects on apoptosis and proliferation of VSMCs both in vivo and in vitro . Therefore , delivery of the P80511 gene may have therapeutic implications in limiting vascular restenosis . Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor . Mouse pluripotent hematopoietic stem cells ( PHSC ) were fractionated based on size and density using counterflow centrifugal elutriation ( CCE ) . These heterogeneous PHSC populations were further enriched by subtraction of cells with lineage-specific markers ( Lin- ) followed by positive sorting for c-kit expression . The cells were characterized for their functional and biochemical properties . We defined a subpopulation of c-kit-positive cells that expressed high numbers of c-kit receptors ( c-kitBR ) . One hundred c-kitBR cells from either low- or higher-density fractions were sufficient to repopulate the lymphohematopoietic system in WBB6F1-W/Wv ( W/Wv ) recipients , whereas no PHSC were found in cells with low ( c-kitDULL ) or no ( c-kitNEG ) c-kit expression . Lin- c-kitBR cells were separated into RhoDULL and RhoBR subsets based on their ability to efflux rhodamine 123 ( Rho ) . The PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin- c-kitBR RhoBR cells correlated directly with the number of day 12 colony-forming unit-spleen ( CFU- P28222 ) in each fraction . We were not able to enrich further for PHSC using monoclonal antibodies to the cell-surface markers AA4.1 or P01730 , which have been used by others to isolate PHSC . The small , low-density Lin- c-kitBR subset contained PHSC and few CFU- P28222 . This enabled us to assay PHSC for expression of the flk-2 gene , which encodes a tyrosine kinase receptor present on fetal liver PHSC . Purified RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA . We suggest that AA4.1 , P01730 and flk-2 are expressed as stage-specific markers on PHSC in cell cycle . DNA sequence polymorphisms in genes involved in the regulation of dopamine and serotonin metabolism in rhesus macaques . A systematic search was performed for DNA sequence variation in genes regulating neurotransmitter metabolism in rhesus macaque ( Macaca mulatta ) . These genes included dopamine and serotonin receptors and transporters , and tyrosine hydroxylase . A total of 13 single nucleotide polymorphisms in five different genes were identified , namely : P21728 ( -244T- > G ) , q = 0.45 ; P21728 ( -179C- > T ) , q = 0.19 ; P21728 ( -127G- > A ) , q = 0.25 ; P21728 ( -11T- > G ) , q = 0.08 ; P21728 ( -81C- > T ) , q = 0.19 ; P35462 ( 248G- > A ) , q= 0.08 ; P35462 ( 341G- > C ) , q = 0.11 ; P35462 ( 377A- > G ) , q = 0.19 ; P35462 ( 403C- > T ; A59V ) , q= 0.11 ; P21917 ( 2608G- > A ) , q= 0.48 ; P28221 ( -506G- > T ) , q = 0.47 ; P28221 ( -173C- > T ) , q = 0.47 ; and HTT ( 340G- > A ) , q = 0.39 . The nucleotide positions listed correspond to the human homologs . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . P80511 suppresses pro-inflammatory , but not pro-thrombotic functions of serum amyloid A . P80511 is elevated in the circulation in patients with chronic inflammatory diseases and recent studies indicate pleiotropic functions . Serum amyloid A induces monocyte cytokines and tissue factor . P80511 did not stimulate P05231 , P10145 , IL-1β or P01375 -α production by human peripheral blood mononuclear cells but low amounts consistently reduced cytokine mRNA and protein levels induced by serum amyloid A , by ∼49 % and ∼46 % , respectively . However , P80511 did not affect serum amyloid A-induced monocyte tissue factor . In marked contrast , LPS-induced cytokines or tissue factor were not suppressed by P80511 . P80511 did not alter cytokine mRNA stability or the cytokine secretory pathway . P80511 and serum amyloid A did not appear to form complexes and although they may have common receptors , suppression was unlikely via receptor competition . Serum amyloid A induces cytokines via activation of NF-κB and the MAPK pathways . P80511 reduced serum amyloid A- , but not LPS-induced P27361 /2 phosphorylation to baseline . It did not affect JNK or p38 phosphorylation or the NF-κB pathway . Reduction in P27361 /2 phosphorylation by P80511 was unlikely due to changes in intracellular reactive oxygen species , Ca(2+) flux or to recruitment of phosphatases . We suggest that P80511 may modulate sterile inflammation by blunting pro-inflammatory properties of lipid-poor serum amyloid A deposited in chronic lesions where both proteins are elevated as a consequence of macrophage activation . Serotonin- and two putative serotonin receptors-like immunohistochemical reactivities in the ground crickets Dianemobius nigrofasciatus and Allonemobius allardi . Serotonin ( 5-hydroxytryptamine ; 5-HT ) - and two putative serotonin receptors , P08908 - and P28222 -like , immunohistochemical reactivities were investigated in the cephalic ganglia of two ground crickets , Dianemobius nigrofasciatus and Allonemobius allardi . 5-HT-ir was strongly expressed in the central body , accessory medulla region of the optic lobe , frontal ganglion , posterior cortex of the protocerebrum , dorsolateral region of the protocerebrum , and the suboesphageal ganglion ( SOG ) in both crickets . However , P08908 -ir and P28222 -ir showed quite mutually distinct patterns that were also distinct from 5-HT-ir . P08908 -ir was located in the pars intercerebralis , dorsolateral region of the protocerebrum , optic tract , optic lobe , and the midline of the SOG in both crickets . P28222 -ir was located in the pars intercerebralis and dorsolateral region of the protocerebrum , and detected weakly in the optic lobe , tritocerebrum , and the midline of the SOG in both crickets . Interspecific differences were observed with P08908 -ir . P08908 -ir was expressed weakly in two neurons in the mandibular neuromere of the SOG in D. nigrofasciatus , while it was expressed strongly in the tritocerebrum , mandibular neuromere , and maxillary neuromere of the SOG in A. allardi and co-localized with O15516 -ir ( P49759 -ir ) . 5HT-1B-ir was co-localized with P49759 -ir in the tritocerebrum , mandibular neuromere , and maxillary neuromere of the SOG when double-labeling was conducted in both crickets . These results indicated that 5-HT and both types of 5-HT receptors may regulate circadian photo-entrainment or photoperiodism in A. allardi , while only P28222 may be involved in circadian photo-entrainment or photoperiodism in D. nigrofasciatus . P01258 gene-related peptide inhibits human immunodeficiency type 1 transmission by Langerhans cells via an autocrine/paracrine feedback mechanism . AIM : Peripheral neurones innervating mucosal epithelia are in direct contact with resident immune cells , including Langerhans cells ( LCs ) . Such neurones secrete the neuropeptide calcitonin gene-related peptide ( P80511 ) that modulates LCs function . We recently found that P80511 strongly inhibits human immunodeficiency virus type 1 ( HIV-1 ) transmission , by interfering with multiple steps of mucosal LC-mediated HIV-1 transfer , including increased expression of the LC-specific lectin langerin . Herein , we investigated the anti-HIV-1 mechanism of P80511 . METHODS : In the presence of P80511 , HIV-1 transfer from LCs to P01730 + T cells was tested with viral clones using either the HIV-1 co-receptor P51681 ( R5 ) or P61073 ( X4 ) . Surface expression of P51681 , P61073 and langerin was evaluated by flow cytometry . P80511 secretion by LCs was measured with an enzyme immunoassay . Expression of the multimeric P80511 receptor was examined by quantitative real-time RT-PCR and immuno-fluorescent microscopy . RESULTS : P01258 gene-related peptide decreased transfer of HIV-1 R5 , but increased that of X4 . These opposing effects correlated with decreased P51681 vs. increased P61073 surface expression in LCs . Inhibition of HIV-1 R5 transfer by P80511 involved signal transducer and activator of transcription 4 ( Q14765 ) activation . Both α P80511 and β P80511 were similarly efficient in decreasing HIV-1 R5 transfer and increasing langerin expression . LCs secreted low basal levels of endogenous P80511 , which increased markedly following P80511 treatment . P80511 also increased expression of its cognate receptor in LCs . CONCLUSION : P80511 engages a positive feedback mechanism that would further enhance its anti-HIV-1 activity . This information might be relevant for the therapeutic use of P80511 as a prophylactic agent against HIV-1 . Sequence and functional analysis of cloned guinea pig and rat serotonin P28221 receptors : common pharmacological features within the P28221 receptor subfamily . This study was undertaken to investigate the pharmacology of cloned guinea pig and rat 5-hydroxytryptamine ( serotonin ; 5-HT ) 1D receptor sites . Guinea pig , rat , and mouse P28221 receptor genes were cloned , and their amino acid sequences were compared with those of the human , dog , and rabbit . The overall amino acid sequence identity between these P28221 receptors is high and varies between 86 and 99 % . The sequence homology is slightly more divergent ( 13-27 % ) in the N-terminal extracellular region of these P28221 receptors . Guinea pig and rat P28221 receptors , stably and separately expressed in rat P13671 glial cells , are negatively coupled to cyclic AMP formation upon stimulation with agonists , as previously found for cloned human P28221 receptor sites . The cyclic AMP data show some common pharmacological features for the P28221 receptors of guinea pig , rat , and human : an almost similar rank order of potency for the investigated P28221 receptor agonists , stereoselectivity for the binding affinity and agonist potency of R(+)-8-hydroxy-2-(di-n-propylamino)tetralin , and equal P28221 receptor-mediated antagonist potency for methiothepin and the 5-HT2 receptor antagonists ritanserin and ketanserin . In conclusion , the pharmacology of the cloned P28221 receptor subtype seems , unlike the P28222 receptor subtype , conserved among various mammal species such as the human , guinea pig , and rat . Ultraviolet B irradiation reduces the expression of adiponectin in ovarial adipose tissues through endocrine actions of calcitonin gene-related peptide-induced serum amyloid A . Ultraviolet ( UV ) B irradiation decreases blood adiponectin levels , but the mechanism is not well understood . This study investigated how UVB irradiation reduces adiponectin expression in ovarial adipose tissues . Female Hos:HR-1 hairless mice were exposed to UVB ( 1.6 J/cm(2) ) irradiation and were killed 24 h later . UVB irradiation decreased the adiponectin protein level in the serum and the adiponectin mRNA level in ovarial adipose tissues . UVB irradiation also decreased the mRNA levels of peroxisome proliferator-activated receptor ( Q07869 ) γ , CCAAT/enhancer binding protein ( C/EBP ) α , C/EBPβ , and fatty acid binding protein 4 ( aP2 ) in ovarial adipose tissues . In contrast , UVB irradiation increased the mRNA levels of interleukin ( IL ) -6 and monocyte chemoattractant protein ( MCP ) -1 in ovarial adipose tissues . In the serum and liver , the levels of serum amyloid A ( P0DJI8 ) , involved in PPARγ , C/EBPα , C/EBPβ , aP2 , P05231 , and P13500 regulation , increased after UVB irradiation . The P0DJI8 gene is regulated by IL-1β , P05231 , and tumor necrosis factor-α , but only P05231 expression increased in the liver after UVB irradiation . Additionally , in the liver , hypothalamus , and epidermis , UVB irradiation increased the expression of calcitonin gene-related peptide ( P80511 ) , which upregulates P0DJI8 in the liver . Collectively , our results suggest that the P80511 signal induced by skin exposure to UVB transfers to the liver , possibly through the brain , and increases P0DJI8 production via P05231 in the liver . In turn , serum P0DJI8 acts in an endocrine manner to decreases the serum adiponectin level by downregulating factors that regulate adiponectin expression in adipose tissues .

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interacts_with DB00328?

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DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . Clathrin-dependent internalization of the angiotensin II AT₁A receptor links receptor internalization to P35354 protein expression in rat aortic vascular smooth muscle cells . The major effects of Angiotensin II ( AngII ) in vascular tissue are mediated by AngII AT1A receptor activation . Certain effects initiated by AT1A receptor activation require receptor internalization . In rat aortic vascular smooth muscle cells ( RASMC ) , AngII stimulates cyclooxygenase 2 protein expression . We have previously shown this is mediated by β-arrestin-dependent receptor internalization and NF-κB activation . In this study , a specific inhibitor of clathrin-mediated endocytosis ( CME ) , pitstop-2 , was used to test the hypothesis that clathrin-dependent internalization of activated AT1A receptor mediates NF-κB activation and subsequent cyclooxygenase 2 expression . Radioligand binding assays , real time qt-PCR and immunoblotting were used to document the effects of pitstop-2 on AngII binding and signaling in RASMC . Laser scanning confocal microscopy ( LSCM ) was used to image pitstop-2׳s effects on AT1 receptor/GFP internalization in P29320 -293 cells and p65 NF-κB nuclear localization in RASMC . Pitstop-2 significantly inhibited internalization of AT1A receptor ( 44.7 % ± 3.1 % Control vs. 13.2 % ± 8.3 % Pitstop-2 ; n=3 ) as determined by radioligand binding studies in RASMC . Studies utilizing AT1A receptor/GFP expressed in P29320 293 cells and LSCM confirmed these findings . Pitstop-2 significantly inhibited AngII-induced p65 NF-κB phosphorylation and nuclear localization , P35354 message and protein expression in RASMC without altering activation of Q8NFH3 /44 P29323 or TNFα signaling . Pitstop-2 , a specific inhibitor of clathrin-mediated endocytosis , confirms that internalization of activated AT1A receptor mediates AngII activation of cyclooxygenase 2 expression in RASMC . These data provide support for additional intracellular signaling pathways activated through β-arrestin mediated internalization of G protein-coupled receptors , such as AT1A receptors . Evidence of in vitro differential secretion of 72 and 92 kDa type IV collagenases after selective exposure to lipopolysaccharide in human fetal membranes . Premature rupture of chorioamniotic membranes complicated with intrauterine infection has been associated to degradation of extracellular matrix ( Q13201 ) , which could explain local morphological changes . We used a culture system in which the chorioamniotic membranes form two independent chambers , allowing for the selective stimulation of either the amnion ( Q9BXJ7 ) and/or the choriodecidua ( Q8NE62 ) regions . Lipopolysaccharide ( 500 ng/ml ) was added to the Q9BXJ7 and/or the Q8NE62 ; secretions and gelatinolytic activity of matrix metalloproteinase ( MMP ) -2 and P14780 were measured in both compartments by enzyme-linked immunosorbent assay ( ELISA ) and zymography . Secretions of P01033 , P16035 and Q99727 were measured by ELISA . Both metalloproteinases were immunolocalized in tissue sections . All stimulation modalities induced a similar proMMP-2 and proMMP-9 secretion pattern in the Q8NE62 with concentrations of 2.49 ng/ml and 90.91 pg/ml , respectively ; the Q9BXJ7 showed no significant changes . The active forms of both enzymes did not change with any stimulation modality . P01033 , P16035 and Q99727 secretions remained without significant changes ( P = 0.41 ) . Q13201 degradation and structural disarrangement were evident after stimulation . Secretion of proMMP-2 and proMMP-9 mainly in the Q8NE62 , presence of active forms associated to the tissue and minor changes in TIMPs secretion could favor Q13201 degradation and explain the weakening and thinning associated with the pathological rupture of chorioamniotic membranes . The protective effect of rebamipide on paracellular permeability of rat gastric epithelial cells . BACKGROUND : Barrier function in gastric epithelial cells is essential for the gastric defence mechanism against acid back-diffusion into the mucosal layer . Our previous study indicated that trans-epithelial resistance ( Q9NZ01 ) of rat gastric epithelial cells was rapidly increased when the cells were exposed to acid . This response to acid was diminished by indometacin . AIM : Evaluate the effects of a mucoprotective agent , rebamipide , on the nonsteroidal anti-inflammatory drug ( NSAID ) -induced increase of gastric epithelial permeability . METHODS : Rat gastric epithelial cells were plated on tissue culture inserts . Cells were exposed to a NSAID ( indometacin , 10-7 M ) . Trans-epithelial permeability was measured by Q9NZ01 and diffusion rate of 14C-mannitol . The effect of rebamipide was evaluated by measuring Q9NZ01 . Endogenous prostaglandin E2 ( DB00917 ) production in culture medium was also measured . RESULTS : DB00328 gradually and significantly decreased Q9NZ01 and increased 14C-manitol permeability . Rebamipide reversed the indometacin-induced changes in epithelial permeability and induced DB00917 synthesis . This induction was blocked by either indometacin or a Cyclooxygenase ( P36551 ) -2 specific inhibitor . CONCLUSIONS : P36551 inhibitors such as indometacin inhibit regulation of epithelial permeability by reducing DB00917 . P23219 has an important role in the gastric defense mechanism . Rebamipide suppressed an indometacin-induced increase in gastric epithelial permeability by increasing DB00917 levels in a P35354 dependent manner . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . LY294002 inhibits glucocorticoid-induced P35354 gene expression in cardiomyocytes through a phosphatidylinositol 3 kinase-independent mechanism . Glucocorticoids induce P35354 expression in rat cardiomyocytes . While investigating whether phosphatidylinositol 3 kinase ( PI3K ) plays a role in corticosterone ( CT ) -induced P35354 , we found that LY294002 ( LY29 ) but not wortmannin ( WM ) attenuates CT from inducing P35354 gene expression . Expression of a dominant-negative mutant of p85 subunit of PI3K failed to inhibit CT from inducing P35354 expression . CT did not activate PI3K/AKT signaling pathway whereas LY29 and WM decreased the activity of PI3K . LY303511 ( LY30 ) , a structural analogue and a negative control for PI3K inhibitory activity of LY29 , also suppressed P35354 induction . These data suggest PI3K-independent mechanisms in regulating CT-induced P35354 expression . LY29 and LY30 do not inhibit glucocorticoid receptor transactivity . Both compounds have been reported to inhibit Casein Kinase 2 activity and modulate potassium and calcium levels independent of PI3K , while LY29 has been reported to inhibit mammalian Target of DB00877 ( P42345 ) , and DNA-dependent Protein Kinase ( DNA-PK ) . Inhibitor of Casein Kinase 2 ( CK2 ) , P42345 or DNA-PK failed to prevent CT from inducing P35354 expression . DB08837 ( DB08837 ) , a potassium channel blocker , and nimodipine , a calcium channel blocker , both attenuated CT from inducing P35354 gene expression . CT was found to increase intracellular Ca(2+) concentration , which can be inhibited by LY29 , DB08837 or nimodipine . These data suggest a possible role of calcium instead of PI3K in CT-induced P35354 expression in cardiomyocytes . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Role of the JAK- P35610 pathway in protection against myocardial ischemia/reperfusion injury . The Janus kinase ( JAK ) -signal transducers and activators of transcription ( P35610 ) pathway is a stress-responsive mechanism that transduces signals from the cell surface to the nucleus , thereby modulating gene expression . Recent studies have demonstrated that myocardial ischemia and reperfusion induce rapid activation of this pathway . Although the functional consequences of this event remain to be elucidated , there is emerging evidence that JAK- P35610 signaling plays an important role in the development of the cardioprotected phenotype associated with ischemic preconditioning . Specifically , brief episodes of myocardial ischemia/reperfusion activate P23458 and O60674 , followed by recruitment of P42224 and P40763 , resulting in transcriptional upregulation of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) , which then mediate the infarct-sparing effects of the late phase of preconditioning . The present review focuses on this novel cardioprotective role of JAK- P35610 signaling and on its potential exploitation for developing therapeutic strategies aimed at limiting ischemia/reperfusion injury . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . DB08877 for the treatment of primary myelofibrosis . PURPOSE : The pharmacology , pharmacokinetics , pharmacogenomics , clinical efficacy , and safety profile of ruxolitinib for the treatment of primary myelofibrosis are reviewed . SUMMARY : DB08877 , an oral tyrosine kinase inhibitor that targets the Janus-associated kinases ( JAKs ) 1 and 2 , has been recently approved for the treatment of patients with intermediate- or high-risk myelofibrosis . Unlike previous treatment options for patients with myelofibrosis , ruxolitinib offers a targeted therapy option for these patients who often suffer with severe and debilitating symptoms associated with the disease process . After oral administration , ruxolitinib is rapidly absorbed and can be given without regard to meals . DB08877 is primarily metabolized by the cytochrome P-450 ( CYP ) 3A4 isoenzyme system ; therefore , if concomitant use with a strong P08684 inhibitor is unavoidable , an initial dosage reduction is warranted . Two Phase III randomized trials comparing ruxolitinib to either placebo or best available therapy found a rapid and sustained response in the reduction of spleen size and improvements in constitutional symptoms and quality of life , with one study demonstrating an improvement in overall survival . The most commonly reported serious adverse effects of ruxolitinib are anemia and thrombocytopenia . DB08877 is administered as an oral tablet given twice daily , with the initial starting dosage based on the baseline platelet count . Dosage reductions are based on the development of thrombocytopenia . CONCLUSION : By directly targeting both P23458 and O60674 through small-molecule inhibition , ruxolitinib elicits a reduction in splenomegaly and disease-related symptoms in patients with intermediate- or high-risk myelofibrosis while maintaining an acceptable toxicity profile and a low treatment-discontinuation rate . Functional role of wogonin in anti-angiogenesis . Constitutive activation of the Janus kinase ( JAK ) /signal transducer and activator of transcription ( P35610 ) pathway occurs commonly in cancer cells and endothelial cells , and contributes to angiogenesis . Wogonin is a compound with many biologically relevant properties . We previously reported that wogonin blocked P05231 -induced angiogenesis through suppression of P15692 expression , an important regulator of angiogenesis . However , the pathway involved in the suppressive effect of wogonin on P05231 -induced P15692 has not been completely clarified . This study aimed to investigate the molecular mechanisms participating in the suppression of wogonin on P05231 -induced P15692 in vitro , focusing on IL-6R/ P23458 / P40763 / P15692 pathway . Both P40763 siRNA and wogonin treatment resulted in an abolition of the expression of P15692 . Moreover , our data revealed that wogonin treatment after P40763 knock-down did not further suppress P15692 expression . The addition of IL-6R siRNA or wogonin resulted in a decrease in the expression level of the phosphorylated P23458 protein . Furthermore , wogonin significantly decreased the amount of phosphorylated P40763 . Finally , by EMSA , wogonin suppressed P05231 -induced P40763 binding activity in a concentration-dependent manner . Taken together , our results show that wogonin suppresses P05231 -induced P15692 by modulating the IL-6R/ P23458 / P40763 signaling pathway . Based on this study , we suggest that wogonin may provide a new potential therapeutic option for treatment of P05231 -related pathological angiogenesis . Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 and an inducible form termed P35354 . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 and P35354 . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 with the P23219 inhibitor and an R0 of 21 A for P35354 with the P35354 inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 and 18 A in P35354 . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange . IL-1beta induces P15692 , independently of DB00917 induction , mainly through the P19957 -K/ P42345 pathway in renal mesangial cells . Vascular endothelial growth factor ( P15692 ) could play a relevant role in angiogenesis associated with chronic allograft nephropathy . Interleukin-1beta ( IL-1beta ) has a key role in inflammatory response . It induces prostaglandin ( PG ) E2 , which is involved in P15692 release by some normal and tumor cells . In the present work , we studied the effect of IL-1beta on P15692 release by rat mesangial cells , the transduction signal , and whether or not DB00917 is involved in this effect . IL-1beta induced a time-dependent formation of P15692 ( analyzed by enzyme-linked immunosorbent assay ) and DB00917 ( analyzed by enzyme immunoassay ) . The latter correlated with microsomal-PGE-synthase ( mPGES ) -1 expression rather than with cyclooxygenase ( P36551 ) -2 in terms of protein , determined by Western blotting . No effect of IL-1beta on P23219 , cytosolic O14684 , or Q9H7Z7 expression was observed . Indomethacin exerted a nonsignificant effect on IL-1beta-induced P15692 , and exogenously added DB00917 exhibited a nonsignificant stimulatory effect on P15692 formation . SB 203580 , a p38 mitogen-activated protein kinase inhibitor , weakly inhibited the induction of P15692 by IL-1beta in a concentration-dependent manner , whereas LY 294002 , a phosphoinoside 3-kinase ( P19957 -K ) inhibitor , and rapamycin , a mammalian target of rapamycin ( P42345 ) inhibitor , strongly inhibited both IL-1beta- and tumor necrosis factor-alpha-induced P15692 formation in a concentration-dependent manner . DB00877 also decreased glomerular P15692 levels in the anti-Thy1.1 model of experimental glomerulonephritis . In conclusion , the P19957 -K- P42345 pathway seems to be essential in cytokine-induced release of P15692 in mesangial cells . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Dietary phytochemicals alter epigenetic events and signaling pathways for inhibition of metastasis cascade : phytoblockers of metastasis cascade . Cancer metastasis is a multistep process in which a cancer cell spreads from the site of the primary lesion , passes through the circulatory system , and establishes a secondary tumor at a new nonadjacent organ or part . Inhibition of cancer progression by dietary phytochemicals ( DPs ) offers significant promise for reducing the incidence and mortality of cancer . Consumption of DPs in the diet has been linked to a decrease in the rate of metastatic cancer in a number of preclinical animal models and human epidemiological studies . DPs have been reported to modulate the numerous biological events including epigenetic events ( noncoding micro-RNAs , histone modification , and DNA methylation ) and multiple signaling transduction pathways ( Wnt/β-catenin , Notch , Sonic hedgehog , P35354 , P00533 , MAPK- P29323 , JAK- P35610 , Akt/PI3K/ P42345 , NF-κB , AP-1 , etc. ) , which can play a key role in regulation of metastasis cascade . Extensive studies have also been performed to determine the molecular mechanisms underlying antimetastatic activity of DPs , with results indicating that these DPs have significant inhibitory activity at nearly every step of the metastatic cascade . DPs have anticancer effects by inducing apoptosis and by inhibiting cell growth , migration , invasion , and angiogenesis . Growing evidence has also shown that these natural agents potentiate the efficacy of chemotherapy and radiotherapy through the regulation of multiple signaling pathways . In this review , we discuss the variety of molecular mechanisms by which DPs regulate metastatic cascade and highlight the potentials of these DPs as promising therapeutic inhibitors of cancer . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Antitumor effect and antiangiogenic potential of the P42345 inhibitor temsirolimus against malignant pleural mesothelioma . The P42345 inhibitor temsirolimus has antitumor and antiangiogenic activity against several carcinomas , yet few reports document the efficacy of temsirolimus against malignant pleural mesothelioma ( MPM ) . Therefore , we evaluated the efficacy of temsirolimus and the antiangiogenic effect of temsirolimus in the treatment of MPM . We examined the efficacy of temsirolimus alone and the efficacy of the combination of temsirolimus and cisplatin or pemetrexed against four MPM cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ( MTT ) assay . The effect of temsirolimus on the production of proangiogenic cytokines by MPM cell lines was examined by enzyme-linked immunosorbent assay ( ELISA ) . Expression of P42345 and proangiogenic cytokines in clinical specimens from MPM patients was determined by immunohistochemistry . DB06287 inhibited cell viability and suppressed cell proliferation of all MPM cell lines . Combined treatment with temsirolimus and cisplatin inhibited the viability of all MPM cell lines more effectively than temsirolimus alone . DB06287 strongly inhibited the phosphorylation of p70s6k , a downstream molecule of P42345 , in all MPM cell lines and led to an increase in the levels of cleaved caspase-3 in the H226 and Y-meso14 cells . DB06287 also inhibited the production of vascular endothelial growth factor ( P15692 ) and platelet-derived growth factor-AA ( PDGF-AA ) . Phosphorylated P42345 and high expression of P15692 and PDGF were detected in 2 and 3 , respectively , out of the 5 MPM specimens . These results suggest that temsirolimus has activity against MPM cells by inhibition of cell proliferation and angiogenesis , and may be beneficial for a subset of MPM patients with high P42345 expression . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . DB00328 ameliorates high glucose-induced proliferation and invasion via modulation of e-cadherin in pancreatic cancer cells . DB00328 , an inhibitor of cyclooxygenase-2 ( P35354 ) , has been shown to exert anticancer effects in a variety of cancers . However , the effect and mechanism of indometacin on high glucose ( HG ) -induced proliferation and invasion of pancreatic cancer ( PC ) cells remain unclear . Multiple lines of evidence suggest that a large portion of pancreatic cancer ( PC ) patients suffer from either diabetes or HG which contributing PC progression . In this study , we report that indometacin down-regulated HG-induced proliferation and invasion via up-regulating P12830 but not P35354 in PC cells . Additionally , the P12830 transcriptional repressors , Snail and Slug , were also involved in the process . Furthermore , the proliferation and invasion of PC cells , incubated in HG medium and treated with indometacin were significantly increased when P12830 was knocked down ( Si-E-cad ) . Moreover , the protein levels of P08253 , P14780 , and P15692 were increased in PC cells transfected with Si-E-cad . Finally , the activation of the PI3K/AKT/GSK-3β signaling pathway was demonstrated to be involved in indometacin reversing HG-induced cell proliferation and invasion in PC cells . In conclusion , these results suggest that indometacin plays a key role in down-regulating HG-induced proliferation and invasion in PC cells . Our findings indicate that indometacin could be used as a novel therapeutic strategy to treat PC patients who simultaneously suffer from diabetes or HG . Some studies on spontaneous Hymenolepis diminuta infection in laboratory rats . Hymenolepis diminuta is a tapeworm that occurs worldwide . It is known to be found commonly in areas where large amounts of food grains or other dry feed products , which are the favorite foods for rats . Transmission of disease to human is uncommon ; however , it may be a serious threat for population who are living in rural areas which are suffering from excessive rodents . Here , this study had done on spontaneous H. diminuta infection in laboratory rats as a model . Out of thirty five adult laboratory rats investigated for parasitic diseases only nine ( 25.71 % ) were diagnosed positive for spontaneous H. diminuta infection . Four of them ( 44.44 % ) were found losing of weight and lacking of motility , while the others were normal . On microscopic examination , H. diminuta eggs had been found in their stool . On autopsy , small intestines were found to contain from 5-6 multi-segmented tapeworms in each rat . Histopathologically , intestinal lumen showed varying sections of H. diminuta segments with serrated borders . H. diminuta infection caused multiple mucosal ulcers with absence of intestinal villi from the surface epithelium and excessive mucin . Moreover , inflammatory cells infiltration in the connective tissue core of the villi . Furthermore , the Toluidine blue stain showed that there are Mastiocytosis . Additionally , there were goblet cells hyperplasia on using DB00233 . Moreover , there were high expression of cyclooxygenase 2 ( P35354 ) , tumor necrosis factor-α ( P01375 -α ) and inducible Nitric-Oxide Synthase ( iNOs ) . This implicate , strong correlation between P35354 , P01375 -α and iNOs expression and inflammation induced by H. diminuta . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Gremlin gene expression in bovine retinal pericytes exposed to elevated glucose . AIM : To assess the influence of high extracellular glucose on the expression of the bone morphogenetic protein ( BMP ) antagonist , gremlin , in cultured bovine retinal pericytes ( BRPC ) . METHODS : BRPC were cultured under conditions of 5 mM and 30 mM d-glucose for 7 days and total RNA was isolated . Gremlin mRNA levels were correlated , by RT-PCR , with other genes implicated in the pathogenesis of diabetic retinopathy and the signalling pathways in high glucose induced gremlin expression were probed using physiological inhibitors . Gremlin expression was also examined in the retina of streptozotocin induced diabetic mice . RESULTS : High glucose stimulated a striking increase in BRPC gremlin mRNA levels in parallel with increases in mRNA for the growth factors vascular endothelial growth factor ( P15692 ) , transforming growth factor beta ( TGFbeta ) , and connective tissue growth factor ( P29279 ) and changes in other genes including fibronectin and plasminogen activator inhibitor-1 ( P05121 ) . High glucose triggered gremlin expression was modulated by anti-TGFbeta antibody , by the uncoupler of oxidative phosphorylation , CCCP , and by inhibition of Q96HU1 -kinase ( MAPK ) activation . Striking gremlin expression was observed in the outer retina of diabetic mice and also at the level of the vascular wall . CONCLUSIONS : Gremlin gene expression is induced in BRPC in response to elevated glucose and in the retina of the streptozotocin induced diabetic mouse . Its expression is modulated by hyperglycaemic induction of the MAPK , reactive oxygen species , and TGFbeta pathways , all of which are reported to have a role in diabetic fibrotic disease . This implicates a role for gremlin in the pathogenesis of diabetic retinopathy . Preclinical in vivo evaluation of rapamycin in human malignant peripheral nerve sheath explant xenograft . Neurofibromatosis type 1 ( P21359 ) patients are prone to the development of malignant tumors , the most common being Malignant Peripheral Nerve Sheath Tumor ( MPNST ) . P21359 -MPNST patients have an overall poor survival due to systemic metastasis . Currently , the management of MPNSTs includes surgery and radiation ; however , conventional chemotherapy is not very effective , underscoring the need for effective biologically-targeted therapies . Recently , the P21359 gene product , neurofibromin , was shown to negatively regulate the phosphoinositide-3-kinase ( PI3K ) /Protein Kinase-B ( Akt ) /mammalian Target Of DB00877 ( P42345 ) pathway , with loss of neurofibromin expression in established human MPNST cell lines associated with high levels of P42345 activity . We developed and characterized a human P21359 -MPNST explant grown subcutaneously in NOD-SCID mice , to evaluate the effect of the P42345 inhibitor rapamycin . We demonstrate that rapamycin significantly inhibited human P21359 -MPNST P42345 pathway activation and explant growth in vivo at doses as low as 1.0 mg/kg/day , without systemic toxicities . While rapamycin was effective at reducing P21359 -MPNST proliferation and angiogenesis , with decreased CyclinD1 and P15692 respectively , there was no increase in tumor apoptosis . DB00877 effectively decreased activation of S6 downstream of P42345 , but there was accompanied increased Akt activation . This study demonstrates the therapeutic potential and limitations of rapamycin in P21359 -associated , and likely sporadic , MPNSTs . Evaluation of pharmacological profile of meloxicam as an anti-inflammatory agent , with particular reference to its relative selectivity for cyclooxygenase-2 over cyclooxygenase-1 . We studied the anti-inflammatory activity of meloxicam on rat carrageenin-induced pleurisy and its toxicity for rat gastric mucosa , relative to its in vitro inhibitory potency against partially purified cyclooxygenase ( P36551 ) -1 and P35354 preparations in order to clarify the pharmacological profile of the compound as an anti-inflammatory agent . In rat carrageenin-induced pleurisy , the plasma exudation rate peaked at 5 h , at which time P35354 was detectable in cells from the pleural exudate . Meloxicam and piroxicam ( 1 and 3 mg/kg ) and NS-398 ( 3 mg/kg ) showed almost equal anti-inflammatory potency against 5-hour pleurisy . A single oral administration of the compounds caused a dose-dependent increase in the number of rats with gastric mucosal erosion . The ED50 value for meloxicam ( 5.92 mg/kg ) was significantly higher than that for piroxicam ( 1.76 mg/kg ) , indicating that meloxicam is safer . DB00328 showed intermediate safety ( 2.59 mg/kg ) . In in vitro experiments , indometacin inhibited P23219 about 1.7 times more potently than P35354 . NS-398 inhibited P35354 with an IC50 of 0.32 microM , but never affected P23219 activity , even at 100 microM . In the same assay system , meloxicam inhibited P35354 about 12 times more selectively than P23219 . Piroxicam , however , inhibited both isoforms almost equally . These results indicate that meloxicam is a potent anti-inflammatory agent with low gastric toxicity . One reason for its in vivo pharmacological profile may be related to its relative selectivity for P35354 over P23219 . Thus , meloxicam may belong to a group of P35354 selective anti-inflammatory agents with a better safety profile than conventional P23219 and P35354 nonselective anti-inflammatory agents .

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Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Identification and analysis of P22680 , P05093 , Q6UW02 , Q02318 and Q16850 in cynomolgus macaques . Cytochromes P450 ( P450 ) are important for not only drug metabolism and toxicity , but also biosynthesis and metabolism of cholesterol and bile acids , and steroid synthesis . In cynomolgus macaques , widely used in biomedical research , we have characterized P450 cDNAs , which were isolated as expressed sequence tags of cynomolgus macaque liver . In this study , cynomolgus P22680 , P05093 , Q6UW02 , Q02318 and Q16850 cDNAs were characterized by sequence analysis , phylogenetic analysis and tissue expression pattern . By sequence analysis , these five cynomolgus P450s had high sequence identities ( 94-99 % ) to the human orthologs in amino acids . By phylogenetic analysis , each cynomolgus P450 was more closely related to the human ortholog as compared with the dog or rat ortholog . By quantitative polymerase chain reaction , among the 10 tissue types , P22680 and P05093 mRNAs were preferentially expressed in liver and adrenal gland , respectively . Cynomolgus Q02318 and Q16850 mRNAs were most abundantly expressed in liver and testis , respectively . Cynomolgus Q6UW02 mRNA was expressed in all the tissues , including brain and liver . Tissue expression patterns of each cynomolgus P450 were generally similar to that of the human ortholog . These results suggest the molecular similarities of P22680 , P05093 , Q6UW02 , Q02318 and Q16850 between cynomolgus macaques and humans . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . Q9UGN5 interacts with Q15669 -1 and regulates expression of surfactant protein-B . P43699 ( Q15669 -1/Nkx-2.1 ) plays a critical role in lung morphogenesis and regulates the expression of lung-specific genes , including the surfactant proteins required for pulmonary function after birth . The activity of Q15669 -1 is influenced by its interactions with other transcription factors and coactivators , including CBP/p300 and Q15788 . In this study , we have identified poly(ADP-ribose) polymerases ( Q9UGN5 and P09874 ) as Q15669 -1 interacting proteins that influence its transcriptional activity . Endogenous Q9UGN5 was coimmunoprecipitated from transformed mouse lung epithelial cell ( MLE15 ) extracts with Q15669 -1 and was identified by mass spectrometry . P09874 and P12956 / P13010 were also coimmunoprecipitated from the cell extracts with Q15669 -1 . The E domain of Q9UGN5 interacted via the C-terminal domain of Q15669 -1 . Both P09874 and Q9UGN5 enhanced the activity of the promoter of surfactant protein-B ( Sftpb gene ) but not other surfactant proteins in vitro . Q9UGN5 was selectively expressed in epithelial cells of the conducting and peripheral lung tubules of the fetal mouse lung from embryonic day 12.5 and was detected in bronchial epithelial cells in the adult lung at cellular sites consistent with that of surfactant protein B . Q9UGN5 and P09874 interact with Q15669 -1 and regulate the expression of surfactant protein B , a protein required for lung function . P10275 inactivation contributes to antitumor efficacy of 17{alpha}-hydroxylase/17,20-lyase inhibitor 3beta-hydroxy-17-(1H-benzimidazole-1-yl)androsta-5,16-diene in prostate cancer . We previously reported that our novel compound 3beta-hydroxy-17-(1H-benzimidazole-1-yl)androsta-5,16-diene ( VN/124-1 ) is a potent 17alpha-hydroxylase/17,20-lyase ( P05093 ) inhibitor/antiandrogen and strongly inhibits the formation and proliferation of human prostate cancer LAPC4 tumor xenografts in severe combined immunodeficient mice . In this study , we report that VN/124-1 and other novel P05093 inhibitors also cause down-regulation of androgen receptor ( AR ) protein expression in vitro and in vivo . This mechanism of action seems to contribute to their antitumor efficacy . We compared the in vivo antitumor efficacy of VN/124-1 with that of castration and a clinically used antiandrogen , DB01128 , and show that VN/124-1 is more potent than castration in the LAPC4 xenograft model . Treatment with VN/124-1 ( 0.13 mmol/kg twice daily ) was also very effective in preventing the formation of LAPC4 tumors ( 6.94 versus 2410.28 mm(3) in control group ) . VN/124-1 ( 0.13 mmol/kg twice daily ) and VN/124-1 ( 0.13 mmol/kg twice daily ) + castration induced regression of LAPC4 tumor xenografts by 26.55 % and 60.67 % , respectively . Treatments with DB01128 ( 0.13 mmol/kg twice daily ) or castration caused significant tumor suppression compared with control . Furthermore , treatment with VN/124-1 caused marked down-regulation of AR protein expression , in contrast to treatments with DB01128 or castration that caused significant up-regulation of AR protein expression . The results suggest that VN/124-1 acts by several mechanisms ( P05093 inhibition , competitive inhibition , and down-regulation of the AR ) . These actions contribute to inhibition of the formation of LAPC4 tumors and cause regression of growth of established tumors . VN/124-1 is more efficacious than castration in the LAPC4 xenograft model , suggesting that the compound has potential for the treatment of prostate cancer . Quantification of rifampicin in human plasma and cerebrospinal fluid by a highly sensitive and rapid liquid chromatographic-tandem mass spectrometric method . A highly sensitive and rapid liquid chromatography tandem mass spectrometry ( LC-MS/MS ) method has been developed to measure the levels of the antitubercular drug rifampicin ( Q9HBH0 ) in human plasma and cerebrospinal fluid ( P04141 ) . The analyte and internal standard ( IS ) were isolated from plasma and P04141 by a simple organic solvent based precipitation of proteins followed by centrifugation . Detection was carried out by electrospray positive ionization mass spectrometry in the multiple-reaction monitoring ( MRM ) mode . The assay was linear in the concentration range 25-6400 ng/mL with intra- and inter-day precision of < 7 % and < 8 % , respectively . The validated method was applied to the study of Q9HBH0 pharmacokinetics in human P04141 and plasma over 25 h period after a 10 mg/kg oral dose . Diet induced regulation of genes involved in cholesterol metabolism in rat liver parenchymal and Kupffer cells . BACKGROUND/AIMS : Feeding rodents atherogenic diets enriched in cholesterol or cholic acid changes hepatic cholesterol metabolism . In the present study , the effect of an atherogenic diet enriched in cholesterol and cholic acid on cellular hepatic cholesterol metabolism was studied . METHODS : Gene and protein expression analysis was performed on parenchymal , endothelial , and Kupffer cells isolated from rats fed a chow or atherogenic diet using quantitative real-time PCR and immunoblotting , respectively . RESULTS : The atherogenic diet raised the serum cholesterol concentration 11-fold , mostly in the VLDL fraction , and led to heavy lipid loading of rat liver parenchymal and Kupffer cells . Only moderate changes in the expression of genes involved in cholesterol metabolism were observed in parenchymal cells on the diet , while Q07869 delta expression was 6.8-fold decreased . Kupffer cells , however , showed a highly adaptive response with a 2- to 9-fold induction of Q8WTV0 , O95477 , and Q9H222 / Q9UBA6 , and an 82-fold induction in P22680 mRNA expression , respectively . CONCLUSIONS : Heavy lipid loading of parenchymal cells leads to moderate gene expression changes , while Kupffer cells respond in a highly adaptive fashion by stimulating the expression of genes involved in cholesterol metabolism and transport . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . DB01045 Does not Significantly Affect the Expression of Small Heterodimer Partner in Primary Human Hepatocytes . The small/short heterodimer partner ( Q15466 , Q15466 ) is a nuclear receptor corepressor lacking a DNA binding domain . Q15466 is induced by bile acid-activated farnesoid X receptor ( Q96RI1 ) resulting in P22680 gene suppression . In contrast , O75469 ( O75469 ) activation by its ligands was recently suggested to inhibit Q15466 gene transactivation to maximize the induction of O75469 target genes . However , there are also conflicting reports in literature whether O75469 or rodent Pxr activation down-regulates Q15466 /Shp expression . Moreover , the O75469 -mediated regulation of the Q15466 gene has been studied only at the Q15466 mRNA and transactivation ( gene reporter assay ) levels . In this study , we studied the effect of rifampicin , a prototype O75469 ligand , on Q15466 mRNA , and protein expression in three primary human hepatocyte cultures . We found that Q15466 mRNA is not systematically down-regulated in hepatocyte in culture after 24 h treatment with rifampicin . Consistently , we did not observe down-regulation of Q15466 protein in primary human hepatocytes after 24 and 48 h of incubation with rifampicin . We can conclude that although we observed slight down-regulation of Q15466 mRNA and protein in several hepatocyte preparations , the phenomenon is unlikely critical for O75469 -mediated induction of its target genes . Molecular mechanisms involved in the growth stimulation of breast cancer cells by leptin . Obesity is a risk factor for breast cancer in postmenopausal women . Leptin , an adipocyte-derived cytokine , elicits proliferative effects in some cell types and potentially stimulates the growth of mammary epithelium . Here we show that leptin induced time- and dose-dependent signal transducer and activator of transcription 3 ( P40763 ) phosphorylation and extracellular signal-regulated kinase ( P29323 ) 1/2 kinase activation in breast carcinoma cells . Blocking P40763 phosphorylation with a specific inhibitor , AG490 , abolished leptin-induced proliferation of MCF-7 cells , whereas blocking P27361 /2 activation by a specific P27361 /2 kinase inhibitor , U0126 , did not result in any significant changes in leptin-induced cell proliferation . Our experiments also showed that one member of the P52701 family of steroid receptor coactivators , steroid receptor coactivator ( P12931 ) -1 , but not glucocorticoid receptor interacting protein 1 ( GRIP1 ) or amplified in breast cancer 1 ( Q9Y6Q9 ) , also functioned in gene transactivation in response to leptin treatment . O60760 pull-down experiments showed that Q15788 physically interacted with the activation domain of P40763 and that chromatin immunoprecipitation experiments detected the occupancy of Q15788 , but not GRIP1 or Q9Y6Q9 , on the promoter of P40763 target genes . Our experiments collectively showed that Q15788 is involved in P40763 signaling pathway that is implicated in leptin-stimulated cell growth . Down-regulation of RXRalpha expression is essential for neutrophil development from granulocyte/monocyte progenitors . Neutrophil granulocytes ( Gs ) represent highly abundant and short-lived leukocytes that are constantly regenerated from a small pool of myeloid committed progenitors . Nuclear receptor ( NR ) family members are ligand-activated transcription factors that play key roles in cellular proliferation and differentiation processes including myelopoiesis . P19793 ( RXRalpha ) represents the predominant NR types I and II homo- and heterodimerization partner in myeloid cells . Here we show that human myeloid progenitors express RXRalpha protein at sustained high levels during macrophage colony-stimulating factor ( P09603 ) -induced monopoiesis . In sharp contrast , RXRalpha is down-regulated during G- P04141 -dependent late-stage neutrophil differentiation from myeloid progenitors . Down-regulation of RXRalpha is critically required for neutrophil development since ectopic RXRalpha inhibited granulopoiesis by impairing proliferation and differentiation . Moreover , ectopic RXRalpha was sufficient to redirect G- P04141 -dependent granulocyte differentiation to the monocyte lineage and to promote P09603 -induced monopoiesis . Functional genetic interference with RXRalpha signaling in hematopoietic progenitor/stem cells using a dominant-negative RXRalpha promoted the generation of late-stage granulocytes in human cultures in vitro and in reconstituted mice in vivo . Therefore , our data suggest that RXRalpha down-regulation is a critical requirement for the generation of neutrophil granulocytes . Attenuation of anti-tuberculosis therapy induced hepatotoxicity by Spirulina fusiformis , a candidate food supplement . Therapy using Isoniazid ( DB00951 ) and DB01045 ( Q9HBH0 ) leads to induction of hepatotoxicity in some individuals undergoing anti-tuberculosis treatment . In this study , we assessed the effect of Spirulina fusiformis on DB00951 and Q9HBH0 induced hepatotoxicity in rats compared with hepatoprotective drug Silymarin . Induction of hepatotoxicity was measured by changes in the liver marker enzymes ( aspartate transaminase , alanine transaminase , and alkaline phosphatase ) . The antioxidant status was also analyzed in liver tissue homogenate and plasma by measurement of superoxide dismutase , catalase , glutathione-S-transferase , glutathione reductase , and lipid peroxidation levels . We also aimed to study the binding and interactions of the transcription factors Pregnane X Receptor ( O75469 ) and Farnesoid X Receptor ( Q96RI1 ) with DB00951 , Q9HBH0 , and representative active compounds of Spirulina fusiformis by in silico methods . The administration of DB00951 and Q9HBH0 resulted in significant ( p < 0.05 ) decrease in the antioxidant levels and total protein levels . There was also a significant ( p < 0.05 ) increase in the levels of liver marker enzymes . Spirulina fusiformis was seen to protect the parameters from significant changes upon challenge with DB00951 and Q9HBH0 in a dose-dependent manner . This was corroborated by histological examination of the liver . The results of the in silico analyses further support the wet lab results . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Novel cinnamyl hydroxyamides and 2-aminoanilides as histone deacetylase inhibitors : apoptotic induction and cytodifferentiation activity . Four novel series of cinnamyl-containing histone deacetylase ( HDAC ) inhibitors 1-4 are described , containing hydroxamate ( 1 and 3 ) or 2-aminoanilide ( 2 and 4 ) derivatives . When screened against class I ( maize HD1-B and human Q13547 ) and class II ( maize HD1-A and human P56524 ) HDACs , most hydroxamates and 2-aminoanilides displayed potent and selective inhibition toward class I enzymes . Immunoblotting analyses performed in U937 leukemia cells generally revealed high acetyl-H3 and low acetyl-α-tubulin levels . Exceptions are compounds 3 f-i , 3 m-o , and 4 k , which showed higher tubulin acetylation than DB02546 . In U937 cells , cell-cycle blockade in either the G₂/M or G₁/S phase was observed with 1-4 . Five hydroxamates ( compounds 1 h-l ) effected a two- to greater than threefold greater percent apoptosis than DB02546 , and in the CD11c cytodifferentiation test some 2-aminoanilides belonging to both series 2 and 4 were more active than MS-275 . The highest-scoring derivatives in terms of apoptosis ( 1 k , 1 l ) or cytodifferentiation ( 2 c , 4 n ) also showed antiproliferative activity in U937 cells , thus representing valuable tools for study in other cancer contexts . Effects of DB01045 , a potent inducer of drug-metabolizing enzymes and an inhibitor of Q9Y6L6 /3 transport , on the single dose pharmacokinetics of anacetrapib . Anacetrapib is a novel cholesteryl ester transfer protein ( P11597 ) inhibitor in development for treatment of dyslipidemia . This open-label , fixed-sequence , 3-period study was intended to evaluate the potential of anacetrapib to be a victim of Q9Y6L6 /3 inhibition and strong CYP3A induction using acute and chronic dosing of rifampin , respectively , as a probe . In this study , 16 healthy subjects received 100 mg anacetrapib administered without rifampin ( Day 1 , Period 1 ) , with single-dose ( SD ) 600 mg rifampin ( Day 1 , Period 2 ) , and with multiple-dose ( MD ) 600 mg rifampin for 20 days ( Day 14 , Period 3 ) . Log-transformed anacetrapib AUC0-∞ and Cmax were analyzed by a linear mixed effects model . The GMRs and 90 % CIs for anacetrapib AUC0-∞ and Cmax were 1.25 ( 1.04 , 1.51 ) and 1.43 ( 1.13 , 1.82 ) for SD rifampin ( Period 2/Period 1 ) and 0.35 ( 0.29 , 0.42 ) and 0.26 ( 0.21 , 0.32 ) for MD rifampin ( Period 3/Period 1 ) , respectively . Anacetrapib was generally well tolerated in both the absence/presence of SD and MD rifampin . In conclusion , treatment with SD rifampin , which inhibits the Q9Y6L6 /3 transporter system , did not substantially influence the SD pharmacokinetics of anacetrapib , while chronic ( 20 days ) administration of rifampin , which strongly induces CYP3A isozymes , reduced mean systemic exposure to SD anacetrapib by 65 % . Transcriptional repression of mitochondrial function in aging : a novel role for the silencing mediator of retinoid and thyroid hormone receptors co-repressor . SIGNIFICANCE : Mitochondrial function plays an important role in metabolic homeostasis and has been implicated in aging . Although there is still ongoing debate regarding whether mitochondrion-derived oxidative stress is causative to the aging process , interventions that increase oxidative metabolism and antioxidant pathways in animal models protect against age-related deterioration , such as metabolic diseases and neurodegenerative disorders . RECENT ADVANCES : One of the well-characterized transcriptional networks known to improve mitochondrial activity is mediated by transcriptional co-activator peroxisome proliferator-activated receptor gamma co-activator 1α ( P20142 -1α ) , which is activated by AMP-activated protein kinase ( AMPK ) and sirtuin 1 ( Q96EB6 ) , two of the major energy sensing molecules that are responsible for the longevity effect of caloric restriction in certain model systems . P20142 -1α co-activates several nuclear receptors , notably members of the peroxisome proliferator-activated receptor ( Q07869 ) family , which are key regulators of mitochondrial oxidative metabolism . CRITICAL ISSUES : Although the AMPK/ Q96EB6 - P20142 -1α- Q07869 axis plays a prominent role in activating mitochondrial functions , their activities are down-regulated in older animals , suggesting the involvement of dominant negative regulatory mechanisms in the process of aging . FUTURE DIRECTIONS : In this review , we will discuss the role of a transcriptional co-repressor , silencing mediator of retinoid and thyroid hormone receptors ( Q9Y618 ) , whose activity and expression are increased with age , as a negative regulator of mitochondrial function that promotes aging and age-related metabolic diseases . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . Constitutive activation of the mitogen-activated protein kinase pathway impairs vitamin D signaling in human prostate epithelial cells . We studied the effect of prolonged activation of mitogen-activated protein kinase ( MAPK ) signaling on 1,25 dihydroxyvitamin D ( 1,25(OH)(2)D(3) ) action in the immortalized human prostate epithelial cell line RWPE1 and its P01116 transformed clone RWPE2 . 1,25(OH)(2)D(3)-treatment caused growth arrest and induced gene expression in both cell lines but the response was blunted in RWPE2 cells . Vitamin D receptor ( P11473 ) levels were lower in RWPE2 cells but P11473 over-expression did not increase vitamin-D-mediated gene transcription in either cell line . In contrast , MAPK inhibition restored normal vitamin D transcriptional responses in RWPE2 cells and MAPK activation with constitutively active MEK1R4F reduced vitamin-D-regulated transcription in RWPE1 cells . 1,25(OH)(2)D(3)-mediated transcription depends upon the P11473 and its heterodimeric partner the retinoid X receptor ( RXR ) so we studied whether changes in the P11473 -RXR transcription complex occur in response to MAPK activation . Mutation of putative phosphorylation sites in the activation function 1 ( P38484 ) domain ( S32A , T82A ) of RXRalpha restored 1,25(OH)(2)D(3)-mediated transactivation in RWPE2 cells . Mammalian two-hybrid and co-immunoprecipitation assays revealed a vitamin-D-independent interaction between steroid receptor co-activator-1 ( Q15788 ) and RXRalpha that was reduced by MAPK activation and was restored in RWPE2 cells by mutating S32 and T82 in the RXRalpha P38484 domain . Our data show that a common contributor to cancer development , prolonged activation of MAPK signaling , impairs 1,25(OH)(2)D(3)-mediated transcription in prostate epithelial cells . This is due in part to the phosphorylation of critical amino acids in the RXRalpha P38484 domain and impaired co-activator recruitment . The mechanism of the G0/ P55008 cell cycle phase arrest induced by activation of O75469 in human cells . CONTEXT : O75469 ( O75469 ) is an important transcriptional regulator that plays important roles in the cell metabolism and cell growth by regulating the transcriptional of a sort of metabolizing enzymes . OBJECTIVE : To investigate whether rifampicin effected HepG2 cells growth and the inhibition was due to the G0/ P55008 phase arrest . METHODS : O75469 -knockdown experiments using RNAi showed that the cell cycle phase arrest mediated by rifampicin based on activation of O75469 . The results also indicated that cell phase arrest by rifampicin could protect cells form UVB-induced DNA damage . P19793 ( RXRα ) expression level in cells is another key factor for cell cycle phase arrest mediated by rifampicin . Both over expression and lacking expression of RXRα in cell reduced the cell arrest efficiency mediated by rifampicin . In the study , we found that rifampicin inhibited HepG2 cells growth and demonstrated that the inhibition is due to the G0/ P55008 phase arrest through flow cytometry analysis . CONCLUSION : The results showed that RXRα promote cell cycle phase transition rate of HepG2 . Competitive bind of rifampicin-activated O75469 with RXRα is one main reason to arrest cell cycle phase through inhibiting combination of RXRα with other partners . DB01045 could promote cell growth rate when RXRα expressed more excessively than O75469 in cells . A novel bile acid-activated vitamin D receptor signaling in human hepatocytes . Vitamin D receptor ( P11473 ) is activated by natural ligands , 1alpha , 25-dihydroxy-vitamin D(3) [ 1alpha,25(OH)(2)-D(3) ] and lithocholic acid ( LCA ) . Our previous study shows that P11473 is expressed in human hepatocytes , and P11473 ligands inhibit bile acid synthesis and transcription of the gene encoding cholesterol 7alpha-hydroxylase ( P22680 ) . Primary human hepatocytes were used to study LCA and 1alpha,25(OH)(2)-D(3) activation of P11473 signaling . Confocal immunofluorescent microscopy imaging and immunoblot analysis showed that LCA and 1alpha , 25(OH)(2)-D(3) induced intracellular translocation of P11473 from the cytosol to the nucleus and also plasma membrane where P11473 colocalized with caveolin-1 . P11473 ligands induced tyrosine phosphorylation of c-Src and P11473 and their interaction . Inhibition of c-Src abrogated P11473 ligand-dependent inhibition of P22680 mRNA expression . Kinase assays showed that P11473 ligands specifically activated the c-Raf/ Q02750 /2/extracellular signal-regulated kinase ( P29323 ) 1/2 pathway , which stimulates serine phosphorylation of P11473 and hepatocyte nuclear factor-4alpha , and their interaction . Mammalian two-hybrid assays showed a P11473 ligand-dependent interaction of nuclear receptor corepressor-1 and silencing mediator of retinoid and thyroid with P11473 /retinoid X receptor-alpha ( RXRalpha ) . Chromatin immunoprecipitation assays revealed that an P27361 /2 inhibitor reversed P11473 ligand-induced recruitment of P11473 , RXRalpha , and corepressors to human P22680 promoter . In conclusion , P11473 ligands activate membrane P11473 signaling to activate the Q02750 /2/ P27361 /2 pathway , which stimulates nuclear P11473 /RXRalpha recruitment of corepressors to inhibit P22680 gene transcription in human hepatocytes . This membrane P11473 -signaling pathway may be activated by bile acids to inhibit bile acid synthesis as a rapid response to protect hepatocytes from cholestatic liver injury . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Granulocyte macrophage-colony stimulating factor increases the expression of histamine and histamine receptors in monocytes/macrophages in relation to arteriosclerosis . OBJECTIVE : To study the effect of granulocyte macrophage-colony-stimulating factor ( GM- P04141 ) on histamine metabolism in arteriosclerosis , the expression of histidine decarboxylase ( HDC ; histamine-producing enzyme ) , histamine receptors 1 and 2 ( P35367 and P25021 ) , and GM- P04141 was investigated in human and mouse arteriosclerotic carotid arteries . Furthermore , the molecular mechanisms of GM- P04141 -induced HDC and P35367 expression in monocytic U937 cells were investigated . METHODS AND RESULTS : Immunohistochemistry showed that atherosclerotic human coronary and mouse ligated carotid arteries contained HDC-expressing macrophages . Gene expression of HDC , P35367 , P25021 , and GM- P04141 was also detected in the lesions . In U937 cells , GM- P04141 enhanced histamine secretion and gene expression of HDC and P35367 . A promoter assay showed that GM- P04141 enhanced gene transcription of HDC and P35367 but not P25021 . CONCLUSIONS : The present results indicate that HDC and HHR are expressed in arteriosclerotic lesion , and that GM- P04141 induces HDC and P35367 expression in monocytes . Locally produced histamine might participate in atherogenesis by affecting the expression of atherosclerosis-related genes in monocytes and smooth muscle cells . The presence of histamine-producing macrophages and gene expression of histamine receptors and GM- P04141 was demonstrated in arteriosclerotic lesions . In monocytic U937 cells , GM- P04141 upregulated the expression of histamine and P35367 . Coordinated expression of histamine and its receptors by GM- P04141 would participate in atherogenesis by affecting monocytic and SMC gene expression . LPS-induced downregulation of Q92887 and O95342 in human liver is due to a posttranscriptional process . Endotoxin-induced cholestasis in rodents is caused by hepatic downregulation of transporters , including the basolateral Na+-dependent taurocholate transporter ( ntcp ) and the canalicular bile salt export pump ( bsep ) and multidrug resistance-associated protein 2 ( mrp2 ) . Details about the regulation of the human transporter proteins during this process are lacking . We used precision-cut human and rat liver slices to study the regulation of transporter expression during LPS-induced cholestasis . We investigated the effect of LPS on nitrate/nitrite and cytokine production in relation to the expression of inducible nitric oxide synthase , Q14973 , O95342 , and Q92887 both at the level of mRNA with RT-PCR and protein using immunofluorescence microscopy . In liver slices from both species , LPS-induced expression of inducible nitric oxide synthase was detected within 1-3 h and remained increased over 24 h . In rat liver slices , this was accompanied by a significant decrease of rat ntcp and mrp2 mRNA levels , whereas bsep levels were not affected . These results are in line with previous in vivo studies and validate our liver slice technique . In LPS-treated human liver slices , Q14973 mRNA was downregulated and showed an inverse correlation with the amounts of P01375 and Il-1beta produced . In contrast , Q92887 and O95342 mRNA levels were not affected under these conditions . However , after 24-h LPS challenge , both proteins were virtually absent in human liver slices , whereas marker proteins remained detectable . In conclusion , we show that posttranscriptional mechanisms play a more prominent role in LPS-induced regulation of human Q92887 and O95342 compared with the rat transporter proteins . JTT-705 blocks cell proliferation and angiogenesis through p38 kinase/p27(kip1) and Ras/ P38936 (waf1) pathways . The excessive proliferation and migration of vascular smooth muscle cells ( SMCs ) participate in the growth and instability of atherosclerotic plaque . We examined the direct role of a newly developed chemical inhibitor of cholesteryl ester transfer protein , JTT-705 , on SMC proliferation and angiogenesis in endothelial cells ( ECs ) . JTT-705 inhibited human coronary artery SMC proliferation . JTT-705 induced the phosphorylation of p38 mitogen-activated protein kinase ( MAPK ) and extracellular-signal-regulated kinases ( P29323 ) in SMCs . In addition , the anti-proliferative effects of JTT-705 in SMCs were blocked by p38 MAPK inhibitor . JTT-705 induced the upregulation of p- P38936 (waf1) , and this effect was blocked by dominant-negative Ras ( N17 ) , but not by inhibitors of p38 MAPK or P29323 . In addition , JTT-705 also induced the upregulation of p27(kip1) , and this effect was blocked by p38 MAPK inhibitor . Interestingly , culture medium from JTT-705-treated SMCs blocked human coronary artery EC tube formation in an in vitro model of angiogenesis indirectly via a decrease in vascular endothelial growth factor ( P15692 ) from SMCs and directly via an anti-proliferative effect in ECs . JTT-705 blocked the proliferation of SMCs through the activation of p38 kinase/p27(kip1) and Ras/ P38936 (waf1) pathways , and simultaneously blocked EC tube formation associated with a decrease in P15692 production from SMCs and an anti-proliferative effect in ECs . Our results indicate that JTT-705 may induce a direct anti-atherogenic effect in addition to its inhibitory effect of P11597 activity . O75469 induces Q02318 and regulates cholesterol metabolism in the intestine . Mitochondrial sterol 27-hydroxylase ( Q02318 ) catalyzes oxidative cleavage of the sterol side chain in the bile acid biosynthetic pathway in the liver and 27-hydroxylation of cholesterol in most tissues . Recent studies suggest that 27-hydroxycholesterol ( 27-HOC ) activates liver orphan receptor alpha ( LXRalpha ) and induces the cholesterol efflux transporters O95477 and P45844 in macrophages . The steroid- and bile acid-activated pregnane X receptor ( O75469 ) plays critical roles in the detoxification of bile acids , cholesterol metabolites , and xenobiotics . The role of Q02318 in the intestine is not known . This study investigated O75469 and Q02318 regulation of cholesterol metabolism in the human intestinal cell lines Caco2 and Ls174T . A human O75469 ligand , rifampicin , induced Q02318 mRNA expression in intestine cells but not in liver cells . DB01045 induced Q02318 gene transcription , increased intracellular 27-HOC levels , and induced O95477 and P45844 mRNA expression only in intestine cells . A functional O75469 binding site was identified in the human Q02318 gene . Chromatin immunoprecipitation assays revealed that rifampicin induced the O75469 recruitment of steroid receptor coactivator 1 to Q02318 chromatin . DB04540 loading markedly increased intracellular 27-HOC levels in intestine cells . DB01045 , 27-HOC , and a potent LXRalpha agonist , T0901317 , induced O95477 and P45844 protein expression and stimulated cholesterol efflux from intestine cells to apolipoprotein A-I and HDL . This study suggests an intestine-specific O75469 / Q02318 /LXRalpha pathway that regulates intestine cholesterol efflux and HDL assembly . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . Down-regulation of organic anion transporter expression in human hepatocytes exposed to the proinflammatory cytokine interleukin 1beta . Interleukin ( IL ) 1beta is a proinflammatory cytokine known to markedly alter expression of major organic anion transporters in rodent hepatocytes . However , its effects toward human hepatic transporters remain poorly characterized . Therefore , the present study was aimed at determining IL-1beta effects on expression of organic anion transporters in primary human hepatocytes and highly differentiated human hepatoma HepaRG cells . Exposure to 1 ng/ml IL-1beta was first shown to markedly repress mRNA expression of sodium-taurocholate cotransporting polypeptide ( Q14973 ) , a major sinusoidal transporter handling bile acids , in both human hepatocytes and HepaRG cells . It concomitantly reduced Q14973 protein levels and Q14973 -mediated cellular uptake of taurocholate in HepaRG cells . Other transporters such as the influx transporters organic anion transporting polypeptide ( P46721 ) -B , Q9Y6L6 , and Q9NPD5 and the efflux pumps multidrug resistance-associated protein ( MRP ) 2 , O15438 , O15439 , and breast cancer resistance protein were also down-regulated at mRNA levels in human hepatocytes treated by IL-1beta for 24 h , and most of these transporters were similarly repressed in IL-1beta-exposed HepaRG cells ; the cytokine also reduced bile salt export pump ( O95342 ) and Q9Y6L6 protein expression in human hepatocytes . IL-1beta was further shown to activate the extracellular signal-regulated protein kinase ( P29323 ) in human hepatocytes and HepaRG cells ; however , chemical inhibition of this kinase failed to counteract repressing effects of IL-1beta toward Q14973 , O95342 , O94956 , and Q9Y6L6 . Taken together , these data indicate that IL-1beta treatment reduced expression of major organic anion transporters in human hepatic cells in an P29323 -independent manner . Such IL-1beta effects may likely participate in both cholestasis and alterations of hepatic detoxification pathways caused by inflammation in humans . Effect of active smoking on the human bronchial epithelium transcriptome . BACKGROUND : Lung cancer is the most common cause of cancer-related deaths . Tobacco smoke exposure is the strongest aetiological factor associated with lung cancer . In this study , using serial analysis of gene expression ( Q9NXZ1 ) , we comprehensively examined the effect of active smoking by comparing the transcriptomes of clinical specimens obtained from current , former and never smokers , and identified genes showing both reversible and irreversible expression changes upon smoking cessation . RESULTS : Twenty-four Q9NXZ1 profiles of the bronchial epithelium of eight current , twelve former and four never smokers were generated and analyzed . In total , 3,111,471 Q9NXZ1 tags representing over 110 thousand potentially unique transcripts were generated , comprising the largest human Q9NXZ1 study to date . We identified 1,733 constitutively expressed genes in current , former and never smoker transcriptomes . We have also identified both reversible and irreversible gene expression changes upon cessation of smoking ; reversible changes were frequently associated with either xenobiotic metabolism , nucleotide metabolism or mucus secretion . Increased expression of Q07654 , O75952 , and Q5MY95 were found to be reversible upon smoking cessation . Expression of P49841 , which regulates P35354 expression , was irreversibly decreased . P98088 expression was only partially reversed . Validation of select genes was performed using quantitative RT-PCR on a secondary cohort of nine current smokers , seven former smokers and six never smokers . CONCLUSION : Expression levels of some of the genes related to tobacco smoking return to levels similar to never smokers upon cessation of smoking , while expression of others appears to be permanently altered despite prolonged smoking cessation . These irreversible changes may account for the persistent lung cancer risk despite smoking cessation . Effect of prototypical inducing agents on P-glycoprotein and CYP3A expression in mouse tissues . P-glycoprotein ( P-gp ) and CYP3A have considerable overlap in inducers in vitro . Characterizing P-gp induction in vivo and potential coregulation with CYP3A are important goals for predicting drug interactions . This study examined P-gp expression in mouse tissues and potential coinduction with CYP3A following oral treatment with 1 of 7 prototypical inducing agents for 5 days . P-gp expression in brain or liver was not induced by any treatment as determined by Western blot , whereas dexamethasone , pregnenolone-16alpha-carbonitrile ( Q15149 ) , St . John 's wort ( SJW ) , and rifampin induced hepatic CYP3A expression . In intestine , rifampin and SJW induced P-gp expression 3.7- and 1.6-fold and CYP3A 3.5- and 2.4-fold , respectively , whereas dexamethasone and Q15149 induced CYP3A only . These observations suggest that P-gp in mouse small intestine is inducible by some , but not all , CYP3A inducers , whereas P-gp expression in liver or brain is not readily induced . Intriguingly , rifampin and SJW , both activators of the human pregnane X receptor ( O75469 ) , induced CYP3A in both liver and intestine but induced P-gp only in intestine , whereas Q15149 , an activator of murine O75469 , did not induce P-gp in any tissue . DB01045 disposition was evaluated , and hepatic exposure to rifampin was comparable to intestine ; in contrast , brain concentrations were low . Overall , these observations demonstrate that P-gp induction in vivo is tissue-specific ; furthermore , there is a disconnect between P-gp induction and CYP3A induction that is tissue- and inducer-dependent , suggesting that O75469 activation alone is insufficient for P-gp induction in vivo . Tissue-specific factors and inducer pharmacokinetic/pharmacodynamic properties may underlie these observations . P05231 and MYC collaborate in plasma cell tumor formation in mice . P05231 ( P05231 ) plays a critical role in the natural history of human plasma cell neoplasms ( PCNs ) , such as plasma cell myeloma and plasmacytoma ( PCT ) . P05231 is also at the center of neoplastic plasma cell transformation in BALB/c ( C ) mice carrying a transgene , H2-L(d)- P05231 , that encodes human P05231 under control of the major histocompatibility complex H2-L(d) promoter : strain C.H2-L(d)- P05231 . These mice are prone to PCT , but tumor development is incomplete with long latencies ( approximately 40 % PCT at 12 months of age ) . To generate a more robust mouse model of P05231 -dependent Q15149 , we intercrossed strain C.H2-L(d)- P05231 with strains C.iMyc(Emu) or C.iMyc(Calpha) , 2 interrelated gene-insertion models of the chromosomal T(12;15) translocation causing deregulated expression of Myc in mouse PCT . Deregulation of MYC is also a prominent feature of human Q15149 . We found that double-transgenic C.H2-L(d)- P05231 /iMyc(Emu) and C.H2-L(d)- P05231 /iMyc(Calpha) mice develop PCT with full penetrance ( 100 % tumor incidence ) and short latencies ( 3-6 months ) . The mouse tumors mimic molecular hallmarks of their human tumor counterparts , including elevated P05231 /Stat3/Bcl-X(L) signaling . The newly developed mouse strains may provide a good preclinical research tool for the design and testing of new approaches to target P05231 in treatment and prevention of human PCNs . Evidence of drug-drug interactions through uptake and efflux transport systems in rat hepatocytes : implications for cellular concentrations of competing drugs . For drugs with hepatobiliary transport across hepatocytes , the interplay between uptake and efflux transporters determines hepatic concentrations of drugs , but the evolution over time of these concentrations is difficult to measure in humans other than with magnetic resonance imaging contrast agents in the liver . DB00743 dimeglumine ( BOPTA ) is a contrast agent used in liver magnetic resonance imaging that enters into human hepatocytes through organic anion transporting polypeptides ( P46721 ) and exits unchanged into bile through the multiple resistance-associated protein 2 ( Q92887 ) . DB01045 ( Q9HBH0 ) is transported by the same membrane proteins and may compete with BOPTA for hepatic uptake . Simultaneous drug-drug interactions through uptake and efflux transport systems in hepatocytes according to the cellular concentrations of competing drugs were never investigated . In perfused rat liver preparations , we demonstrate how the drug-drug interactions through transporters determine cellular concentrations of the competing drugs BOPTA and Q9HBH0 , and we show that the cellular concentrations by modulating transport through membranes regulate the rat Oatp-Mrp2 interplay . Moreover , drug interactions through transporters change greatly over time . Impact of arsenite and its methylated metabolites on P09874 activity , P09874 gene expression and poly(ADP-ribosyl)ation in cultured human cells . The underlying mechanisms of arsenic carcinogenicity are still not fully understood . Mechanisms currently discussed include the induction of oxidative DNA damage and the interference with DNA repair pathways . Still unclear is the role of biomethylation , which has long been considered to be one major detoxification process . Methylated arsenicals have recently been shown to interfere with DNA repair in cellular and subcellular systems , but up to now no DNA repair protein has been identified being particular sensitive towards methylated arsenicals in cultured cells . Here we report that the trivalent methylated metabolites MMA(III) and P28067 (III) inhibit poly(ADP-ribosyl)ation in cultured human HeLa S3 cells at concentrations as low as 1nM , thereby showing for the first time an inactivation of an enzymatic reaction related to DNA repair by the trivalent methylated arsenicals at very low environmentally relevant concentrations . In contrast the pentavalent metabolites MMA(V) and P28067 (V) showed no such effects up to high micromolar concentrations . All investigated arsenicals did not alter gene expression of P09874 . However , all trivalent arsenicals were able to inhibit the activity of isolated P09874 , indicating that the observed decrease in poly(ADP-ribosyl)ation in cultures human cells , predominantly mediated by P09874 , is likely due to changes in the activity of P09874 . Since poly(ADP-ribosyl)ation plays a major role in DNA repair , cell cycle control and thus in the maintenance of genomic stability , these findings could in part explain DNA repair inhibition and the genotoxic and carcinogenic effects of arsenic . Q15149 isoform-dependent regulation of keratin-integrin alpha6beta4 anchorage via Ca2+/calmodulin . The detachment of epithelial cells from the basal matrix during wound healing and differentiation of keratinocytes requires the disassembly of the hemidesmosomal multiprotein adhesion complex . Integrin alpha6beta4-plectin interaction plays a major role in the formation of hemidesmosomes , and thus the mechanisms regulating this interaction should be critical also for the disassembly process . Here we show that a particular plectin isoform ( 1a ) interacts with the Ca(2+)-sensing protein calmodulin in a Ca(2+)-dependent manner . As a result of this interaction , binding of the hemidesmosome-associated plectin isoform 1a to integrin beta4 is substantially diminished . P62158 -binding inhibits also the interaction of plectin with F-actin . Further , we found that , during Ca(2+)-induced keratinocyte differentiation , plectin 1a is first relocated within the cell and later down-regulated , suggesting that Ca(2+) affects the fate of plectin 1a upon its release from hemidesmosomes . We propose a novel model for the disassembly of hemidesmosomes during keratinocyte differentiation , where both , binding of calmodulin to plectin 1a and phosphorylation of integrin beta4 by protein kinases , are required for disruption of the integrin alpha6beta4-plectin complex . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . DB06756 modulates age-related NF-kappaB by thiol-enhancing action . Depletion of glutathione levels and perturbations in redox status are considered to play a crucial role in aging and chronic inflammatory processes through the activation of redox sensitive transcription factors , including nuclear factor-kappaB ( NF-kappaB ) . In the current study , we assessed the regulatory action of dietary betaine in the suppression of NF-kappaB by comparing kidney tissue from old , betaine-supplemented rats or non-betaine-supplemented rats ( age 21 months ) and 7 month-old rats . In addition , cultured P29320 293T cells were utilized for the molecular assessment of betaine 's restorative ability of redox status when treating cells with potent glutathione ( DB00143 ) -depleting agents . Results showed that in old rats a short-term feeding ( 10 d ) with betaine attenuated the age-related decrease in thiol levels , increase in reactive species and TNFalpha expression via NF-kappaB activation , compared to the young controls . These findings were verified in the cell-cultured system . Further investigations found that redox imbalance due to thiol depletion caused increased NF-kappaB activation , and cyclooxygenase ( P36551 ) -2 and TNFalpha levels , both of which were suppressed by betaine treatment . Based on both in vivo and in vitro data , we concluded that betaine exerts its efficacy by maintaining thiol status in the regulation of P35354 and TNFalpha via NF-kappaB activation during aging . Cell-specific RNA interference by peptide-inhibited-peptidase-activated siRNAs . The use of chemically-synthesized short interfering RNAs ( siRNAs ) is the key method of choice to manipulate gene expression in mammalian cell cultures and in vivo . Several previous studies have aimed at inducing cell-specific RNA interference ( RNAi ) in order to use siRNA molecules as therapeutic reagents . Here , we used peptide-inhibited siRNAs that were activated after cleavage by cell-specific peptidases . We show that siRNAs with bound peptide at the antisense strand could be activated in target cells and were able to induce RNAi in a cell-specific manner . Green Fluorescent Protein ( GFP ) and Signal Transducer and Activator of Transcription ( P35610 ) -3 gene expression were selectively reduced in a JEG-3 human choriocarcinoma cell line expressing the activating enzyme caspase-4 , whereas the effect was absent in P29320 cells which lacked the enzyme . In JEG-3 cells , reduction of P40763 gene expression by conventional and peptide-inhibited siRNA led to a decrease in cell proliferation . This suggests that peptide-inhibited siRNAs provide improved cell specificity and offers new opportunities for their therapeutic use . Regulatory impact factors : unraveling the transcriptional regulation of complex traits from expression data . MOTIVATION : Although transcription factors ( TF ) play a central regulatory role , their detection from expression data is limited due to their low , and often sparse , expression . In order to fill this gap , we propose a regulatory impact factor ( Q9HBH0 ) metric to identify critical TF from gene expression data . RESULTS : To substantiate the generality of Q9HBH0 , we explore a set of experiments spanning a wide range of scenarios including breast cancer survival , fat , gonads and sex differentiation . We show that the strength of Q9HBH0 lies in its ability to simultaneously integrate three sources of information into a single measure : ( i ) the change in correlation existing between the TF and the differentially expressed ( DE ) genes ; ( ii ) the amount of differential expression of DE genes ; and ( iii ) the abundance of DE genes . As a result , Q9HBH0 analysis assigns an extreme score to those TF that are consistently most differentially co-expressed with the highly abundant and highly DE genes ( RIF1 ) , and to those TF with the most altered ability to predict the abundance of DE genes ( RIF2 ) . We show that Q9HBH0 analysis alone recovers well-known experimentally validated TF for the processes studied . The TF identified confirm the importance of Q07869 signaling in adipose development and the importance of transduction of estrogen signals in breast cancer survival and sexual differentiation . We argue that Q9HBH0 has universal applicability , and advocate its use as a promising hypotheses generating tool for the systematic identification of novel TF not yet documented as critical . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Photodynamic therapy with hypericin induces vascular damage and apoptosis in the Q9HBH0 -1 mouse tumor model . Hypericin , a polycyclic quinone obtained from plants of the genus Hypericum , has been proven to be a potent photosensitizer . The mechanism of tumor eradication and mode of cell death induced by in vivo photodynamic therapy ( PDT ) with hypericin were investigated in the present study using 2 therapeutic protocols . Q9HBH0 -1 tumors were exposed to laser light at either 0.5 hr or 6 hr after hypericin administration ( 5 mg/kg , i.v. ) . A significant reduction in tumor perfusion , as determined by the retention of fluorescein in the tumor tissue , was detected immediately after both PDT treatments . Further decrease in tumor perfusion was observed in the hours after treatment . The re-establishment of tumor perfusion , however , occurred 24 hr after 6 hr-interval PDT , but not after 0.5 hr-interval PDT . The kinetics of tumor cell survival estimated by the in vivo/in vitro clonogenic assay revealed no or limited cell death when tumors were explanted immediately after irradiation , whereas a delayed but progressive cell death was detected when tumors remained in situ after both PDT treatments . The detection of nucleosomal DNA fragmentation by agarose gel electrophoresis or TUNEL assay and the assessment of cell morphology by light microscopy indicated that apoptosis was the most prominent tumor response to hypericin-mediated PDT . Furthermore , immunohistochemical analysis of the tumor tissue showed an increased expression of both Fas and P48023 after irradiation , suggesting that this cell death pathway might contribute to the overall PDT-induced apoptotic response . In conclusion , our results demonstrate that apoptosis , likely occurring as a result of vascular damage , is responsible for the tumor eradication by PDT with hypericin in this tumor model . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . A novel role of transforming growth factor beta1 in transcriptional repression of human cholesterol 7alpha-hydroxylase gene . BACKGROUND & AIMS : Inhibition of cholesterol 7alpha-hydroxylase ( P22680 ) by bile acids and inflammatory cytokines provides an important mechanism to protect hepatocytes from bile acid toxicity during cholestasis . Transforming growth factor beta1 ( TGFbeta1 ) released by hepatic stellate cells during chronic liver injury plays a critical role in liver inflammation and fibrogenesis . The objective of this study is to investigate the role of TGFbeta1 in hepatic bile acid synthesis . METHODS : mRNA expressions in primary human hepatocytes and HepG2 cells were measured by quantitative real-time polymerase chain reaction . Reporter assay , glutathione-S-transferase pull-down assay , adenovirus-mediated gene transduction , and chromatin immunoprecipitation assay were used to study the mechanism of TGFbeta1 regulation of P22680 gene transcription . RESULTS : TGFbeta1 inhibited the mRNA expression of P22680 and bile acid synthesis in HepG2 cells and primary human hepatocytes . Mothers against decapentaplegic homolog ( P84022 ) inhibited both P22680 promoter activity and mRNA expression by inhibiting DNA-binding activity of hepatocyte nuclear factor 4alpha ( HNF4alpha ) . The histone deacetylase ( HDAC ) inhibitor Tricostatin A partially blocked the TGFbeta1 inhibition of P22680 mRNA expression , whereas TGFbeta1 decreased histone 3 acetylation in the P22680 chromatin . TGFbeta1 treatment and adenovirus P84022 reduced HNF4alpha binding but increased the recruitment of P84022 , Q13547 , and a repressor mSin3A to the P22680 chromatin . CONCLUSIONS : This study provides the first evidence that TGFbeta1 represses P22680 gene transcription in human hepatocytes by a mechanism involving P84022 -dependent inhibition of HNF4alpha and HDAC remodeling of P22680 chromatin . The TGFbeta1/ P84022 signaling may reduce bile acid synthesis in the liver and prevent hepatocyte injury in cholestatic liver disease . DB01045 -independent interactions between the pregnane X receptor ligand binding domain and peptide fragments of coactivator and corepressor proteins . The pregnane X receptor ( O75469 ) , a member of the nuclear receptor superfamily , regulates the expression of drug-metabolizing enzymes in a ligand-dependent manner . The conventional view of nuclear receptor action is that ligand binding enhances the receptor 's affinity for coactivator proteins , while decreasing its affinity for corepressors . To date , however , no known rigorous biophysical studies have been conducted to investigate the interaction among O75469 , its coregulators , and ligands . In this work , steady-state total internal reflection fluorescence microscopy ( TIRFM ) and total internal reflection with fluorescence recovery after photobleaching were used to measure the thermodynamics and kinetics of the interaction between the O75469 ligand binding domain and a peptide fragment of the steroid receptor coactivator-1 ( Q15788 ) in the presence and absence of the established O75469 agonist , rifampicin . Equilibrium dissociation and dissociation rate constants of ~5 μM and ~2 s(-1) , respectively , were obtained in the presence and absence of rifampicin , indicating that the ligand does not enhance the affinity of the O75469 and Q15788 fragments . Additionally , TIRFM was used to examine the interaction between O75469 and a peptide fragment of the corepressor protein , the silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) . An equilibrium dissociation constant of ~70 μM was obtained for Q9Y618 in the presence and absence of rifampicin . These results strongly suggest that the mechanism of ligand-dependent activation in O75469 differs significantly from that seen in many other nuclear receptors . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 -binding cassette transporter A1 ( O95477 ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 directly binds calmodulin in a Ca(2+)-dependent manner . The cytoplasmic loop of O95477 contains a typical calmodulin binding sequence of 1-5-8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5-8-14 motif abolished this interaction . This motif is located near the O95477 Pro- DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin/Ca(2+) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 and decreased O95477 protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 in the presence of Ca(2+) and increases the generation of high-density lipoprotein . P26651 -dependent post-transcriptional regulation of inflammatory cytokine mRNA expression by apolipoprotein A-I : role of DB00171 -binding membrane cassette transporter A1 and signal transducer and activator of transcription 3 . Atherosclerosis is an inflammatory disease characterized by the accumulation of macrophages in the arterial intima . The activated macrophages secreted more pro-inflammatory cytokines , such as tumor necrosis factor ( P01375 ) -α , which promote the development of the disease . P02647 ( apoA-I ) , the major component of high density lipoprotein , is involved in reverse cholesterol transport of lipid metabolism . Recently , it has been found that apoA-I suppresses inflammation via repression of inflammatory cytokine expression ; the mechanisms of the apoA-I-suppressive action , however , are not yet well characterized . In this study , we have for the first time found that apoA-I suppresses the expression of some inflammatory cytokines induced by lipopolysaccharide via a specific post-transcriptional regulation process , namely mRNA destabilization , in macrophages . Our further studies have also shown that AU-rich elements in the 3'-untranslated region of P01375 -α mRNA are responsive to the apoA-I-mediated mRNA destabilization . The apoA-I-induced inflammatory cytokine mRNA destabilization was associated with increased expression of mRNA-destabilizing protein tristetraprolin through a O60674 / P40763 signaling pathway-dependent manner . When blocking interaction of apoA-I with DB00171 -binding membrane cassette transporter A1 ( O95477 ) , a major receptor for apoA-I in macrophages , it would almost totally abolish the effect of apoA-I on tristetraprolin expression . These results present not only a novel mechanism for the apoA-I-mediated inflammation suppression in macrophages but also provide new insights for developing strategies for modulating vascular inflammation and atherosclerosis . Expression patterns and role of prostaglandin-endoperoxide synthases , prostaglandin E synthases , prostacyclin synthase , prostacyclin receptor , peroxisome proliferator-activated receptor delta and retinoid x receptor alpha in rat endometrium during artificially-induced decidualization . To determine if changes in endometrial expression of the enzymes and receptors involved in prostaglandin ( PG ) synthesis and action might provide insights into the PGs involved in the initiation of decidualization , ovariectomized steroid-treated rats at the equivalent of day 5 of pseudopregnancy were given a deciduogenic stimulus and killed at various times up to 32 h thereafter . The expression of PG-endoperoxide synthases ( P23219 and P35354 ) , microsomal PGE synthases ( O14684 and Q9H7Z7 ) , cytosolic PGE synthase ( Q15185 ) , prostacyclin synthase ( Q16647 ) , prostacyclin receptor , peroxisome proliferator-activated receptor delta ( Q03181 ) and retinoid x receptor alpha ( P19793 ) in endometrium was assessed by semiquantitative RT-PCR , western blot analyses and immunohistochemistry . In addition , to determine which PG is involved in mediating decidualization , we compared the ability of PGE(2) , stable analogues of P06744 (2) , L165041 ( an agonist of Q03181 ) , and docasahexanoic acid ( an agonist of P19793 ) to increase endometrial vascular permeability ( EVP , an early event in decidualization ) , and decidualization when infused into the uterine horns of rats sensitized for the decidual cell reaction ( DCR ) . EVP was assessed by uterine concentrations of Evans blue 10 h after initiation of infusions . DCR was assessed by the uterine mass 5 days after the initiation of the infusions . Because enzymes associated with the synthesis of PGE(2) , including P35354 , are up-regulated in response to a deciduogenic stimulus and because PGE(2) was more effective than the P06744 (2) analogues and Q03181 and P19793 agonists in increasing EVP and inducing decidualization , we suggest that PGE(2) is most likely the PG involved in the initiation of decidualization in the rat . Pseudomonas aeruginosa pyocyanin causes airway goblet cell hyperplasia and metaplasia and mucus hypersecretion by inactivating the transcriptional factor FoxA2 . The redox-active exotoxin pyocyanin ( Q15149 ) can be recovered in 100 µM concentrations in the sputa of bronchiectasis patients chronically infected with Pseudomonas aeruginosa ( PA ) . However , the importance of Q15149 within bronchiectatic airways colonized by PA remains unrecognized . Recently , we have shown that Q15149 is required for chronic PA lung infection in mice , and that chronic instillation of Q15149 induces goblet cell hyperplasia ( P30793 ) , pulmonary fibrosis , emphysema and influx of immune cells in mouse airways . Many of these pathological features are strikingly similar to the mouse airways devoid of functional FoxA2 , a transcriptional repressor of P30793 and mucus biosynthesis . In this study , we postulate that Q15149 causes and exacerbates P30793 and mucus hypersecretion in bronchiectatic airways chronically infected by PA by inactivating FoxA2 . We demonstrate that Q15149 represses the expression of FoxA2 in mouse airways and in bronchial epithelial cells cultured at an air-liquid interface or conventionally , resulting in P30793 , increased Q9HC84 mucin gene expression and mucus hypersecretion . Immunohistochemical and inhibitor studies indicate that Q15149 upregulates the expression of Stat6 and P00533 , both of which in turn repress the expression of FoxA2 . These studies demonstrate that Q15149 induces P30793 and mucus hypersecretion by inactivating FoxA2 . Organic anion transporting polypeptide-C mediates arsenic uptake in P29320 -293 cells . Arsenic is an established human carcinogen . The role of aquaglyroporins ( AQPs ) in arsenic disposition was recently identified . In order to examine whether organic anion transporting polypeptide-C ( Q9Y6L6 ) also plays a role in arsenic transport , Q9Y6L6 cDNA was transfected into cells of a human embryonic kidney cell line ( P29320 -293 ) . Transfection increased uptake of the model Q9Y6L6 substrate , estradiol-17beta-D-glucuronide , by 10-fold . In addition , we measured uptake and cytotoxicity of arsenate , arsenite , monomethylarsonate(MMA(V)) , and dimethylarsinate ( P28067 (V) ) . Transfection of Q9Y6L6 increased uptake and cytotoxicity of arsenate and arsenite , but not of MMA(V) or P28067 (V) . DB01045 and taurocholic acid ( a substrate of Q9Y6L6 ) reversed the increased toxicity of arsenate and arsenite seen in Q9Y6L6 -transfected cells . The increase in uptake of inorganic arsenic was not as great as that of estradiol-17beta-D-glucuronide . Our results suggest that Q9Y6L6 can transport inorganic arsenic in a ( DB00143 ) -dependent manner . However , this may not be the major pathway for arsenic transport .

MH_train_1009

MH_train_1009

MH_train_1009

interacts_with DB00013?

multiple_choice

Modulation of mitogen-activated protein kinase cascades by differentiation-1 protein : acquired drug resistance of hormone independent prostate cancer cells . PURPOSE : The inhibitor of differentiation-1 protein ( Id-1 ) is over expressed in multidrug resistance prostate cancer cells . We determined the effect of Id-1 expression and its underlying pathways on the development of multidrug resistance in prostate cancer . MATERIALS AND METHODS : P01008 cells were transfected with the Id-1 gene or a blank vector . Id-1 mRNA expression was determined by reverse transcriptase-polymerase chain reaction and Id-1 protein content was detected by immunoblot and flow cytometry . Cellular cytotoxicity was determined by MTT ( microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ) assay ( Sigma Chemical Co. , St. Louis , Missouri ) . The activation and expression of mitogen-activated protein kinase ( MAPK ) were measured by transactivation assay and Western blotting , respectively . RESULTS : Id-1 overproduction drove P01008 cells to become resistant to chemotherapeutic agents but did not induce mdr-1 gene expression . The p38MAPK and c-jun N-terminal kinase ( JNK ) pathways were suppressed , which correlated with increased Id-1 expression . No significant change in extracellular signal-regulated kinase ( P29323 ) activation was observed in Id-1 transfectants compared with that of P01008 or vector control . Treatment of Id-1 expressing cells with p38MAPK and JNK inhibitors resulted in decreased doxorubicin induced apoptosis . In contrast , Id-1 expressing cells treated with P29323 inhibitor made cells more sensitive to drug induced apoptosis . CONCLUSIONS : Up-regulation of Id-1 was found in prostate cancer multidrug resistant cells . Sustained P29323 activation , and JNK and p38MAPK inhibition by Id-1 in cells may confer drug resistance . These changes in MAPKs could be a mechanism for the acquisition of multidrug resistance in prostate cancer . Lipid raft compartmentalization of urokinase receptor signaling in human neutrophils . DB00013 plasminogen activator ( uPA ) receptors ( Q03405 ) can be engaged for activation signaling either by aggregation or by binding exogenous uPA . These signaling mechanisms require Q03405 to associate with two distinct adhesion proteins , P14151 and complement receptor 3 ( CR3 ) , respectively . Q03405 contains a glycosylphosphatidylinositol anchor , suggesting that it is concentrated within glycosphingolipid-enriched microdomains , or " lipid rafts " . This study was undertaken to determine the extent to which Q03405 -mediated signaling is compartmentalized to lipid rafts . Human neutrophil Q03405 was cross-linked or stimulated with uPA after pretreatment with the lipid raft-disrupting agents , methyl-beta-cyclodextrin or filipin III . Both agents suppressed increases in intracellular Ca(2+) concentrations ( [Ca(2+)](i) ) triggered by cross-linking , but did not affect [ Ca(2+) ] (i) in response to uPA . Neutrophil membranes were separated into lipid raft and non-raft fractions , revealing the presence of Q03405 and P14151 , but the virtual absence of CR3 alpha chain in lipid rafts , either constitutively or in response to Q03405 aggregation . Fluorescence resonance energy transfer experiments confirmed close proximity of a lipid raft marker to both Q03405 and P14151 , but not CR3 . We conclude that Q03405 can engage distinct signaling pathways involving different partner proteins that are functionally and physically segregated from one another in both lipid raft and non-raft domains of the plasma membrane . Molecular imaging of pancreatic cancer in an animal model using targeted multifunctional nanoparticles . BACKGROUND & AIMS : Identification of a ligand/receptor system that enables functionalized nanoparticles to efficiently target pancreatic cancer holds great promise for the development of novel approaches for the detection and treatment of pancreatic cancer . DB00013 plasminogen activator receptor ( Q03405 ) , a cellular receptor that is highly expressed in pancreatic cancer and tumor stromal cells , is an excellent surface molecule for receptor-targeted imaging of pancreatic cancer using multifunctional nanoparticles . METHODS : The Q03405 -targeted dual-modality molecular imaging nanoparticle probe is designed and prepared by conjugating a near-infrared dye-labeled amino-terminal fragment of the receptor binding domain of urokinase plasminogen activator to the surface of functionalized magnetic iron oxide nanoparticles . RESULTS : We have shown that the systemic delivery of Q03405 -targeted nanoparticles leads to their selective accumulation within tumors of orthotopically xenografted human pancreatic cancer in nude mice . The Q03405 -targeted nanoparticle probe binds to and is subsequently internalized by Q03405 -expressing tumor cells and tumor-associated stromal cells , which facilitates the intratumoral distribution of the nanoparticles and increases the amount and retention of the nanoparticles in a tumor mass . Imaging properties of the nanoparticles enable in vivo optical and magnetic resonance imaging of Q03405 -elevated pancreatic cancer lesions . CONCLUSIONS : Targeting Q03405 using biodegradable multifunctional nanoparticles allows for the selective delivery of the nanoparticles into primary and metastatic pancreatic cancer lesions . This novel receptor-targeted nanoparticle is a potential molecular imaging agent for the detection of pancreatic cancer . DB00013 receptor is associated with the components of the P23458 / P42224 signaling pathway and leads to activation of this pathway upon receptor clustering in the human kidney epithelial tumor cell line Q9H4E5 -598 . The urokinase-type plasminogen activator ( uPA ) binds to cells via a specific receptor attached to the plasma membrane by a glycosylphosphatidylinositol ( P06744 ) anchor . Despite the lack of a transmembrane domain , the urokinase receptor ( Q03405 ) is capable of transducing extracellular signals affecting growth , migration , and adhesion . Several DB00135 kinases of the src family as well as beta1 , beta2 , and beta3 integrins were found to be associated with the Q03405 . We found that in the human kidney epithelial line Q9H4E5 -598 , also components of the P23458 / P42224 signal transduction pathway including P40189 , are associated with Q03405 as revealed by coimmunoprecipitation and are co-localized in caveolae . Upon clustering of uPA. Q03405 complex by a monoclonal antibody , P23458 associates with Q03405 , which in turn leads to P42224 phosphorylation , dimerization , specific binding to DNA , and gene activation . To prove the dependence of P42224 activation on the Q03405 , Q9H4E5 -598 cells were treated with sense and antisense Q03405 oligonucleotides . In antisense-treated cells in which Q03405 expression was reduced to less then one third , activation of P42224 by the clustering antibody was abolished while P42224 activation by interferon-gamma was unaffected . Therefore , in this cell line , uPA. Q03405 also utilizes the P23458 / P42224 pathway for signaling , and P40189 might be the transmembrane adapter for this signal transduction pathway . Expression of urokinase , plasminogen activator inhibitors and urokinase receptor in pregnant rhesus monkey uterus during early placentation . We have investigated plasmin mediated proteolysis associated with trophoblast invasion during early stages of pregnancy in the rhesus monkey . In situ hybridization and immunocytochemical localization were used to define the cellular and tissue distribution of urokinase plasminogen activator ( uPA ) , plasminogen activator inhibitor type 1 ( P05121 ) and 2 ( P05120 ) and urokinase receptor in early monkey placenta and uterus . Our results indicate : ( 1 ) uPA is expressed in proliferating and invasive cytotrophoblast located in chorionic villi as well as in extravillous trophoblast associated with uterine arterioles . This raises the possibility that urokinase may play an important role in trophoblast invasion . ( 2 ) P05121 mRNA is specifically localized in two areas where invasive trophoblast cells encounter maternal tissue directly . The extravillous cytotrophoblast cells at the maternofetal junction express P05121 mRNA . The invasive endovascular trophoblast cells within the uterine arterioles also express P05121 mRNA . The location sensitive expression of P05121 mRNA at the maternofetal junction may imply a protective function of this protease inhibitor that might be induced through interaction with decidual cells . ( 3 ) DB00013 receptor antigen has also been found at the maternofetal junction and in endovascular trophoblast cells of the invaded maternal blood vessel . ( 4 ) P05120 immunoreactivity is found in association with cytotrophoblast cells in anchoring choronic villi suggesting its association with early placentation . In conclusion , we propose that the plasmin/plasminogen activator system may not only regulate extracellular matrix degradation , but also modify migration and invasive behaviour of extravillous trophoblast cells , during early placentation . DB00013 plasminogen activator and urokinase plasminogen activator receptor in breast cancer . DB00013 plasminogen activator ( uPA ) is a serine protease involved in cancer invasion and metastasis . uPA mediates its action while attached to a membrane-bound receptor ( Q03405 ) . In this investigation we show that Q03405 levels correlate with uPA levels in human breast cancers . Q03405 levels , however , do not correlate with other components of the plasminogen activator system such as tissue-type plasminogen activator ( t-PA ) , P05121 -I or P05120 . In addition , Q03405 levels showed no correlation with tumor size , axillary-node status or estrogen-receptor status . On the basis of an optimum cut-off point , patients with breast cancers containing high levels of Q03405 had a worse prognosis than patients with low levels of the receptor . However , as a prognostic marker in breast cancer , Q03405 was not as strong as uPA . Our results are consistent with data from model systems suggesting that both uPA and Q03405 are necessary for metastasis . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . uPA receptor expression in benign and malignant thyroid tumors . BACKGROUND : DB00013 type plasminogen activator receptor ( Q03405 ) plays an important role in cancer invasion and metastasis . However , the Q03405 expression has been rarely investigated in thyroid carcinomas . The aim of this study was to evaluate the clinical relevance of Q03405 in thyroid tumors . MATERIALS AND METHODS : Samples included 53 benign tumors ( follicular adenoma 34 , Graves ' disease 8 , adenomatous goiter 7 and others 4 ) and 62 cancers ( papillary thyroid cancer ( PTC ) 47 , follicular TC ( FTC ) 5 , medullary TC ( P04629 ) 5 and anaplastic TC ( ATC ) 5 ) . Q03405 expression was prospectively investigated with a labeled streptavidin-biotin method using an anti- Q03405 monoclonal antibody . Patients were classified into a low- and high-staining group according to the percentage of positive cells ( cut-off value=10 % ) . RESULTS : Q03405 was more strongly expressed in thyroid cancers ( 35.5 % ) than benign tumors ( 7.5 % ) . FTC had a significantly higher Q03405 expression compared to follicular adenoma ( p < 0.01 ) . The positivity of Q03405 was as follows : PTC 36.2 % , FTC 60 % , P04629 0 % and ATC 40 % . In PTC , high Q03405 expression was associated with poorly-differentiated PTC ( p < 0.01 ) while had a trend to develop more distant metastases than those with low Q03405 expression ( p=0.17 , by the Kaplan-Meier method ) . CONCLUSION : This study has shown that Q03405 expression might be useful for the discrimination between FTC and follicular adenoma and could possibly be used as a prognostic factor in PTC . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . P00747 activator urokinase expression reveals P50591 responsiveness and supports fractional survival of cancer cells . P01375 -related apoptosis-inducing ligand ( P50591 / P50591 /Apo2L ) holds promise for cancer therapy as it induces apoptosis in a large variety of cancer cells while exerting negligible toxicity in normal ones . However , P50591 can also induce proliferative and migratory signaling in cancer cells resistant to apoptosis induced by this cytokine . In that regard , the molecular mechanisms underlying the tumor selectivity of P50591 and those balancing apoptosis versus survival remain largely elusive . We show here that high mRNA levels of P00749 , which encodes urokinase plasminogen activator ( uPA ) , are characteristic of cancer cells with functional P50591 signaling . Notably , decreasing uPA levels sensitized cancer cells to P50591 , leading to markedly increased apoptosis . Mechanistic analyses revealed three molecular events taking place in uPA-depleted cells : reduced basal P27361 /2 prosurvival signaling , decreased preligand decoy receptor 2 ( Q9UBN6 ) -death receptor 5 ( DR5 ) interaction and attenuated recruitment of Q9UBN6 to the death-inducing signaling complex upon P50591 challenge . These phenomena were accompanied by increased Q13158 and procaspase-8 recruitment and processing , thus guiding cells toward a caspase-dependent cell death that is largely independent of the intrinsic apoptosis pathway . Collectively , our results unveil P00749 mRNA levels as marker for the identification of P50591 -responsive tumor cells and highlight a key role of uPA signaling in ' apoptosis versus survival ' decision-making processes upon P50591 challenge . (Pro)renin promotes fibrosis gene expression in P29320 cells through a Nox4-dependent mechanism . The (pro)renin receptor ( PRR ) has recently been demonstrated to bind equally well renin and its precursor , prorenin , leading to a similar intracellular signaling independent of angiotensin II . In this study , we report that human embryonic kidney cells ( P29320 ) exposed to renin or prorenin for 24 h in the presence of a blocking concentration of the angtiotensin-converting enzyme inhibitor perindoprilate increased superoxide anion production as measured by luminescence ( lucigenin ) and electron spin resonance spectroscopy ( hydroxylamine radical transition ) . Also , both renin and prorenin increased Nox4 expression while Nox2 , p47(phox) , and p67(phox) remained unchanged . In an investigation of the effects of renin and prorenin on fibrosis genes , it appeared that both proteins stimulated transforming growth factor-β ( TGF-β ) , fibronectin , and plasminogen activator inhibitor type 1 ( P05121 ) expression and therefore participated to an overall switch toward a profibrotic state of the kidney cells . When the cells were transfected with a siRNA targeting the PRR , Nox4 expression was efficiently prevented as well as the increase in superoxide production , TGF-β , fibronectin , and P05121 . Finally , we demonstrated that transfection of the cells with a Nox4-specific small interfering ( si ) RNA also prevented fibrosis gene expression following treatment with renin or prorenin . The results demonstrate that renin and prorenin , through their specific membrane receptor and independently of angiotensin II , promote fibrosis gene expression via a Nox4-dependent mechanism . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Abnormalities of the urokinase system in colonic crypt cells from patients with ulcerative colitis . : DB00013 [ urokinase-type plasminogen activator ( u-PA ) ] is potentially involved in the control of epithelial cell adhesion and repair processes . This study aimed to identify abnormalities in the responses in vitro of the u-PA system in colonic crypt cells in ulcerative colitis and Crohn 's disease . Initial experiments demonstrated expression and transcription of receptors for u-PA ( u-PAR ) by colonic crypt cells . Differences across disease groups were found in results from cells isolated from noninflamed segments of colon . Supernatant and cell-associated u-PA activities in ulcerative colitis were significantly greater than normal , and supernatant u-PA activity was greater than in Crohn 's disease . These differences were only partly explained by increased u-PA secretion compared with normal and lower plasminogen activator inhibitor-1 ( P05121 ) secretion than in Crohn 's disease . The effect of 20-h exposure to butyrate ( 1 mmol/ L ) was similar across disease groups except for a failure to stimulate P05121 secretion in ulcerative colitis and Crohn 's disease and to suppress u-PAR expression in Crohn 's disease . In ulcerative colitis , colonic crypt cells from inflamed mucosa exhibited significantly lower cell-associated and supernatant u-PA activities and altered butyrate-mediated responses in cell-associated u-PA activity and u-PAR expression than cells from uninflamed mucosa . Perturbed colonic crypt cell responses of the u-PA system in vitro in ulcerative colitis suggest the presence of a primary diffuse abnormality in the colonic epithelium . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract . [ Plasma exchange with very low molecular weight heparin CY 222. Biological profile and therapeutic value ] . In a clinical , between-patient study we investigated the effects of a VLMW DB01109 fragment ( CY 222 ) versus standard heparin ( SH ) in plasma exchanges ( n = 10 ) on coagulation factors ( CF ) . DB09222 ( FGN ) , II , V , VIIF + X , IX , XI , XII , VIIIc , VIIIRag , VIIIvwf and P01008 , ProtC , ProtS , P00747 ( Q9UQ90 ) , Activated P13726 time ( APTT ) , P00734 time ( PT ) , Thrombin time ( TT ) , anti factor Xa ( Axa ) , DDimer , Platelet count . DB01109 was administered as a bolus and by infusion during the session , CY 222 as a bolus dose only ; 1 to 1.5 plasma volume was exchanged with substitution by 5 % albumin . Results ( mean , s.d. ) at the end of the session ( End ) and 4 hours later ( DB00451 ) were analyzed and showed no differences between groups ( with CY 222 and with SH ) for technical and clinical findings . Biologically , CF were similar in both groups except for Factor XII levels at the end of the session , and Factors II , V , XII at DB00451 . Prolonged APTT in all samples appear related to low FGN . Significant differences in Axa activities were found for each treatment when compared with its own standard , suggesting different ranges of activities of both drugs . Changes in D-dimer levels differed during the session and four hours after the session with the drug tested , and could be related to their mode of administration . Clinical efficiency and tolerance were excellent both with CY 222 and SH . The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy . Concomitant expression of urokinase type plasminogen activator ( uPA ) and its surface receptor ( Q03405 ) has been shown to correlate strongly with a more invasive tumor cell phenotype . A highly malignant human epidermoid carcinoma cell line ( HEp3 ) was transfected with a vector capable of expressing an antisense transcript complementary to 300 bases of the 5' end of Q03405 , including the ATG codon . Six stably transfected antisense ( AS-2 , 3 , 5 , 9 , 10 , 12 ) and eight control clones were characterized . All clones produced high levels of uPA activity . Examination of collagenase production and doubling time showed that all of the clones tested produced similar activities . The antisense clones showed a 20-74 % reduction in the Q03405 sites ; the Q03405 mRNA level was also reduced . A test of the invasive ability of all clones in a modified chorioallantoic membrane ( P62158 ) showed that invasiveness of the antisense-inhibited clones was directly proportional to the density of surface Q03405 . The AS-2 clone , which expressed the lowest number of uPARs showed a significantly reduced level of invasion . The invasiveness of additional AS-inhibited clones was also reduced . Seven control and four AS-inhibited clones were tested for tumorigenicity on CAMs of chick embryos . Inoculation of control cells produced large tumors , while the As clones were non-tumorigenic . AS-2 did not produce tumors even if kept in vivo for up to 10 weeks. ( ABSTRACT TRUNCATED AT 250 WORDS ) Epithelial and stromal cell urokinase plasminogen activator receptor expression differentially correlates with survival in rectal cancer stages B and C patients . DB00013 plasminogen activator receptor ( Q03405 ) has been proposed as a potential prognostic factor for colorectal cancer ( CRC ) patient survival . However , CRC Q03405 expression remains controversial , especially regarding cell types where Q03405 is overexpressed ( e.g. , epithelium ( uPARE ) or stroma-associated cells ( uPARS ) ) and associated prognostic relevance . In this study , two epitope-specific anti- Q03405 monoclonal antibodies ( MAbs ) could discriminate expression of uPARE from uPARS and were used to examine this association with survival of stages B and C rectal cancer ( RC ) patients . Using immunohistochemistry , MAbs # 3937 and R4 were used to discriminate uPARE from uPARS respectively in the central and invasive frontal regions of 170 stage B and 179 stage C RC specimens . Kaplan-Meier and Cox regression analyses were used to determine association with survival . Q03405 expression occurred in both epithelial and stromal compartments with differential expression observed in many cases , indicating uPARE and uPARS have different cellular roles . In the central and invasive frontal regions , uPARE was adversely associated with overall stage B survival ( HR = 1.9 ; p = 0.014 and HR = 1.5 ; p = 0.031 , respectively ) reproducing results from previous studies . uPARS at the invasive front was associated with longer stage C survival ( HR = 0.6 ; p = 0.007 ) , reflecting studies demonstrating that macrophage peritumoural accumulation is associated with longer survival . This study demonstrates that different Q03405 epitopes should be considered as being expressed on different cell types during tumour progression and at different stages in RC . Understanding how uPARE and uPARS expression affects survival is anticipated to be a useful clinical prognostic marker of stages B and C RC . DB00013 receptor up-regulation in head and neck squamous cell carcinoma . BACKGROUND : P00749 is important for matrix degradation and motility of cancer cells . For effective invasion , urokinase has to be associated with its cell surface receptor.(1) METHODS : We analyzed 33 head and neck squamous cell carcinomas ( hnSCC ) and 14 mucosal tissue samples for the expression of urokinase receptor using Northern hybridization and correlated expression levels to clinical and histopathologic data . DB00013 expression was determined by fibrin zymography . RESULTS : The expression of urokinase receptor is significantly increased in hnSCC compared with adjacent mucosa . Expression levels in primary tumors show no statistically significant correlations to T staging , metastasis , recurrence , or differentiation stage of the resected tumors . Furthermore , there was no correlation between urokinase and urokinase receptor expression levels in SCC samples . CONCLUSIONS : DB00013 receptor expression is increased in hnSCC , but it is not useful as a prognostic marker for the metastatic behavior of primary tumors . Comparison of our data with previously published reports is discussed . Leukocyte urokinase plasminogen activator receptor and PSGL1 play a role in endogenous arterial fibrinolysis . Fibrin is an integral component of arterial thrombi . Using a mouse model of arteriolar thrombosis , high-speed fluorescence microscopy reveals that , within minutes , the fibrin content of thrombi rapidly increases and then decreases . The decrease in fibrin coincides with leukocyte binding to the thrombi , a process mediated by the interaction of leukocyte P16109 glycoprotein ligand-1 ( Q14242 ) with P16109 on the surface of activated platelets . Because leukocytes possess urokinase-type plasminogen activator ( uPA ) activity , we used mice deficient in uPA or the uPA receptor ( Q03405 ) to explore the contribution of leukocyte-associated uPA to the loss of fibrin from these thrombi . Fibrin loss in both uPA-deficient mice and Q03405 -deficient mice was reduced compared with that in wild-type controls . Transfusion of leukocytes from wild-type mice into Q03405 -deficient mice restored fibrin loss to levels similar to that in wild-type mice . In contrast , transfusion of leukocytes from mice deficient in Q03405 or Q14242 did not enhance fibrin loss . Thus , fibrin loss from microarteriolar thrombi is mediated , at least in part , by leukocyte-associated uPA in a process that requires leukocyte Q03405 and Q14242 . Development of peptidomimetic ligands of Pro- DB00149 - DB00145 -NH(2) as allosteric modulators of the dopamine D(2) receptor . A variety of stable , small-molecule peptidomimetic ligands have been developed to elucidate the mechanism by which the neuropeptide Pro- DB00149 - DB00145 -NH(2) ( P00747 ) modulates dopaminergic neurotransmission . Photoaffinity labeling ligands based upon P00747 peptidomimetics have been used to establish that P00747 binds to the P14416 at a site that is different from the orthosteric site , thus making P00747 and its peptidomimetics allosteric modulators of the dopamine receptor . Through the design , synthesis and pharmacological evaluation of conformationally constrained peptidomimetics containing lactam , bicyclic , and spiro-bicyclic scaffolds , support was provided for the hypothesis that the bioactive conformation of P00747 is a type II β-turn . In addition , studies with peptidomimetics designed to mimic either a type VI β-turn or polyproline II helix conformation yielded molecules that were able to modulate dopamine receptors because of their ability to place the carboxamide NH(2) pharmacophore in the same topological space as that seen in the type II β-turn . Extensive studies with the spiro-bicyclic P00747 peptidomimetics also established that both positive and negative modes of modulation were possible for the same series of peptidomimetics simply as a result of minor differences in the stereochemistry about the bridgehead carbon within the scaffold . This information was used to transform existing positive modulators into negative modulators , which demonstrated that small structural changes in the spiro-bicyclic dopamine receptor modulators are capable of causing major changes in the modulatory activity of P00747 peptidomimetics . P00747 activator inhibitor type-1 inhibits insulin signaling by competing with alphavbeta3 integrin for vitronectin binding . Functional cooperation between integrins and growth factor receptors has been reported for several systems , one of which is the modulation of insulin signaling by alphavbeta3 integrin . P00747 activator inhibitor type-1 ( P05121 ) , competes with alphavbeta3 integrin for vitronectin ( VN ) binding . Here we report that P05121 , in a VN-dependent manner , prevents the cooperation of alphavbeta3 integrin with insulin signaling in NIH3T3 fibroblasts , resulting in a decrease in insulin-induced protein kinase B ( P31749 ) phosphorylation , vascular endothelial growth factor ( P15692 ) expression and cell migration . P01308 -induced HUVEC migration and angiotube formation was also enhanced in the presence of VN and this enhancement is inhibited by P05121 . By using specific P05121 mutants with either VN binding or plasminogen activator ( PA ) inhibiting activities ablated , we have shown that the P05121 -mediated interference with insulin signaling occurs through its direct interaction with VN , and not through its PA neutralizing activity . Moreover , using cells deficient for uPA receptor ( Q03405 ) we have demonstrated that the inhibition of P05121 on insulin signaling is independent of Q03405 -VN binding . These results constitute the first demonstration of the interaction of P05121 with the insulin response . Protective effects of estradiol on P50591 -induced apoptosis in a human oligodendrocytic cell line : evidence for multiple sites of interactions . Demyelinating diseases are high impact neurological disorders . Steroids are regarded as protective molecules in the susceptibility to these diseases . Here , we studied the interactions between tumour necrosis factor-related apoptosis-inducing ligand ( P50591 ) , a potent proapoptotic molecule toxic to oligodendrocytes , and 17-beta-estradiol ( E-17-beta ) , in human oligodendrocytic Q03405 .13 cells . Exposure of cells to P50591 resulted in the upregulation of both death receptors DR4 and DR5 and apoptosis , as well as the activation of caspase-8 and -3 , increased phosphorylation of Jun-N-terminal kinase and p38 kinase , and the reduction of bcl-2 and bcl-xL proteins . P50591 -mediated Q03405 .13 cell apoptosis was abrogated by the dominant-negative form of the adaptor protein Q13158 and by caspase inhibitors . Preincubation with E-17-beta completely prevented both P50591 -induced DR4 and DR5 upregulation and apoptosis . Estrogen-induced cytoprotection was time and concentration dependent and reverted by antiestrogens . Estrogen treatment per se reduced kinase phosphorylation , and upregulated bcl-2 and bcl-xL proteins . In conclusion , our data show that the detrimental role of P50591 on oligodendrocytes can be effectively counteracted by estrogens , thus suggesting that the underlying molecular interactions can be of potential relevance in characterizing novel targets for therapy of demyelinating disorders . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Activated human neutrophils rapidly release the chemotactically active D2D3 form of the urokinase-type plasminogen activator receptor ( Q03405 /CD87 ) . The urokinase-type plasminogen activator receptor ( Q03405 /CD87 ) exists both in cell-bound and soluble forms . Neutrophils contain extensive intracellular pools of Q03405 that are translocated to the plasma membrane upon activation . In the present study , we investigated the ability of human neutrophils to shed Q03405 from cell surface following activation and addressed the possible involvement of the released receptor in the inflammatory response . We first observed that the spontaneous release of suPAR by resting neutrophils was strongly and rapidly ( within minutes ) enhanced by calcium ionophore ionomycin and to a lesser extent when cells were primed with P01375 and then stimulated with fMLP or P10145 . We demonstrated that suPAR is produced by resting and activated neutrophils predominantly as a truncated form devoid of N-terminal D1 domain ( D2D3 form ) that lacks P06744 anchor . Migration of formyl peptide receptor-like 1 ( P25090 ) -transfected human embryonic kidney ( P29320 ) 293 cells toward the supernatants harvested from activated neutrophils was significantly diminished when D2D3 form of suPAR was immunodepleted from the supernatants . We conclude that activated neutrophils release the chemotactically active D2D3 form of suPAR that acts as a ligand of P25090 . Interestingly , we present evidence that P80108 ( P80108 ) that has previously been shown to shed Q03405 in cancer cells is not involved in suPAR release from human neutrophils . We suggest that production of the chemotactically active D2D3 form of suPAR by activated human neutrophils in vivo could contribute to the recruitment of monocytes and other formyl peptide receptors-expressing cells to the sites of acute inflammation where neutrophil accumulation and activation occur . The role of urokinase-type plasminogen activator in aggressive tumor cell behavior . The correlation between urokinase-type plasminogen activator ( uPA ) expression and tumor cell invasion and metastasis has been well documented . DB00013 converts the zymogen plasminogen to plasmin , a trypsin-like enzyme with broad substrate specificities . Net uPA activity is determined not only by the amount of the enzyme itself , but also by its state of activation and the amount of specific plasminogen activator inhibitors ( PAIs ) present . Both uPA and its substrate , plasminogen , can bind to cells via specific membrane-associated receptors . Expression of uPA , uPA receptor ( Q03405 ) , and PAIs is regulated by growth factors , oncogenes , and other effector molecules . In the present review we discuss the interactions of uPA with its receptor , inhibitors , and substrate and how these interactions influence malignant behavior . We also review recent reports in which investigators have used anti-catalytic antibodies and/or gene transfection to demonstrate that uPA is directly involved in tumor cell invasion and metastasis . AZD1480 blocks growth and tumorigenesis of P07949 - activated thyroid cancer cell lines . Persistent P07949 activation is a frequent event in papillary thyroid carcinoma ( PTC ) and medullary thyroid carcinoma ( P04629 ) . In these cancers , P07949 activates the P29323 /MAPK , the PI3K/AKT/ P42345 and the JAK/ P40763 pathways . Here , we tested the efficacy of a P23458 /2- inhibitor , AZD1480 , in the in vitro and in vivo growth of thyroid cancer cell lines expressing oncogenic P07949 . Thyroid cancer cell lines harboring P07949 / Q13635 ( TPC-1 ) , P07949 M918T ( MZ-CRC1 ) and P07949 C634W ( TT ) alterations , as well as TPC-1 xenografts , were treated with JAK inhibitor , AZD1480 . This inhibitor led to growth inhibition and/or apoptosis of the thyroid cancer cell lines in vitro , as well as to tumor regression of TPC-1 xenografts , where it efficiently blocked P40763 activation in tumor and stromal cells . This inhibition was associated with decreased proliferation , decreased blood vessel density , coupled with increased necrosis . However , AZD1480 repressed the growth of P40763 - deficient TPC-1 cells in vitro and in vivo , demonstrating that its effects in this cell line were independent of P40763 in the tumor cells . In all cell lines , the JAK inhibitor reduced phospho-Y1062 P07949 levels , and P42345 effector phospho-S6 , while P23458 /2 downregulation by siRNA did not affect cell growth nor P07949 and S6 activation . In conclusion , AZD1480 effectively blocks proliferation and tumor growth of activated P07949 - thyroid cancer cell lines , likely through direct P07949 inhibition in cancer cells as well as by modulation of the microenvironment ( e.g. via JAK/phospho- P40763 inhibition in endothelial cells ) . Thus , AZD1480 should be considered as a therapeutic agent for the treatment of P07949 - activated thyroid cancers . DB00013 receptor surface expression regulates monocyte migration and is associated with accelerated atherosclerosis . BACKGROUND : The urokinase receptor ( Q03405 ) is a key regulator of pericellular proteolysis , cell adhesion and migration , all of which are fundamental processes in atherogenesis . We hypothesized that increased monocytic Q03405 expression in circulation is associated with the formation and development of atherosclerosis . METHODS : A total of 42 male apoE-/- mice were ramdonly divided into high-fat ( HF ) diet and normal diet ( n=21 per group ) . The percentage of Q03405 expressing monocytes ( PUEM ) and the expression of Q03405 within different types of atherosclerotic plaques were measured at an interval of 3 weeks from week 10 to week 16 . In vitro , Q03405 expression upon ox-LDL stimulation and the migration of monocytes were examined . RESULTS : PUEM in circulation was significantly higher in animals with HF diet compared with those having normal diet ( p < 0.03 ) . The augmented levels of PUEM were associated with body weight , visceral fat weight and numbers of Q03405 +macrophages within atherosclerotic lesions . Accumulation of Q03405 +macrophages increased with the progression of atherosclerosis . Monocytes upon ox-LDL stimulation exhibited an increased Q03405 expression and Q03405 antibody markedly suppressed monocyte migration induced by monocyte chemotactic protein-1 . Q03405 modulated monocyte migration was accelerated by uPA and was suppressed by amino terminal fragment of uPA dependent . CONCLUSIONS : Over-expression of Q03405 both in circulating monocytes and in atherosclerotic lesions is associated with the development of atherosclerotic plaques . Q03405 and its interaction with uPA may contribute to enhanced monocyte migration . Thus , Q03405 may be a novel target for prevention of uncontrolled monocyte recruitment in inflammatory atherogenic process . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Multiple roles of the candidate oncogene O75362 in ovarian epithelial neoplastic progression . The transcription factor O75362 is often amplified in ovarian cancer , but its role in neoplastic progression is unknown . We introduced O75362 -HA by adenoviral and retroviral infection into normal human ovarian surface epithelial cells ( OSE ) , i.e. , the source of ovarian cancer , and into SV40 Tag/tag expressing , p53/pRB-deficient OSE with extended but finite life spans ( IOSE ) . In OSE , O75362 -HA reduced cell-substratum adhesion and accelerated loss of senescent cells , but caused no obvious proneoplastic changes . In contrast , O75362 -HA transduction into IOSE yielded two permanent lines , I-80RZ and I-144RZ , which exhibited telomerase activity , stable telomere lengths , anchorage independence and reduced serum dependence , but were not tumorigenic in SCID mice . This immortalization required short-term P01133 treatment near the time of crisis . The permanent lines were P01133 -independent , but O75362 -dependent since siRNA to O75362 inhibited anchorage independence and arrested growth . Array CGH revealed genomic changes resembling those of ovarian carcinomas , such as amplicons at 3q and 20q , and deletions at 4q and 18 , associated with underexpressed annexin A10 , P19022 , desmocollin 3 and P05120 , which have been reported as tumor suppressors . The lines overexpressed Q05639 , SMARA3 and P42224 and underexpressed other oncogenes , tumor suppressors and extracellular matrix/adhesion genes . The results implicate O75362 as an ovarian oncogene , which is detrimental to senescing normal OSE cells but contributes to neoplastic progression in OSE with inactivated p53/RB . The resemblance of the genomic changes in the O75362 -overexpressing lines to ovarian carcinomas provides a unique model to investigate interrelationships between these changes and ovarian neoplastic phenotypes . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Identification of orally bioavailable , non-amidine inhibitors of DB00013 P00747 Activator ( uPA ) . In this Letter we report the synthesis and evaluation of a series of non-amidine inhibitors of DB00013 P00747 Activator ( uPA ) . Starting from compound 1 , a significant change provided compounds in which the amidine , binding in the S1 pocket , was replaced with a primary amine . Further modifications led to the identification of potent , selective , and orally bioavailable uPA inhibitors . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Proteomic identification of lynchpin urokinase plasminogen activator receptor protein interactions associated with epithelial cancer malignancy . DB00013 plasminogen activator ( uPA ) and its high affinity receptor ( Q03405 ) play crucial proteolytic and non-proteolytic roles in cancer metastasis . In addition to promoting plasmin-mediated degradation of extracellular matrix barriers , cell surface engagement of uPA through Q03405 binding results in the activation of a suite of diverse cellular signal transduction pathways . Because Q03405 is bound to the plasma membrane through a glycosyl-phosphatidylinositol anchor , these signalling sequelae are thought to occur through the formation of multi-protein cell surface complexes involving Q03405 . To further characterize Q03405 -driven protein complexes , we co-immunoprecipitated Q03405 from the human ovarian cancer cell line , OVCA 429 , and employed sensitive proteomic methods to identify the Q03405 -associated proteins . Using this strategy , we identified several known , as well as numerous novel , Q03405 associating proteins , including the epithelial restricted integrin , alphavbeta6 . Reverse immunoprecipitation using anti-beta6 integrin subunit monoclonal antibodies confirmed the co-purification of this protein with Q03405 . Inhibition of Q03405 and/or beta6 integrin subunit using neutralizing antibodies resulted in the inhibition of uPA-mediated P29323 1/2 phosphorylation and subsequent cell proliferation . These data suggest that the association of beta6 integrin ( and possibly other lynchpin cancer regulatory proteins ) with Q03405 may be crucial in co-transmitting uPA signals that induce cell proliferation . Our findings support the notion that Q03405 behaves as a lynchpin in promoting tumorigenesis by forming functionally active multiprotein complexes . P00749 is activated in stratum corneum after barrier disruption . The plasminogen/plasmin system in epidermis is thought to be the major protease involved in the delay of barrier recovery . However , little is known about the mechanism through which this system is activated . In order to clarify this mechanism , we first determined the distribution of proteolytic activity by using in situ zymography . As a result , plasminogen-activator activity was found to be present in the stratum corneum ( SC ) after barrier disruption . Next , SC subjected to repeated barrier disruption was collected to identify the protease . The protease was identified as urokinase-type plasminogen activator , because flybrinolytic activity of the collected SC was abolished by addition of anti-urokinase antibody . DB00013 activation in SC was confirmed by means of an in vitro assay , in which the precursor of urokinase ( pro-uPA ) became active after incubation with the insoluble component of SC homogenate . These findings indicated that urokinase-type plasminogen activator is activated in SC after barrier disruption and this activation might trigger the plasminogen/plasmin system in the epidermis . Silencing urokinase in the ventral tegmental area in vivo induces changes in cocaine-induced hyperlocomotion . DB00133 proteases in the nervous system have functional roles in neural plasticity . Among them , urokinase-type plasminogen activator ( uPA ) exerts a variety of functions during development , and is involved in learning and memory . Furthermore , psychostimulants strongly induce uPA expression in the mesolimbic dopaminergic pathway . In this study , doxycycline-regulatable lentiviruses expressing either uPA , a dominant-negative form of uPA , or non-regulatable lentiviruses expressing small interfering RNAs ( siRNAs ) targeted against uPA have been prepared and injected into the ventral tegmental area ( VTA ) of rat brains . Over-expression of uPA in the VTA induces doxycycline-dependent expression of its receptor , Q03405 , but not its inhibitor , plasminogen activator inhibitor-1 ( P05121 ) . Q03405 expression in the VTA is repressed upon silencing of uPA with lentiviruses expressing siRNAs . In addition , over-expression of uPA in the VTA promotes a 15-fold increase in locomotion activity upon cocaine delivery . Animals expressing the dominant-negative form of uPA did not display such hyperlocomotor activity . These cocaine-induced behavioural changes , associated with uPA expression , could be suppressed in the presence of doxycycline or uPA-specific siRNAs expressing lentiviruses . These data strongly support the major role of urokinase in cocaine-mediated plasticity changes . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . DB00013 and tissue plasminogen activators and their P05121 inhibitor in tumors of patients with oral mucosal cancer : relationship with the main clinical morphological factors . Enhanced activation of plasminogen by the urokinase pathway ( uPA elevation ) in patients with cancer of the oral mucosa paralleled by an increase of P05121 level in the tumor compared to the adjacent mucosa was shown by enzyme immunoassay . No statistically significant associations of the level of the studied proteins in the tumor with such prognostic factors as location , growth form , histological structure , differentiation degree , size , and dissemination of the primary tumor , involvement of the regional lymph nodes , and stage of the disease were detected . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . P02786 and P60709 as the best reference genes to quantify DB00013 P00747 Activator in breast cancer . BACKGROUND : Biomedical researchers have long looked for ways to diagnose and treat cancer patients at the early stages through biomarkers . Although conventional techniques are routinely applied in the detection of biomarkers , attitudes towards using Real-Time PCR techniques in detection of many biomarkers are increasing . Normalization of quantitative Real-Time PCR is necessary to validate non-biological alteration occurring during the steps of RNA quantification . Selection of variably expressed housekeeping genes ( HKs ) will affect the validity of the data . The aim of the present study was to identify uniformly expressed housekeeping genes in order to use in the breast cancer gene expression studies . DB00013 P00747 Activator was used as a gene of interest . FINDINGS : The expression of six HKs ( P02786 , P08236 , P04406 , P60709 , P00492 and P05388 ) was investigated using geNorm and NormFinder softwares in forty breast tumor , four normal and eight adjacent tissues . P05388 and P04406 revealed maximum M value , while P02786 demonstrated lowest M value . CONCLUSIONS : In the present study the most and the least stable genes were P02786 and P05388 respectively . P02786 and P60709 were verified as the best combination of two genes for breast cancer quantification . The result of this study shows that in each gene expression analysis HKs selection should be done based on experiment conditions . DB00013 receptor mediates doxorubicin-induced vascular smooth muscle cell senescence via proteasomal degradation of TRF2 . The anthracycline doxorubicin is a widely used effective anti-cancer drug . However , its application and dosage are severely limited due to its cardiotoxicity . The exact mechanisms of doxorubicin-induced cardiotoxic side effects remain poorly understood . Even less is known about the impact of doxorubicin treatment on vascular damage . We found that low doses of doxorubicin induced a senescent response in human primary vascular smooth muscle cells ( VSMC ) . We observed that expression of urokinase receptor ( Q03405 ) was upregulated in response to doxorubicin . Furthermore , the level of Q03405 expression played a decisive role in developing doxorubicin-induced senescence . Q03405 silencing in human VSMC by means of RNA interference as well as Q03405 knockout in mouse VSMC resulted in abrogation of doxorubicin-induced cellular senescence . On the contrary , Q03405 overexpression promoted VSMC senescence . We further found that proteasomal degradation of telomeric repeat binding factor 2 ( TRF2 ) mediates doxorubicin-induced VSMC senescence . Our results demonstrate that Q03405 controls the ubiquitin-proteasome system in VSMC and regulates doxorubicin-induced TRF2 ubiquitination and proteasomal degradation via this mechanism . Therefore , VSMC senescence induced by low doses of doxorubicin may contribute to vascular damage upon doxorubicin treatment . Q03405 -mediated TRF2 ubiquitination and proteasomal degradation are further identified as a molecular mechanism underlying this process . Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 agonist bromocriptine or a placebo in two separate sessions . DB01200 modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation . DB00013 plasminogen activator receptor is upregulated by Helicobacter pylori in human gastric cancer AGS cells via P29323 , JNK , and AP-1 . The gastric pathogen Helicobacter pylori ( H. pylori ) is suggested to be associated with gastric cancer progression . In this study , we investigated the effect of H. pylori on urokinase plasminogen activator receptor ( Q03405 ) expression which has been known to correlate closely with gastric cancer invasion . H. pylori induced the Q03405 expression in a time- and concentration-dependent manner . Specific inhibitors and inactive mutants of MEK-1 and JNK were found to suppress the H. pylori-induced Q03405 expression and the Q03405 promoter activity . Electrophoretic mobility shift assay and transient transfection study using an AP-1 decoy oligonucleotide confirmed that the activation of AP-1 is involved in the H. pylori-induced Q03405 upregulation . The AGS cells treated with H. pylori showed a remarkably enhanced invasiveness , and this effect was partially abrogated by Q03405 -neutralizing antibodies . These results suggest that H. pylori induces Q03405 expression via Erk-1/2 , JNK , and AP-1 signaling pathways and , in turn , stimulates the cell invasiveness in human gastric cancer AGS cells . DB00013 -dependent human vascular smooth muscle cell adhesion requires selective vitronectin phosphorylation by ectoprotein kinase CK2 . DB00013 ( uPA ) - and urokinase receptor ( Q03405 ) -dependent cell adhesion to the extracellular matrix protein vitronectin ( Vn ) is an important event in wound healing , tissue remodeling , immune response , and cancer . We recently demonstrated that in human vascular smooth muscle cells ( VSMC ) uPA/ Q03405 are functionally associated with the ectoprotein kinase casein kinase-2 ( CK2 ) . We now asked whether CK2 regulates uPA-dependent cell adhesion to Vn , since the latter is a natural CK2 substrate . We found that Vn is indeed selectively phosphorylated by CK2 and that this phosphorylation is uPA-regulated in VSMC . Vn induces release of ecto-CK2 from the cell surface via a process termed as " shedding. " CK2-mediated Vn phosphorylation was decisive for the uPA-dependent VSMC adhesion . Specific inhibition of CK2 completely abolished the uPA-induced cell adhesion to Vn . This effect was specific for cell adhesion to Vn and required participation of both Q03405 and alpha(v)beta(3) integrins as adhesion receptors . CK2 localization at the cell surface was highly dynamic ; Vn induced formation of clusters where CK2 colocalized with Q03405 and alpha(v)beta(3) integrins . These results indicate that the uPA-dependent VSMC adhesion is a function of selective Vn phosphorylation by the ectoprotein kinase CK2 and suggest a regulatory role for Vn phosphorylation in the uPA-directed adhesive process . Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . Dependence on phosphoinositide 3-kinase and DB01367 -RAF pathways drive the activity of RAF265 , a novel RAF/ P35968 inhibitor , and RAD001 ( DB01590 ) in combination . Activation of phosphatidylinositol-3-kinase ( PI3K ) -AKT and Kirsten rat sarcoma viral oncogene homologue ( P01116 ) can induce cellular immortalization , proliferation , and resistance to anticancer therapeutics such as epidermal growth factor receptor inhibitors or chemotherapy . This study assessed the consequences of inhibiting these two pathways in tumor cells with activation of P01116 , PI3K-AKT , or both . We investigated whether the combination of a novel RAF/vascular endothelial growth factor receptor inhibitor , RAF265 , with a mammalian target of rapamycin ( P42345 ) inhibitor , RAD001 ( everolimus ) , could lead to enhanced antitumoral effects in vitro and in vivo . To address this question , we used cell lines with different status regarding P01116 , P42336 , and P15056 mutations , using immunoblotting to evaluate the inhibitors , and MTT and clonogenic assays for effects on cell viability and proliferation . Subcutaneous xenografts were used to assess the activity of the combination in vivo . RAD001 inhibited P42345 downstream signaling in all cell lines , whereas RAF265 inhibited RAF downstream signaling only in P15056 mutant cells . In vitro , addition of RAF265 to RAD001 led to decreased AKT , S6 , and P06730 binding protein 1 phosphorylation in HCT116 cells . In vitro and in vivo , RAD001 addition enhanced the antitumoral effect of RAF265 in HCT116 and H460 cells ( both P01116 mut , P42336 mut ) ; in contrast , the combination of RAF265 and RAD001 yielded no additional activity in A549 and MDAMB231 cells . The combination of RAF and P42345 inhibitors is effective for enhancing antitumoral effects in cells with deregulation of both DB01367 -RAF and PI3K , possibly through the cross-inhibition of 4E binding protein 1 and S6 protein . DB00013 binds to a plasminogen activator inhibitor type-2-like molecule in placental microvillous membranes . Placental microvillous membranes exhibited saturable binding of urokinase-type plasminogen activator with plateau achieved by 30 min at 4 degrees C and 10 min at 37 degrees C . The binding was essentially irreversible . The capacity was about 8 pmol urokinase per mg membrane protein . Half-maximal displacement of 125I-labelled urokinase was achieved with about 1.0 nM unlabelled urokinase when using 75 micrograms membrane protein/ml . 125I-labelled urokinase did not bind when treated with diisopropylfluorophosphate to block the catalytic activity . Single-chain urokinase ( prourokinase ) , devoid of catalytic activity , did not bind . Catalytically active tissue-type plasminogen activator did compete with 125I-labelled urokinase for binding although less efficiently than urokinase . Binding activity remained in the 100,000 x g pellet after treatment of the membranes with 3 M DB00761 , alkaline stripping at pH 12 or extraction by the detergent Triton X-100 . The binding was essentially blocked by antibodies against plasminogen activator inhibitor-type-2 ( P05120 ) . DB00815 polyacrylamide gel electrophoresis of solubilized membranes with bound 125I-labelled urokinase showed that the urokinase- P05120 complexes largely migrated in fractions corresponding to a very large Mr although no clearly defined peaks were observed . It is suggested that P05120 occurs in a form anchored to syncytiotrophoblast microvilli , possibly to the cytoskeleton . P00747 activator production in a rat model of Pneumocystis carinii pneumonia . Several studies have indicated that the serine protease urokinase-plasminogen-activator ( uPA ) is an important factor in host defense against pulmonary pathogens . To gain a better insight into the role of uPA in Pneumocystis carinii ( P. carinii ) pneumonia ( PCP ) , we evaluated PA production in alveolar macrophages ( AMs ) obtained from rats with steroid-induced PCP . Treatment with cortisone acetate favored PCP in 91 % of rats . In the bronchoalveolar lavage ( BAL ) samples of immunosuppressed rats both with and without PCP , we observed a decrease in uPA activity as well as a decrease in cell number . DB00013 -PA production by AMs was reduced in rats treated with cortisone alone . However , an increase in cell-associated uPA was observed in rats with PCP . This increase appears to be produced in response to P carinii infection . In fact , when AMs obtained from untreated healthy or immunosuppressed uninfected rats were challenged with P carinii , a significant increase in PA activity in cell lysates was observed , though a lower response was obtained in cortisone-treated animals . Our results suggest that healthy AMs respond to the presence of P carinii with an increase in uPA production and that this response in immunodepressed rat-AMs is partially impaired . P00533 is a transducer of the urokinase receptor initiated signal that is required for in vivo growth of a human carcinoma . DB00013 plasminogen activator receptor ( Q03405 ) activates alpha5beta1 integrin and P29323 signaling , inducing in vivo proliferation of HEp3 human carcinoma . Here we demonstrate that P00533 mediates the Q03405 /integrin/fibronectin ( FN ) induced growth pathway . Its activation is ligand-independent and does not require high P00533 , but does require high Q03405 expression . Only when Q03405 level is constitutively elevated does P00533 become alpha5beta1-associated and activated . Domain 1 of Q03405 is crucial for P00533 activation , and Q05397 links integrin and P00533 signaling . Inhibition of P00533 kinase blocks Q03405 induced signal to P29323 , implicating P00533 as an important effector of the pathway . Disruption of Q03405 or P00533 signaling reduces HEp3 proliferation in vivo . These findings unveil a mechanism whereby Q03405 subverts ligand-regulated P00533 signaling , providing cancer cells with proliferative advantage . Interferon-γ promotes vascular remodeling in human microvascular endothelial cells by upregulating endothelin ( ET ) -1 and transforming growth factor ( TGF ) β2 . Systemic sclerosis ( SSc ) is a complex disease characterized by vascular alterations , activation of the immune system and tissue fibrosis . Previous studies have implicated activation of the interferon pathways in the pathogenesis of SSc . The goal of this study was to determine whether interferon type I and/or type II could play a pathogenic role in SSc vasculopathy . Human dermal microvascular endothelial cells ( HDMVECs ) and fibroblasts were obtained from foreskins of healthy newborns . The RT Profiler PCR Array System was utilized to screen for EndoMT genes . Treatment with IFN-α or IFN-γ downregulated Fli1 and P33151 . In contrast , IFN-α and IFN-γ exerted opposite effects on the expression of α-SMA , P29279 , ET-1 , and TGFβ2 , with IFN-α downregulating and IFN-γ upregulating this set of genes . Blockade of TGFβ signaling normalized IFN-γ-mediated changes in Fli1 , P33151 , P29279 , and ET-1 levels , whereas upregulation of α-SMA and TGFβ2 was not affected . DB00559 treatment was more effective than TGFβ blockade in reversing the actions of IFN-γ , including downregulation of α-SMA and TGFβ2 , suggesting that activation of the ET-1 pathway plays a main role in the IFN-γ responses in HDMECs . IFN-γ induced expression of selected genes related to endothelial-to-mesenchymal transition ( EndoMT ) , including Snail1 , P02751 , P05121 , Q15672 , P40763 , P41220 , and components of the WNT pathway . The effect of IFN-γ on EndoMT was mediated via TGFβ2 and ET-1 signaling pathways . This study demonstrates distinct effects of IFN-α and IFN-γ on the biology of vascular endothelial cells . IFN-γ may contribute to abnormal vascular remodeling and fibrogenesis in SSc , partially via induction of EndoMT . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . MiR-221/-222 differentiate prognostic groups in advanced breast cancers and influence cell invasion . BACKGROUND : MiR-221/-222 are frequently overexpressed in breast cancer and are associated with increased malignancy . The specific modification of microRNAs ( miRNAs ) expression could be a promising strategy in breast cancer therapy , leading to the suppression of tumourigenic processes in tumour cells . METHODS : MiR-221/-222 expressions were analysed in 86 breast cancer tissues by quantitative RT-PCR and tested for correlation with immunohistochemistry data and clinical follow-up . In vitro assays were conducted using human breast cancer cell lines with lentiviral overexpression of miR-221/-222 . RESULTS : In tumour tissues , miR-221/-222 were associated with the occurrence of distant metastases . In particular , high levels of miR-221 were revealed to have a high prognostic impact for the identification of significantly different groups with advanced tumours . MiR-221/-222 overexpression strongly increased cell proliferation and invasion in vitro . Following miR-221/-222 overexpression an increased Q03405 expression and cell invasion were observed . CONCLUSION : This study demonstrates a significant role for highly expressed miR-221/-222 in advanced breast cancers allowing for the identification of significantly different prognostic groups , particularly for P04626 -positive and lymph-node-positive breast cancers . Considering that miR-221/-222 are strongly involved in cell invasion , these miRNAs may be promising markers for breast cancer prognosis and therapy . Amyloid beta-derived neuroplasticity in bone marrow-derived DB05914 is mediated by P01303 and P41595 receptors via P27361 /2 signalling pathways . OBJECTIVE : In Alzheimer 's disease , toxic soluble and insoluble forms of amyloid beta ( Abeta ) cause synaptic dysfunction and neuronal loss . Given its potential role in producing a toxic host microenvironment for transplanted donor stem cells , we investigated the interaction between Abeta and proliferation , survival , and differentiation of bone marrow-derived DB05914 ( BM- O60682 ) in culture . MATERIALS AND METHODS : We used BM- O60682 that had been isolated from mouse bone marrow and cultured , and we also assessed relevant reaction mechanisms using gene microarray , immunocytochemistry , and inhibitors of potential signalling molecules , such as mitogen-activated protein kinase ( MAPK ) /extracellular signal-regulated kinase ( P29323 )1/2 and tyrosine protein kinase . RESULTS AND CONCLUSIONS : Interestingly , we found that treatment with aggregated ( 1-40 or 1-42 ) and oligomeric ( 1-42 ) Abeta promoted neuronal-like differentiation of BM- O60682 without toxic effects . This was not dependent on soluble factors released from BM- O60682 progeny nor solely on formation of Abeta fibrils . The effect of Abeta is mediated by G-protein coupled receptors , neuropeptide Q03519 ( P25929 ) and serotonin ( 5-hydroxytryptamine ) receptor 2B , via phosphatidylinositol-3-OH kinase-dependent activation of the MAPK/ P27361 /2 . Our results lend support to the idea that reciprocal donor stem cell-host interactions may promote a regenerative response that can be exploited by epigenetic modulation of P01303 /serotonergic gene expression , for stem cell therapy , in Alzheimer 's disease . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . Angiogenic alterations associated with circulating neoplastic DNA in ovarian carcinoma . OBJECTIVES : Forty percent of women with ovarian carcinoma have circulating free neoplastic DNA identified in plasma . Angiogenesis is critical in neoplastic growth and metastasis . We sought to determine whether circulating neoplastic DNA results from alterations in the balance of angiogenesis activators and inhibitors . METHODS : Sixty patients with invasive ovarian carcinomas with somatic P04637 mutations that had been characterized for circulating neoplastic DNA had carcinoma analyzed for microvessel density using immunohistochemistry with CD31 and for the expression of P15692 , Q15389 , O15123 , P35354 , P00749 , P07996 , P09603 , P42336 , Q16665 , P10145 , P08253 , and P14780 message by real-time quantitative polymerase chain reaction . The expression of each gene was calculated relative to P04406 expression for each neoplasm . Patient plasma had been tested for circulating neoplastic DNA using a ligase detection reaction . RESULTS : P08253 expression was significantly correlated with free plasma neoplastic DNA ( P = .007 ) . Microvessel density was not correlated with plasma neoplastic DNA or P38398 /2 mutation status . The expression pattern of other angiogenic factors did not correlate with plasma neoplastic DNA but correlated with each other . P38398 /2 mutated carcinomas had significantly different expression profiles of angiogenesis activators and inhibitors in comparison to sporadic carcinomas . CONCLUSIONS : P08253 expression is associated with the presence of circulating neoplastic DNA in women with ovarian carcinoma . These data are consistent with the proinvasive properties of P08253 and suggest that the presence of circulating neoplastic DNA indicates a more aggressive malignant phenotype . Carcinomas with germ line P38398 /2 mutations had a lower angiogenic profile than those without mutations . P00747 acceleration of urokinase thrombolysis . PURPOSE : A relative deficiency of plasminogen within the thrombus may be the rate limiting factor in clot lysis . METHODS : To investigate this hypothesis , we used an in vitro perfusion system and expanded polytetrafluoroethylene graft segments filled with radiolabeled human thrombus . Three groups of five perfusions were compared : ( 1 ) urokinase infusion ( 333 IU/min ) into clots laced with buffer , ( 2 ) urokinase infusion ( 333 IU/min ) into clots laced with plasminogen ( 44 CU ) , and ( 3 ) control , D5W infusion into clots laced with buffer . Two end points were measured over time : the amount of lysed thrombus and the flow through the graft . RESULTS : DB00013 infusion resulted in augmented flow through the graft when compared with control ( p < 0.05 ) . Lacing with plasminogen resulted in more rapid restoration of flow when compared with urokinase infusion alone ( p < 0.05 ) . Similarly , the rate of clot dissolution was significantly greater in plasminogen-laced thrombi ( p < 0.05 ) when compared with the control and urokinase groups . Embolization of particles of thrombus was uniformly observed in the urokinase group , resulting in a temporary decrease in flow through the thrombosed graft . This event characteristically occurred after 60 minutes of infusion but was never seen in the urokinase/plasminogen treatment group . CONCLUSIONS : These results suggest that plasminogen supplementation of urokinase thrombolysis may result in significant clinical benefits with respect to the rate of clot lysis and the uniformity of clot dissolution with a lower likelihood of secondary embolization . MEK/ P29323 -dependent Q03405 expression is required for motility via phosphorylation of P70S6K in human hepatocarcinoma cells . Motility and invasiveness events require specific intracellular signaling cascade activations . In cancer liver cells , one of these mechanisms could involve the MAPK MEK/ P29323 cascade activation which has been shown over expressed and activated in hepatocellular carcinoma . To study whether the MEK/ P29323 cascade is involved in the motility of HCC , we examined the effect of MEK inhibitor and P28482 silencing using monolayer wound-healing assays and fluoroblock invasion systems . Evidence was provided that the MAPK cascade is a key transduction pathway which controls HCC cells motility and invasiveness . We could disconnect proliferation to motility using mitomycin C and we established that RNAi-mediated inhibition of P28482 led to strongly reduced cell motility . To improve our understanding , we analysed the regulation and the role of urokinase receptor ( Q03405 ) in this process . We provided evidence that Q03405 was under a MEK/ P29323 dependent mechanism and blocking Q03405 activity using specific antagonist or inhibiting its expression by RNA interference which resulted in complete inhibition of motility . Moreover , we found in MAPK inhibited cultures and in Q03405 silencing cells that p70S6K phosphorylation on residue DB00156 -389 was significantly reduced , whereas DB00133 -421/ DB00156 -424 phosphorylation did not change . We highlighted that the P42345 / P42345 pathway did not affect motility and DB00156 -389 phosphorylation . Furthermore , we demonstrated that p70S6K inhibition by RNA interference completely inhibited hepatocarcinoma cell motility . Therefore , targeting Q03405 and/or MEK/ P29323 /S6K by RNA interference could be a major therapeutic strategy for the future treatment of invasive hepatocarcinoma cells . DB00013 receptor splice variant Q03405 -del4/5-associated gene expression in breast cancer : identification of rab31 as an independent prognostic factor . PURPOSE : To evaluate the pure prognostic impact of the uPA-receptor splice variant Q03405 -del4/5 for lymph node-negative breast cancer patients , and to identify differentially expressed genes associated with high or low Q03405 -del4/5 mRNA levels . PATIENTS AND METHODS : mRNA transcript levels were measured by real-time PCR in tumor samples from 280 node-negative breast cancer patients who had not received adjuvant systemic therapy . Endpoints were distant metastasis-free survival ( DMFS ) and overall survival ( OS ) . Gene expression analysis was performed with RNA isolated from breast cancer tissue and breast cancer cell lines using Affymetrix U133a GeneChips . RESULTS : In multivariate analysis , Q03405 -del4/5 significantly contributed to the base model of traditional prognostic factors for DMFS ( HR = 3.29 , P < 0.001 ) and OS ( HR = 2.87 , P = 0.002 ) . Using microarrays , seven genes were found to be up-regulated in tumor samples and cancer cell lines with high Q03405 -del4/5 mRNA expression . The gene encoding rab31 , a member of the Ras oncogene family , was selected for quantitative analysis of mRNA expression in the set of 280 patients . High rab31 values were significantly associated with worse outcome of patients for DMFS ( HR = 2.27 , P < 0.001 ) and OS ( HR = 2.01 , P = 0.008 ) in multivariate analysis , independent from Q03405 -del4/5 . The patient subgroup with high Q03405 -del4/5 and rab31 levels showed the worst DMFS and OS ( P < 0.001 , both ) compared with tumors with low values of both factors . CONCLUSIONS : Our results suggest that Q03405 -del4/5 and rab31 mRNA represent independent prognostic markers in breast cancer and may be components of different , but possibly associated , tumor-relevant signaling pathways . DB00013 receptor ( CD87 ) regulates leukocyte recruitment via beta 2 integrins in vivo . The urokinase receptor ( CD87 ; Q03405 ) is found in close association with beta 2 integrins on leukocytes . We studied the functional consequence of this association for leukocyte adhesion and migration . In vivo , the beta 2 integrin-dependent recruitment of leukocytes to the inflamed peritoneum of Q03405 -deficient mice was significantly reduced as compared with wild-type animals . In vitro , beta 2 integrin-mediated adhesion of leukocytes to endothelium was lost upon removal of Q03405 from the leukocyte surface by phosphatidyl-inositol-specific phospholipase C . Leukocyte adhesion was reconstituted when soluble intact Q03405 , but not a truncated form lacking the uPA-binding domain , was allowed to reassociate with the cell surface . Q03405 ligation with a monoclonal antibody induced adhesion of monocytic cells and neutrophils to vascular endothelium by six- to eightfold , whereas ligation with inactivated uPA significantly reduced cell-to-cell adhesion irrespective of the beta 2 integrin-stimulating pathway . These data indicate that beta 2 integrin-mediated leukocyte-endothelial cell interactions and recruitment to inflamed areas require the presence of Q03405 and define a new phenotype for Q03405 -deficient mice . Moreover , Q03405 ligation differentially modulates leukocyte adhesion to endothelium and provides novel targets for therapeutic strategies in inflammation-related vascular pathologies . P00749 ( uPA ) induces pulmonary microvascular endothelial permeability through low density lipoprotein receptor-related protein ( Q14764 ) -dependent activation of endothelial nitric-oxide synthase . DB00013 plasminogen activator ( uPA ) and PA inhibitor type 1 ( P05121 ) are elevated in acute lung injury , which is characterized by a loss of endothelial barrier function and the development of pulmonary edema . Two-chain uPA and uPA- P05121 complexes ( 1-20 nM ) increased the permeability of monolayers of human pulmonary microvascular endothelial cells ( PMVECs ) in vitro and lung permeability in vivo . The effects of uPA- P05121 were abrogated by the nitric-oxide synthase ( NOS ) inhibitor L-NAME ( N(D)-nitro-L-arginine methyl ester ) . Two-chain uPA ( 1-20 nM ) and uPA- P05121 induced phosphorylation of endothelial NOS- DB00133 (1177) in PMVECs , which was followed by generation of NO and the nitrosylation and dissociation of β-catenin from P33151 . uPA-induced phosphorylation of P29474 was decreased by anti-low density lipoprotein receptor-related protein-1 ( Q14764 ) antibody and an Q14764 antagonist , receptor-associated protein ( P30533 ) , and when binding to the uPA receptor was blocked by the isolated growth factor-like domain of uPA. uPA-induced phosphorylation of P29474 was also inhibited by the protein kinase A ( PKA ) inhibitor , myristoylated PKI , but was not dependent on PI3K-Akt signaling . Q14764 blockade and inhibition of PKA prevented uPA- and uPA- P05121 -induced permeability of PMVEC monolayers in vitro and uPA-induced lung permeability in vivo . These studies identify a novel pathway involved in regulating PMVEC permeability and suggest the utility of uPA-based approaches that attenuate untoward permeability following acute lung injury while preserving its salutary effects on fibrinolysis and airway remodeling .

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interacts_with DB00054?

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The high molecular mass , glycoprotein Ib-binding protein flavocetin-A induces only small platelet aggregates in vitro . The direct effects of snake venom glycoprotein ( GP ) Ib-binding proteins on platelet receptors during the formation of platelet aggregates were determined by a particle counting method using light scattering . Flavocetin-A induces small platelet aggregates , but not medium or large ones . However , neither jararaca GPIb-BP nor tokaracetin induce platelet aggregation . The flavocetin-A dose-response curve for formation of small aggregates is bell-shaped , with maximal effect at 1 to 2 microg/mL . The formation of small aggregates was not observed when fixed human platelets were used . Jararaca GPIb-BP , the anti-GPIb monoclonal antibody GUR83-35 , prostaglandin I2 , and ethylene diamine-N,N-dimethylformamide all inhibited flavocetin-A-induced small aggregate formation , but acetylsalicylic acid did not . Furthermore , anti- P08514 /IIIa monoclonal antibodies , DB00054 , and YM337 significantly but partially inhibited aggregate formation , but the anti- P04275 monoclonal antibody NMC-4 had no effect . The formation of small aggregates required extracellular calcium , but flavocetin-A did not elevate cytosolic calcium . These results suggest that flavocetin-A binds to intact platelets , initiating platelet responses and inducing platelet aggregate formation by cross-linking platelets . Consequently , flavocetin-A may be a useful tool to study the mechanism of GPIb-mediated platelet activation and the structure-function relationships of GPIb . Comparative studies of a humanized anti-glycoprotein IIb/IIIa monoclonal antibody , YM337 , and abciximab on in vitro antiplatelet effect and binding properties . The effects of YM337 , the Fab fragment of a humanized anti-glycoprotein IIb/IIIa ( P08514 /IIIa ) monoclonal antibody C4G1 , on in vitro platelet function and binding properties were compared with those of abciximab , the Fab fragment of the human/murine chimeric anti- P08514 /IIIa monoclonal antibody DB00054 . Both agents completely inhibited platelet aggregation caused by all agonists tested except ristocetin . Further , both inhibited human platelet adhesion to P04275 , fibrinogen , fibronectin and subendothelial matrix with similar potency . DB09222 binding to washed platelets was dose-dependently inhibited by both agents . In binding assay using 125I-YM337 and 125I-abciximab , Kd values determined with platelet-rich plasma were 6.74 +/- 0.56 nM for YM337 and 6.65 +/- 1.45 nM for abciximab , and the number of binding sites were 42,700 +/- 3,000 for YM337 and 76,000 +/- 5,400 for abciximab . P08514 /IIIa was precipitated from the solubilized fraction of platelets by both agents . In contrast , integrin alphavbeta3 was precipitated from the solubilized fraction of human umbilical vein endothelial cells by abciximab but not by YM337 . DB09222 binding to purified P08514 /IIIa was dose-dependently inhibited by both agents . In contrast , vitronectin binding to purified integrin alphavbeta3 was dose-dependently inhibited by abciximab but not by YM337 , supporting the idea that abciximab reacts to integrin alphavbeta3 . Therefore , YM337 was suggested to bind to a different epitope of P08514 /IIIa from abciximab . These results suggest that YM337 specifically acts on platelet P08514 /IIIa receptors and has similar inhibitory properties on platelet aggregation and platelet adhesion to abciximab . Increased thromboxane biosynthesis during coronary thrombolysis . Evidence that platelet activation and thromboxane A2 modulate the response to tissue-type plasminogen activator in vivo . Platelet activation is markedly increased during coronary thrombolysis and limits the response to thrombolytic therapy . A possible mediator of platelet activation in this setting is thromboxane ( TX ) A2 , a potent platelet agonist formed in greatly increased amounts during coronary thrombolysis in man . To address this hypothesis , we examined the role of TXA2 in modulating the response to intravenous tissue-type plasminogen activator ( t-PA ) in a chronic canine model of coronary thrombosis . Reperfusion occurred in 60 +/- 5 minutes and was complicated by spontaneous reocclusion . The times to reperfusion and reocclusion were platelet-dependent . Consistent with a role for TXA2 in this process , TXA2 biosynthesis , determined a excretion of its enzymatic metabolite , 2,3-dinor-TXB2 , was markedly increased during coronary thrombolysis . Furthermore , inhibition of TXA2 by aspirin , given alone or in combination with a TXA2/prostaglandin endoperoxide receptor antagonist , accelerated reperfusion and partly inhibited cyclic flow variations during reperfusion . The delay in reperfusion and reocclusion induced by TXA2 appeared to be mediated by platelet aggregation since the F(ab')2 fragment of DB00054 , a monoclonal antibody to the platelet P08514 /IIIa , also accelerated reperfusion and prevented reocclusion without altering TXA2 biosynthesis . These finding suggest that platelet aggregation limits the response to coronary thrombolysis and that platelet activation in this setting is partly TXA2-dependent . P00533 variant III mutations in lung tumorigenesis and sensitivity to tyrosine kinase inhibitors . The tyrosine kinase inhibitors gefitinib ( DB00317 ) and erlotinib ( Tarceva ) have shown anti-tumor activity in the treatment of non-small cell lung cancer ( NSCLC ) . Dramatic and durable responses have occurred in NSCLC tumors with mutations in the tyrosine kinase domain of the epidermal growth factor receptor ( P00533 ) . In contrast , these inhibitors have shown limited efficacy in glioblastoma , where a distinct P00533 mutation , the variant III ( vIII ) in-frame deletion of exons 2-7 , is commonly found . In this study , we determined that EGFRvIII mutation was present in 5 % ( 3/56 ) of analyzed human lung squamous cell carcinoma ( SCC ) but was not present in human lung adenocarcinoma ( 0/123 ) . We analyzed the role of the EGFRvIII mutation in lung tumorigenesis and its response to tyrosine kinase inhibition . Tissue-specific expression of EGFRvIII in the murine lung led to the development of NSCLC . Most importantly , these lung tumors depend on EGFRvIII expression for maintenance . Treatment with an irreversible P00533 inhibitor , HKI-272 , dramatically reduced the size of these EGFRvIII-driven murine tumors in 1 week . Similarly , Ba/ P13726 cells transformed with the EGFRvIII mutant were relatively resistant to gefitinib and erlotinib in vitro but proved sensitive to HKI-272 . These findings suggest a therapeutic strategy for cancers harboring the EGFRvIII mutation . Repetitive profound thrombocytopenia after treatment with tirofiban : a case report . The P08514 /IIIa inhibitors are used in the acute coronary syndromes and interventional cardiology as antiplatelet agents . These drugs induce thrombocytopenia in approximately 1-5 % of patients . Thrombocytopenia is rapid in onset and antibody mediated . DB00054 is associated with higher incidence of thrombocytopenia than eptifibatide and tirofiban . Profound thrombocytopenia has reportedly been an issue with abciximab , but not with tirofiban . We reported a case of acute profound thrombocytopenia due to on tirofiban treatment in the same patient at two different times . Antiplatelet activity of nifedipine is mediated by inhibition of NF-κB activation caused by enhancement of Q07869 -β/-γ activity . BACKGROUND AND PURPOSE : The transcription factor NF-κB , stimulates platelet aggregation through a non-genomic mechanism . Nifedipine , a voltage-gated L-type calcium channel blocker , is widely used to treat hypertension . Nifedipine also displays antiplatelet activity , but the underlying mechanisms involved remain unclear . This study was designed to investigate whether the antiplatelet effects of nifedipine are mediated by regulating NF-κB-dependent responses . EXPERIMENTAL APPROACH : Platelet aggregation was measured turbidimetrically using an aggregometer . NF-κB and Q07869 activation , intracellular Ca2+ mobilization , PKCα activity , surface glycoprotein IIb/IIIa ( P08514 /IIIa ) expression and platelet activation-related signalling pathways were determined in control and nifedipine-treated platelets in the presence or absence of Q07869 antagonists or betulinic acid , a NF-κB activator . KEY RESULTS : Exposure of platelets to nifedipine significantly increased the Q07869 -β/-γ activity in activated human platelets . Treatment with nifedipine reduced collagen-induced NF-κB events , including the phosphorylation of IκB kinase-β , IκBα and p65NF-κB , which were markedly attenuated by GSK0660 , a Q07869 -β antagonist , or GW9662 , a Q07869 -γ antagonist . Furthermore , the interaction of Q07869 -β/-γ with NF-κB and the Q07869 -β/-γ-up-regulated NO/cGMP/PKG1 cascade may contribute to inhibition of NF-κB activation by nifedipine . Suppressing Q07869 -β/-γ activity or increasing NF-κB activation greatly reversed the inhibitory effect of nifedipine on collagen-induced platelet aggregation , intracellular Ca2+ mobilization , PKCα activity and surface P08514 /IIIa expression.CONCLUSIONS AND IMPLICATIONSPPAR-β/-γ-dependent inhibition of NF-κB activation contributes to the antiplatelet activity of nifedipine . These findings provide a novel mechanism underlying the beneficial effects of nifedipine on platelet hyperactivity-related vascular and inflammatory diseases . The effects of P08514 -IIIa antagonists and a combination of three other antiplatelet agents on platelet-leukocyte interactions . The effects of the P08514 -IIIa antagonists abciximab and MK-852 on platelet-leukocyte interactions in vitro were studied and the results compared with those obtained with a combination of aspirin , dipyridamole and AR-C69931 ( DB00128 /Dip/AR-C ) . Platelet-monocyte ( P/M ) and platelet-neutrophil ( P/N ) conjugate formation increased when blood was stirred or a platelet agonist was added . Leukocyte activation also occurred as judged by expression of surface tissue factor antigen and CD11b . DB00054 and MK-852 potentiated P/M , especially when collagen was used . They also increased the amount of tissue factor on the monocytes , but not CD11b . The DB00128 /Dip/AR-C did not enhance P/M or tissue factor exposure . Augmented tissue factor expression on monocytes in the presence of a P08514 -IIIa antagonist may be relevant to the increased mortality associated with trials of such antagonists when given orally in patients with vascular disease . The DB00128 /Dip/AR-C was superior to abciximab and MK-852 in inhibiting platelet and leukocyte function . Neonatal platelets are less reactive than adult platelets to physiological agonists in whole blood . Previous studies have reported that the platelets of healthy term neonates have either diminished or normal reactivity compared to the platelets of adults . To circumvent the methodologic problems of previous studies , we used a whole blood flow cytometric method to study neonatal platelet reactivity to thrombin , a combination of ADP and epinephrine , and U46619 ( a stable thromboxane A2 analogue ) . Inclusion in the assay of the peptide GPRP ( an inhibitor of fibrin polymerization ) enabled us to study platelet reactivity to human alpha-thrombin in whole blood . Umbilical cord blood and day 1 peripheral blood were collected from 30 healthy term neonates and compared to peripheral blood from 20 normal adults . In whole blood samples without added agonist , there were no significant differences between neonates and adults in the platelet binding of monoclonal antibodies 6D1 ( GPIb-specific ) or DB00054 ( P08514 -IIIa complex-specific ) . As determined by P28222 ( a P16109 -specific monoclonal antibody ) , neither neonates nor adults had circulating degranulated platelets . However , in both cord and peripheral whole blood samples , neonatal platelets were significantly less reactive than adult platelets to thrombin , ADP/epinephrine , and U46619 , as determined by the extent of increase in the platelet surface expression of P16109 and the P08514 -IIIa complex , and the extent of decrease in the platelet surface expression of the GPIb-IX complex. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00054 , eptifibatide , and tirofiban exhibit dose-dependent potencies to dissolve platelet aggregates . Platelet P08514 /IIIa antagonists are not only used to prevent platelet aggregation , but also in combination with thrombolytic agents for the treatment of coronary thrombi . Recent data indicate a potential of abciximab alone to dissolve thrombi in vivo . We investigated the potential of abciximab , eptifibatide , and tirofiban to dissolve platelet aggregates in vitro . DB00640 diphosphate ( ADP ) -induced platelet aggregation could be reversed in a concentration-dependent manner by all three P08514 /IIIa antagonists when added after the aggregation curve reached half-maximal aggregation . The concentrations chosen are comparable with in vivo plasma concentrations in clinical applications . Disaggregation reached a maximum degree of 72.4 % using 0.5 microg/ml tirofiban , 91.5 % using 3.75 microg/ml eptifibatide , and 48.4 % using 50 microg/ml abciximab ( P < 0.05 , respectively ) . A potential fibrinolytic activity of the P08514 /IIIa antagonists was ruled out by preincubation with aprotinin or by a plasma clot assay . A stable model Chinese hamster ovary ( CHO ) cell line expressing the activated form of P08514 /IIIa was used to confirm the disaggregation capacity of P08514 /IIIa antagonists found in platelets . Not only abciximab , but also eptifibatide and tirofiban have the potential to disaggregate newly formed platelet clusters in vitro . Because enzyme-dependent fibrinolysis does not appear to be involved , competitive removal of fibrinogen by the receptor antagonists is the most likely mechanism . Prevention of rethrombosis after coronary thrombolysis in a chronic canine model . I . Adjunctive therapy with monoclonal antibody 7E3 F(ab')2 fragment . We examined the efficacy of the monoclonal antibody ( MoAb ) 7E3 F(ab')2 fragment , an inhibitor of the platelet glycoprotein (GP)IIb/IIIa receptor , to prevent coronary artery rethrombosis after successful thrombolysis with rt-PA . The circumflex coronary artery of anesthetized dogs was instrumented with a flow probe , an electrode , and a stenosis . After recovery from the surgical procedure , the animals were reanesthetized on post-operative day 9 , and vessel wall injury was induced with current applied to the intimal surface of the circumflex coronary artery . The resulting occlusive thrombus was aged for 30 min , and recombinant tissue plasminogen activator ( rt-PA ) was administered . The animals were allocated to receive either placebo or a single dose of DB00054 [ 0.8 mg/kg intravenous ( i.v. ) bolus ] as the sole adjunctive agent . Ex vivo platelet function and coronary artery blood flow velocity were recorded on each of 5 consecutive days . Reocclusion and mortality were reduced significantly in animals treated with 7E3 as compared with the placebo-treated group . Significant inhibition of ex vivo platelet aggregation persisted for 48 h after a single injection of DB00054 . The MoAb 7E3 F(ab')2 fragment is effective as the sole adjunctive agent with rt-PA for prevention of rethrombosis . The present study is unique in that it examined the efficacy of P08514 /IIIa inhibition in an experimental model for an extended time , demonstrating the duration of antiplatelet therapy required to prevent rethrombosis after thrombolysis . DB00054 : a reappraisal of its use in coronary care . Platelet reactivity plays a pivotal role in the pathogenesis of ischemic adverse events during and after acute coronary syndromes ( ACS ) , and percutaneous coronary intervention ( P05154 ) . Glycoprotein ( GP ) IIb/IIIa inhibitors are the strongest antiplatelet agents currently available on the market and three different compounds , namely abciximab , tirofiban , and eptifibatide , have been approved for clinical use . DB00054 has been investigated in the clinical field far more extensively than the other P08514 /IIIa inhibitors . DB00054 is an anti-integrin Fab fragment of a human - mouse chimeric monoclonal antibody with high affinity and a slow dissociation rate from the GP IIb/IIIa platelet receptor . DB00054 , given shortly before the coronary intervention , is superior to placebo in reducing the acute risk of ischemic complications ( EPIC , EPISTENT , EPILOG trials ) ; moreover , in the ISAR-REACT 2 study abciximab has been shown to reduce the risk of adverse events in patients with non ST-segment elevation ACS who are undergoing P05154 even after optimal pre-treatment with 600 mg of clopidogrel . Finally , abciximab has been also used in abciximab-coated stent , with only bolus administration regimen and for direct intracoronary use with promising results that may extend and/or modify its current use in clinical practice in future . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Delayed immunologic thrombocytopenia induced by abciximab . DB00054 is an anti- P08514 -IIIa drug widely used to prevent thrombotic complications during percutaneous coronary intervention . We now report on the immunologic origin of thrombocytopenia developing between 7 and 12 days after the onset of abciximab infusion . Antibodies directed against abciximabcoated platelets were located in 5 patients with delayed thrombocytopenia , just as they were present in a patient whose platelet count fell within a few hours after receiving the drug . DB00054 -dependent IgG antibody was revealed in serum using control platelets in the monoclonal antibody immobilization of platelet antigens assay ( MAIPA ) performed with SZ22 , a MoAb to P08514 . The presence of IgG antibodies specific for platelets sensitized with abciximab was confirmed by flow cytometry . They were not located in 13 patients receiving abciximab but whose platelet counts remained stable . For three patients , antibodies were transient and their presence related to the extent of the thrombocytopenia . Surprisingly , antibodycontaining plasma from three patients induced abciximabdependent activation and aggregation of normal platelets , a finding confirmed by electron microscopy . Immunogold labeling revealed that abciximab was associated with platelets in the aggregate , suggesting that its inhibitory effect was overcome by the platelet stimulation . In summary , these results show that abciximab-dependent thrombocytopenia can be delayed and potentially prothrombotic . The first two cases of neonatal alloimmune thrombocytopenia associated with the low-frequency platelet antigen Q9Y251 -21bw ( Nos ) in Japan . BACKGROUND : Neonatal alloimmune thrombocytopenia ( NAIT ) is a disorder characterized by maternal alloimmunization against paternal fetal platelet antigens . Two healthy , unrelated Japanese women each gave birth to a child with severe NAIT . STUDY DESIGN AND METHODS : To elucidate the maternal causes of NAIT , we conducted serologic and genetic studies in these two NAIT infants . RESULTS : The serologic experiments localized the antigens to the glycoprotein ( GP ) IIIa subunit of the P08514 /IIIa complex . Sequence-based typing studies subsequently identified a G > A mutation at Nucleotide 1960 ( a glutamic acid > lysine substitution at Position 628 ) in the 11th exon of the P05106 gene . This mutation was recently identified in a report as Q9Y251 -21bw . Next , it was determined that the cause of NAIT in both cases was the Q9Y251 -21bw antigen , as shown by the mothers ' antibodies reacting with the mutated P05106 -transfected cells , but not with transfectants expressing wild-type P05106 . A molecular genetic screening for the Q9Y251 -21bw allele among Japanese donors showed that its genetic frequency in the population was 0.53 % ( 10/1888 ) , indicating that Q9Y251 -21bw occurs at a low but appreciable frequency in the population . Furthermore , in a retrospective study of 50 previous NAIT cases of unknown causes , we found one NAIT case associated with the Q9Y251 -21bw antibody . The two NAIT cases in this study represent the first ones to be associated with Q9Y251 -21bw in Japan . CONCLUSION : We identified the Q9Y251 -21bw allele from two unrelated Japanese infants with severe NAIT . We identified 10 individuals ( 1.06 % ) positive for the Q9Y251 -21bw allele from a genetic screening of 944 Japanese blood donors . Rapid and sustained coronary artery recanalization with combined bolus injection of recombinant tissue-type plasminogen activator and monoclonal antiplatelet P08514 /IIIa antibody in a canine preparation . The effects of bolus injections of recombinant single-chain tissue-type plasminogen activator ( rt-PA ) and of F(ab')2 fragments of a murine monoclonal antibody ( DB00054 ) against the human platelet P08514 /IIIa receptor [ 7E3-F(ab')2 ] on coronary arterial thrombolysis and reocclusion was studied in a canine preparation of coronary artery thrombosis superimposed on high-grade stenosis . Bolus intravenous injections of rt-PA at a dose of 0.45 mg/kg , repeated at 15 min intervals until reperfusion occurred ( maximum of four injections ) caused reperfusion in five of seven dogs within 100 min ( 33 +/- 15 min , mean +/- SD ) . Reperfusion was rapidly followed ( generally within 10 min ) by reocclusion and then by periods of cyclical reflow and reocclusion . A single intravenous injection of 7E3-F(ab')2 alone at 0.8 mg/kg caused reperfusion within 100 min in two of six dogs ( 19 and 37 min ) without subsequent reocclusion . Single bolus injections of different amounts ( 0.1 to 0.8 mg/kg ) of 7E3-F(ab')2 were then combined with bolus injections of 0.45 mg/kg of rt-PA . Stable reperfusion without reocclusion was accomplished with 0.8 or 0.6 mg/kg 7E3-F(ab')2 and a single injection of 0.45 mg/kg rt-PA within 6 +/- 3 min ( n = 6 , p less than .01 ) and 8 +/- 5 min ( n = 5 , p less than .02 ) , respectively . None of these animals suffered reocclusion of the coronary artery . Lower doses ( 0.1 to 0.2 mg/kg ) of 7E3-F(ab')2 did not significantly shorten the time to reperfusion and did not prevent reocclusion. ( ABSTRACT TRUNCATED AT 250 WORDS ) Anti-thrombotic and anticoagulant treatment in interventional cardiology . Efforts to improve Percutaneous Transluminal Coronary Angioplasty ( PTCA ) have resulted in the usage of new antiplatelets , and antithrombotic agents . These new agents may increase bleeding complications . However , EPIC , EPILOG and CAPTURE trials showed benefits of DB00054 , a P08514 /IIIa platelets receptor blocker , in high risk PTCA patients . On the other hand , direct thrombin inhibitors , Hirudin and DB02351 , did not clearly show any benefit when compared to heparin in patients with unstable angina undergoing PTCA . Combination of oral antiplatelets , ticlopidine and aspirin , is widely utilized following stent implantation . However , its benefit over aspirin alone has not been demonstrated . This article aims to review mechanisms and benefits of these new agents in cardiovascular field . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . [ Expression of P11274 / P00519 fusion gene in circulating endothelial cells from chronic myelogenous leukemia patients and its clinical significance ] . Several studies have shown that the tumor endothelial cells are different from the normal tissue endothelial cells . These tumor endothelial cells may contribute to tumor neo-vasculogenesis . This study was purposed to analyze the biologic features and determine the expression level of CD133 and P11274 / P00519 fusion gene in circulating endothelial cells ( CEC ) isolated from peripheral blood of CML patients , as well as to investigate the role of CEC in disease progression . Mononuclear cells were isolated from peripheral blood by density gradient centrifugation ; CEC were sorted by P29966 and harvested in the endothelial growth medium . The morphologic features of CEC were observed by microscopy , the cell growth rate was calculated by cell counting , and the cells were identified by immunofluorescence staining for the expression of CD31, P28906 , P04275 and CD133 . The expression of P11274 / P00519 fusion gene was examined by Q5TCZ1 in 12 CML patients . The results indicated that the isolated CEC displayed the typical cobble-stone morphology . These cells could be identified by the positive immunofluorescence staining for CD31 , P28906 and P04275 , and showed more increased proliferative potential as compared to that of healthy donors . It was found that the positive rate of CD133 was 31.29 % in CML patients , which was significantly different from that of healthy donors ( P < 0.05 ) . In 12 CML patients , CEC carried the same chromosome aberration as the leukemia cells ( 10.77 % ) . Higher expression level of CD133 and P11274 / P00519 fusion gene positively correlated with progression of disease . It is concluded that the CEC may participate in invasion and angiogenesis in patients with CML and possibly correlate to the spreading and progression of the disease . New antithrombotics for the treatment of acute and chronic arterial ischemia . The established antithrombotic agents are effective but they have limitations which have provided opportunities for the development of new antithrombotic compounds . Of these new agents , the antithrombin III-independent thrombin inhibitors and the platelet P08514 /IIIa receptor antagonists are the most advanced in their development . Other new antithrombotic agents include the antithrombin III-independent factor Xa inhibitors , activated protein C , soluble thrombomodulin and tissue factor pathway inhibitor . Of the P08514 /IIIa antagonists , the humanized DB00054 and integrin have been evaluated in phase III studies . The DB00054 was effective in preventing both short-term and longer-term complications of coronary angioplasty . The antithrombin III-independent thrombin inhibitors hirudin and hirulog have also been evaluated in phase III studies . The studies with hirudin as an adjuvant to coronary thrombolysis had to be terminated and restarted at lower dosages because of an unacceptable incidence at intracranial hemorrhage and the study with hirulog produced equivocal results . Genetic polymorphisms in diabetes : influence on therapy with oral antidiabetics . Due to new genetic insights , etiologic classification of diabetes is under constant scrutiny . Hundreds , or even thousands , of genes are linked with type 2 diabetes . Three common variants ( Lys23 of Q14654 , Pro12 of P37231 , and the T allele at rs7903146 of Q9NQB0 ) have been shown to be predisposed to type 2 diabetes mellitus across many large studies . Individually , each of these polymorphisms is only moderately predisposed to type 2 diabetes . On the other hand , monogenic forms of diabetes such as MODY and neonatal diabetes are characterized by unique clinical features and the possibility of applying a tailored treatment.Genetic polymorphisms in drug-metabolizing enzymes , transporters , receptors , and other drug targets have been linked to interindividual differences in the efficacy and toxicity of a number of medications . Mutations in genes important in drug absorption , distribution , metabolism and excretion ( ADME ) play a critical role in pharmacogenetics of diabetes.There are currently five major classes of oral pharmacological agents available to treat type 2 diabetes : sulfonylureas , meglitinides , metformin ( a biguanide ) , thiazolidinediones , and α-glucosidase inhibitors . Other classes are also mentioned in literature.In this work , different types of genetic mutations ( mutations of the gene for glucokinase , HNF 1α , HNF1β and Kir6.2 and Q09428 subunit of KATP channel , Q07869 -γ , OCT1 and OCT2 , cytochromes , direct drug-receptor ( Q14654 ) , as well as the factors that influence the development of the disease ( Q9NQB0 ) and variants of genes that lead to hepatosteatosis caused by thiazolidinediones ) and their influence on the response to therapy with oral antidiabetics will be reviewed . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . [ Functional significance of P25942 and P29965 linking on human lung cells ] . OBJECTIVE : To determine the possible functional significance of P25942 and P29965 ( CD154 ) linking on human lung carcinomas and to assess the potential of P25942 as a therapeutic target . METHODS : We evaluated the effect of P29965 on the surface expression of major histocompatibility complex class I ( MHC-I ) , Fas , bcl2 , CD54 , epidermal growth factor receptor ( P00533 ) , p-glucoprotein ( A6NDG6 ) , lung related protein ( Q14764 ) , cell cycle , cell apoptosis and growth kinetics of 7 lung cancer cell lines ( including 1 P25942 -transfected cell line GLC-82/ P25942 ) by gene cloning , Western blotting , and flow cytometry etc . RESULTS : Significant increased expression of MHC-I , CD54 and Fas was observed in 4 tumor lines expressing high levels of P25942 after P29965 ( 0.1 micro g/ml ) linked to P25942 . P29965 ( 0.1 micro g/ml or greater ) in the fifth day was found to significantly inhibit the proliferation of 4 cell lines expressing high levels of P25942 , decreased the percentage of aneuploid cell , and inhibited S-phase cells entering G2/M phase . The effect of P25942 cross-linking was reversible . P25942 -moderate and low and negative tumor did not respond to P29965 . All of 7 cell lines show no significant changes in apoptosis in either the experimental ( P29965 pulsed ) or control ( medium only ) cell cultures . CONCLUSION : Expression of P25942 on lung tumors cells expressing high level of P25942 ( including P25942 -transfected cells ) may represent a potential therapeutic target . Enhancement of fibrinolysis by gel-filtered platelets and its quenching by cytochalasin B and P08514 /IIIa antagonists . The effects of gel-filtered platelets on euglobulin clot lysis time ( ECLT ) were analyzed to elucidate the possible role of platelets in thrombolysis . Gel-filtered platelet-supplemented ECLT ( plt-ECLT ) was significantly shorter than ECLT without platelets ( regular ECLT ) . DB00054 , anti-glycoprotein IIb/IIIa ( P08514 /IIIa ) antibody , and cytochalasin B nullified the enhancement of ECLT by platelets , and increased plt-ECLT beyond regular ECLT . When gel-filtered platelets were used after disruption , ECLT was not shortened but rather became longer than regular ECLT , probably due to natural fibrinolysis inhibitors released from platelets . Therefore , for platelets to enhance fibrinolysis , intact cell structure and cytoskeletal reorganization after thrombin stimulation is required . Various P08514 /IIIa antagonists prolonged plt-ECLT . The concentrations of P08514 /IIIa antagonists required to prolong plt-ECLT , were varied . Interestingly , the effects of these antagonists were independent of their ability to inhibit thrombin-induced platelet aggregation , but dependent on their ability to induce clot retraction . T-250 , a P08514 /IIIa antagonist , had the smallest effect on plt-ECLT . These drugs do not affect regular ECLT or tissue plasminogen activator ( tPA ) -catalyzed DB00142 -plasminogen activation in the presence of thrombin-activated platelets . Although their overall effect on thrombolysis is inhibitory , platelets could promote fibrinolysis through a P08514 /IIIa-dependent mechanism . A chimeric murine/human antibody Fab fragment directed against the platelet P08514 /IIIa receptor enhances and sustains arterial thrombolysis with recombinant tissue-type plasminogen activator in baboons . Inhibition of the platelet glycoprotein ( GP ) IIb/IIIa receptor with the murine monoclonal antibody 7E3 abolishes ex vivo platelet aggregation , reduces thrombogenicity , and sustains arterial recanalization with recombinant tissue-type plasminogen activator ( rt-PA ) . A chimeric murine/human Fab fragment of DB00054 ( c7E3-Fab ) has a markedly reduced immunogenicity , but its potency as an adjunct for thrombolysis with rt-PA has not been evaluated . The effects of a single intravenous bolus injection of aspirin ( 17 mg/kg ) or c7E3-Fab ( 0.45 mg/kg ) on thrombolysis and reocclusion induced with rt-PA were studied in groups of six baboons with femoral arterial thrombosis and superimposed high-grade stenosis . This dose of c7E3-Fab blocked 96 +/- 1 % of the platelet P08514 /IIIa receptors and abolished ADP-induced platelet aggregation . Bolus intravenous injections of rt-PA ( 0.25 mg/kg ) were repeated at 15-minute intervals until reperfusion occurred ( maximum of four injections ) . In the aspirin group , reperfusion was obtained within 51 +/- 16 minutes ( mean +/- SD ) but was rapidly followed by reocclusion within 6 +/- 9 minutes and by cyclic reflow and reocclusion . In the c7E3-Fab group , reperfusion was obtained within 25 +/- 8 minutes ( P < .01 versus aspirin group ) and was associated with a delayed reocclusion of 63 +/- 63 minutes ( P < .05 versus aspirin group ) . Template bleeding times remained unchanged in the aspirin/rt-PA group but were markedly prolonged ( to > 30 minutes ) in the c7E3-Fab/rt-PA group. ( ABSTRACT TRUNCATED AT 250 WORDS ) Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Therapeutic Implications of a Specific Murine Monoclonal Antibody ( DB00054 ) to the Platelet Receptor P08514 /IIIa . The murine monoclonal antibody 7E3 blocks the platelet glycoprotein IIb/IIIa receptor , and is a potent inhibitor of platelet function in both animals and man . Animal models of the acute coronary syndromes suggest that 7E3 abolishes the in vivo formation of platelet thrombi , accelerates thrombolysis with tissue plasminogen activator , and prevents subsequent reocclusion . Human studies with 7E3 suggest that complete inhibition of platelet function may be safely undertaken for periods of up to 36 h , and preliminary studies indicate effectiveness in the therapy of clinical unstable angina . Potential problems with its use include immunogenicity and thrombocytopenia . The outcome of the acute coronary thrombotic syndromes , which include unstable angina , acute myocardial infarction and abrupt closure following coronary angioplasty , may be significantly improved with 7E3 therapy . Glycoprotein IIb/IIIa inhibition attenuates platelet-activating factor-induced platelet activation by reducing protein kinase C activity . Glycoprotein (GP)IIb/IIIa inhibition may abolish activated leukocyte-induced platelet activation , in which leukocyte-released platelet-activating factor ( Q15004 ) is a major mediator . The present study thus investigated if and how P08514 /IIIa inhibitors interfere with Q15004 -induced platelet activation . Platelet and leukocyte activation were monitored by flow cytometry and immunoblotting . P08514 /IIIa inhibitors ( c7E3 , non-peptide SR121566 , and MAb RFGP56 ) attenuated Q15004 -induced , but not adenosine diphosphate ( ADP ) - or thrombin receptor activating peptide ( TRAP ) -induced platelet P16109 expression in whole blood . P08514 /IIIa blockade enhanced ADP- or TRAP-induced leukocyte CD11b expression , but not the response to Q15004 . P08514 /IIIa blockade attenuated Q15004 -induced , but enhanced ADP- or TRAP-induced platelet-leukocyte aggregation . Under the present experimental conditions , thromboxane A2 receptor antagonism did not significantly influence Q15004 -induced platelet activation , and P08514 /IIIa inhibition did not interfere with calcium mobilization/influx in platelets . Protein kinase C ( PKC ) blockade inhibited Q15004 -induced platelet P16109 expression , and Q15004 -induced PKC activity was reduced by P08514 /IIIa inhibition . Q15004 ( =1 micro m ) did not induce Q02750 /2 or P29323 1/2 phosphorylation , whilst thrombin induced marked responses , which were enhanced by P08514 /IIIa blockade . Thus , P08514 /IIIa inhibition attenuates Q15004 -induced platelet activation via inhibiting PKC activity . P08514 /IIIa blockade enhances thrombin-induced platelet Q02750 /2 and P29323 1/2 activation , and augments ADP- and TRAP-induced leukocyte activation by enhancing platelet-leukocyte aggregation . Regulation of megakaryocytopoiesis by thrombopoietin and stromal cells . P40225 ( Tpo ) is a cytokine which stimulates megakaryocyte maturation . We found that Tpo is constitutively and ubiquitously expressed in all tissues examined , including bone marrow stromal cells , even in thrombocytopenia , thrombosis and steady-state condition in mice . Thus , platelet level in circulation is not regulated by Tpo gene expression . Furthermore , when the purified megakaryocytes were cocultured with the stromal cells , most of the megakaryocytes adhered to the stromal cells and remained unchanged , while free megakaryocytes induced proplatelet formation . Thus the stromal cells in bone marrow secrete Tpo and stimulate megakaryocytopoiesis , but the interaction of megakaryocytes with the stromal cells may suppress platelet formation . Study on signal transduction through Q9UHA4 revealed that Tpo induces activation of O60674 and Tyk2 , which in turn activate P42224 , P40763 and P42229 . Further , Tpo stimulates transcription factors P15976 and NF-E2 , which induce differentiation markers , P08514 /IIIa and Pm-1 . In addition , Shc , Vav , Ras , P04049 , MAPKK , MAPK and Pim-1 are also activated . Thus , Tpo activates a lineage-specific cascade as well as a specific JAK- P35610 cascade and a common signaling cascade . A novel class of 5- Q13049 receptor antagonists : aryl aminoguanidines . Local delivery of serotonin ( 5-HT ) produces a rapid edematous response in soft tissues via increased fluid extravasation which is prevented by 5-HT2 antagonists such as ketanserin or mianserin . Here we report the effects of a new class of aminoguanidine 5-HT2 antagonists , with relative selectivity for 5- Q13049 receptors which are potent inhibitors of 5-HT-induced paw edema in the rat . Radioligand binding studies with 125I DOI on human 5- Q13049 and P28335 receptors and with 3H-5-HT on human P41595 receptors demonstrated that , LY314228 , and LY320954 displayed some selectivity for the 5- Q13049 receptor . When compared to binding at other 5-HT2 receptor subtypes , LY314228 had an 18.6-fold greater affinity for the 5- Q13049 site over the P41595 site , and 2.6 fold greater at the P28335 site . LY320954 displayed similar preference for 5- Q13049 sites . Both compounds also inhibited 5-HT-induced paw swelling in rats , with ED50 's of 6.4 and 4.8 mg/kg ( for LY314228 and LY320954 , respectively ) . These studies offer evidence for a novel class of pharmacophores for the 5-HT2 receptor family which show greater relative affinities for the 5- Q13049 receptor subclass . [ P08514 -IIIa inhibitors ] . Therapy involving the use of anti- P08514 -IIIa inhibitors has progressively evolved in recent years for patients undergoing percutaneous coronary intervention or with acute coronary syndromes . Patients receiving anti-GP IIb-IIIa therapy have a lower risk of death or myocardial infarction than those receiving the classic anti-agregant , aspirin , alone . Two classes of products have been used in clinic , the chimeric monoclonal antibody Fab fragment , c7E3 or abciximab ( ReoPro ) , which has been the pioneer , and synthetic peptides or peptidomimetics such as eptifibatide ( Integrilin ) or tirofiban ( Agrastat ) . DB00054 is a long-acting , high-affinity receptor blocker , whereas eptifibatide and tirofiban have much shorter biological half-lives . Another property that differentiates these compounds is that the peptides bind exclusively to GP IIb-IIIa whereas c7E3 also binds to alpha v beta 3 , the vitronectin receptor . The potent inhibitory effect of these compounds increases the risk of bleeding . By carefully controlling the levels of heparin and by removing the sheath as early as possible , the hemorrhagic problems may be limited . Another potential complication is the rapid development of thrombocytopenia . The cause has yet to be found and for c7E3 no correlation with the development of HACA ( human anti-chimeric antibodies ) has been observed . Because of the chronic nature of coronary artery disease , evaluation of the readministration of c7E3 to the same patient two or even more times is under investigation . The first results do not show major problems . The best biological way to investigate the efficiency of anti- P08514 -IIIa has to be determined . Interestingly , a new point-of-care test has been proposed , while monoclonal antibodies are available that differentiate between nonoccupied and occupied P08514 -IIIa complexes . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . Dynamics of P08514 /IIIa-mediated platelet-platelet interactions in platelet adhesion/thrombus formation on collagen in vitro as revealed by videomicroscopy . The conventional description of platelet interactions with collagen-coated surfaces in vitro , based on serial static measurements , is that platelets first adhere and spread to form a monolayer and then recruit additional layers of platelets . To obtain dynamic information , we studied gravity-driven platelet deposition in vitro on purified type 1 collagen by video phase-contrast microscopy at 22 degrees C . With untreated human and wild-type mouse platelets , soon after the initial adhesion of a small number of " vanguard " platelets , " follower " platelets attached to the spread-out vanguard platelets . Follower platelets then adhered to and spread onto nearby collagen or over the vanguard platelets . Thus , thrombi formed as a concerted process rather than as sequential processes . Treatment of human platelets with monoclonal antibody ( mAb ) DB00054 ( anti- P08514 /IIIa ( alphaIIbbeta3 ) + alphaVbeta3 ) or tirofiban ( anti- P08514 /IIIa ) did not prevent platelet adhesion but nearly eliminated the deposition of follower platelets onto vanguard platelets and platelet thrombi . Similar results were obtained with Glanzmann thrombasthenia platelets . Wild-type mouse platelets in the presence of mAb 1B5 ( anti- P08514 /IIIa ) and platelets from beta3-null mice behaved like human platelets in the presence of 7E3 or tirofiban . Deposition patterns of untreated human and wild-type mouse platelets were consistent with random distributions under a Poisson model , but those obtained with 7E3- and tirofiban-treated human platelets , 1B5-treated mouse platelets , or beta3-null platelets demonstrated a more uniform deposition than predicted . Thus , in this model system , absence or blockade of P08514 /IIIa receptors interferes with thrombus formation and alters the pattern of platelet deposition . P08908 receptor responsivity in unipolar depression . Evaluation of ipsapirone-induced DB01285 and cortisol secretion in patients and controls . The selective P08908 receptor ligand ipsapirone ( IPS ) induces corticotropin ( DB01285 ) and cortisol secretion in humans . To explore P08908 receptor-mediated hypothalamic-pituitary-adrenal ( Q9Y251 ) system activation in depression , 24 subjects ( 12 patients with unipolar depression and 12 individually matched controls ) were given 0.3 mg/kg IPS or placebo in random order . Compared with controls , the depressed patients exhibited significantly decreased DB01285 and cortisol responses to IPS in association with increased basal cortisol secretion . The impaired Q9Y251 response following P08908 receptor challenge in unipolar depression could have resulted from glucocorticoid-dependent subsensitivity of the ( post-synaptic ) P08908 receptor itself and/or from a defective postreceptor signaling pathway [ inhibitory guanine nucleotide-binding protein ( Gi ) -adenylate cyclase complex function ] , thus supporting the hypothesis that a disintegrated 5-HT and Q9Y251 system interaction may be present in depression . Future studies of the Q9Y251 response to direct-acting P08908 ligands , such as IPS , should facilitate the assessment of 5-HT/ Q9Y251 system integrity in various affective disorders and its involvement in psychotropic drug effects . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Investigation of interaction of human platelet membrane components with anticoagulant drugs DB00054 and DB00063 . DB00054 ( Abci ) and eptifibatide ( Epti ) are antiaggregate drugs which may reduce thrombotic complications in acute coronary syndromes . The aim of this work was the investigation of the interaction between the phospholipid- P08514 /IIIa glycoprotein complex and Abci or Epti , and the influence of these drugs on the phospholipid ratio in the platelet membrane . The interaction between the phospholipid- P08514 /IIIa glycoprotein complex and antiaggregate drugs were investigated using the Surface Plasmon Resonance Imaging technique ( SPRI ) . Phospholipids phosphatidylinositol ( PI ) , phosphatidylserine ( PS ) , phosphatidylethanolamine ( PE ) , phosphatidylcholine ( PC ) and sphingomyelin ( SM ) were first immobilized onto the gold chip surface . The phospholipid ratio in the platelet membrane was determined by the HPLC . Only PI , PS , PE and PC were determined . Human platelets treated ' in vitro ' with Abci or Epti exhibit changes in the phospholipid ratio in the platelet membrane . The ratio of PS decreases and PC rises . The SPRI distinctly shows interactions between phospholipids and glycoprotein P08514 /IIIa , and between the phospholipid-glycoprotein P08514 /IIIa complex and Abci or Epti . The interaction between phospholipids and glycoprotein P08514 /IIIa is growing in the sequence : PI << SM < PE < PC < PS . The interaction between phospholipid-glycoprotein P08514 /IIIa complex and Abci/Epti is growing in the sequence : PS < PI < PC < PE < SM . SPRI was proved to be excellent tool for observation of such interactions . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . A new short chain RGD-containing disintegrin , accutin , inhibits the common pathway of human platelet aggregation . A new short-chain disintegrin , accutin , was purified from the Formosan Agkistrodon acutus venom by using of gel filtration , ion exchanger and reverse phase HPLC . The homogeneous protein is a 47-residue polypeptide with a molecular mass of 5241 Da containing an DB00125 - DB00145 - DB00128 sequence and seven cysteinyl residues at positions highly homologous to other disintegrins . Accutin dose-dependently inhibited human platelet aggregation stimulated by ADP , collagen , thrombin or the thromboxane analogue U46619 in platelet suspension with IC50 values of 66-267 nM . It was also active in inhibiting platelet aggregation of platelet-rich plasma . However , accutin apparently did not affect the shape change caused by these agonists . Accutin also inhibited fibrinogen-induced aggregation of human elastase-treated platelets in a dose-dependent manner . Furthermore , accutin dose-dependently inhibited the binding reaction of fluorescein isothiocyanate ( FITC ) -conjugated arietin , a member of the disintegrin family , to human platelets . In addition , the binding of FITC-conjugated accutin to platelets was almost completely blocked by a monoclonal antibody , DB00054 , raised against the platelet glycoprotein IIb/IIIa complex . On the other hand , accutin as well as other disintegrins , rhodostomin and arietin , exhibited an inhibitory effect on 7E3 binding toward platelets and endothelial cells in a dose-dependent manner . It is concluded that accutin , a new platelet aggregation inhibitor belonging to the short-chain disintegrin family , acts specifically on a binding epitope of P08514 /IIIa overlapping with that of DB00054 , leading to the blockade of fibrinogen binding to its receptor . The effect of s-nitroso-glutathione on platelet and leukocyte function during experimental extracorporeal circulation . Treatment with extracorporeal membrane oxygenation ECMO ) is associated with side effects , e.g. , blood cell consumption and activation . Our group has earlier shown that nitric oxide administered as a gas reduces platelet consumption and activation . In the present work we have studied the effect of the NO-donor S-nitroso-glutathione GSNO ) on platelets and leukocytes in an in vitro extracorporeal circuit . Two complete ECMO circuits were perfused with fresh heparinized human blood for 24 hours . GSNO was administered as a continuous infusion to one circuit at a rate of 0.7 mg/hour in four paired experiments and at a rate of 3.5 mg/hour in another four paired experiments . The other circuit was used as a control . Blood samples were withdrawn from both circuits before the start of the experiments and at 0.5 , 1 , 3 , 12 , and 24 hours of perfusion . The samples were analyzed for red blood cell count , leukocyte count , platelet count , platelet membrane expression of glycoproteins GP ) Ib and P08514 /IIIa , leukocyte membrane expression of cluster of differentiation CD ) 11b/ P05107 , as well as plasma concentration of tumor necrosis factor P01375 ) -alpha , interleukin IL ) -1beta , and P10145 . No difference in these parameters between the GSNO and the control circuit at any time point was assayed . In this study , no significant effect of GSNO on circulating platelets or leukocytes during experimental extracorporeal circulation could be shown . Q13444 is an adhesion receptor for platelet P08514 -IIIa and induces platelet activation . Cell adhesion and proteolytic matrix degradation are central processes in atherosclerosis . Being a member of the family of ADAMs ( " a disintegrin and metalloproteinase " ) , metargidin ( Q13444 ) combines a metalloproteinase domain and an RGD aminoacid sequence . We studied the potential role of Q13444 as an adhesion receptor on endothelial cells and interactions between platelets and Q13444 with respect to platelet adhesion , activation and thrombus formation . Q13444 was found to be expressed on cultured endothelial cells ( HUVEC ) . Platelet adhesion to immobilized recombinant Q13444 was effectively enhanced under both static and high shear rate conditions reaching the maximum level of adhesion to fibrinogen . Consistently , platelet adhesion onto Q13444 overexpressing endothelial cells was significantly increased . Adhesion to Q13444 was reduced by blockade of P08514 -IIIa using neutralizing anti-alpha(IIb)beta3 mAbs ( DB00054 , 2G12 ) , but not by anti-alpha(v)beta3 ( LM609 ) . Soluble Q13444 binds to activated but not to resting P08514 -IIIa . Moreover , platelets adherent to Q13444 additionally attracted platelets under high shear rates indicating an initial role of platelet- Q13444 interactions for thrombus formation . Furthermore , incubation of platelets with soluble Q13444 showed a dose-dependent increase in secretion of CD62P and P29965 . Q13444 is expressed on endothelial cells and can serve as an adhesion receptor for platelets via P08514 -IIIa binding . Platelet adhesion to Q13444 leads to platelet activation , secretion and promotes thrombus formation . Thus , Q13444 may represent a novel target for antithrombotic strategies in cardiovascular pathologies . Inhibition of angiogenesis and tumor growth by murine DB00054 , the parent antibody of c7E3 Fab ( abciximab ; ReoPro ) . Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers . The expression of endothelial cell integrin alpha(v)beta3 has been shown to increase during vascular proliferation associated with human tumors . Selective antagonists of alpha(v)beta3 can block angiogenesis and tumor growth by inducing programmed cell death in proliferating endothelial cells . Monoclonal antibody DB00054 , an antagonist of the human , but not murine , integrins alpha(v)beta3 and alphaIIbbeta3 ( P08514 /IIIa ) , inhibits platelet aggregation . It is the parent antibody of a mouse/human chimeric antibody fragment approved for adjunctive therapy of patients undergoing percutaneous coronary interventions to prevent ischemic complications ( c7E3Fab ; abciximab ; ReoPro ) . To evaluate the potential of 7E3 to inhibit human angiogenesis and tumor growth independent of its antiplatelet effects , we established integrin alpha(v)beta3-negative human melanoma tumors in full-thickness human skin grafted onto SCID mice . The resulting tumors induce a human angiogenic response as assessed by the immunoreactivity of vascular cells with monoclonal antibodies specific for human CD31 . Administration of 7E3 prevented or significantly inhibited the growth of tumors , and this effect correlated with a significant reduction in the number of blood vessels supplying the tumors . These results support the previous findings that blockade of integrin alpha(v)beta3 inhibits angiogenesis and tumor growth and indicates that dual inhibitors of alpha(v)beta3 and alphaIIbbeta3 are effective in blocking tumor growth and angiogenesis . Expression of growth factors and growth factor receptor in non-healing and healing ischaemic ulceration . OBJECTIVES : To characterise the histological and cytokinetic characteristics of purely ischaemic ulcers and the processes that underpin healing following successful revascularisation . DESIGN : Prospective observational study . MATERIALS AND METHODS : Biopsies were taken immediately pre- and 6 weeks following successful revascularisation of solely ischaemic ulceration . They were evaluated for morphological differences using H & E staining for the platelet derived growth factor receptor ( P09619 ) , epidermal growth factor receptor ( P00533 ) , TGFbeta receptorIII ( TGFbetaRIII ) , transforming growth factor beta 1 and 3 ( TGFbeta1 and TGFbeta3 ) and P04275 ( P04275 ) expression using immunohistochemistry . Localisation and quantification of these growth factors and receptors was assessed systematically by three independent investigators who were blinded to the timing of biopsy . RESULTS : Pre-operatively , small vessel vasculitis , necrosis and infection with a profuse neutrophil and macrophage infiltrate was observed in all samples . Post-operative biopsies revealed a proliferation of new capillaries in and around the ulcer edge and base . P04275 staining confirmed an endothelial layer within these new vessels . Following successful revascularisation there was less infection and inflammation with minimal vasculitis . These newly formed capillaries had increased staining for TGFbeta3 , P09619 and TGFbetaRIII with staining for P09619 also localised to dermal fibroblasts which were larger and more numerous . Accelerated epithelial cell proliferation was observed with detachment from the underlying dermis . CONCLUSIONS : Healing of purely ischaemic ulcers is characterised by vasculogenesis associated with increased presence of the proangiogenic cytokines PDGF and TGFbeta3 . These findings show promise for the use of growth factor manipulation to aid healing in ischaemic ulcers . New antiplatelet agents : platelet P08514 /IIIa antagonists . The P08514 /IIIa ( alpha IIb beta 3 ) receptor plays a crucial role in platelet aggregation and platelet thrombus formation . Inhibition of P08514 /IIIa with the Fab fragment of the mouse/human chimeric monoclonal antibody DB00054 , snake venom peptides containing the arginine-glycine-aspartic acid ( RGD ) sequence , or peptides or peptidomimetics based on the RGD sequence results in abolition of platelet aggregation and platelet thrombus formation . This results in profound inhibition of thrombotic occlusions in animal models . The Phase III EPIC study demonstrated that c7E3 Fab , given as bolus followed by a 12 h infusion , reduced the risk of acute ischemic complications after coronary angioplasty by approximately 35 % in patients at high risk of suffering such complications . Treated patients had an approximately 2-fold increased risk of major bleeding , but no increase in cerebral hemorrhage or lethal bleeding . Treatment with c7E3 Fab may have had a beneficial effect on clinical restenosis at 6 months , but this needs to be confirmed . A possible anticoagulant effect of c7E3 Fab was also identified in EPIC , and in vitro studies support this possibility . With the approval of c7E3 Fab ( abciximab ; ReoPro ) for patients undergoing high-risk angioplasty in the US and several European and Scandinavian countries , P08514 /IIIa inhibition joins the armamentarium of antithrombotic agents . No evidence for an influence of the human platelet antigen-1 polymorphism on the antiplatelet effects of glycoprotein IIb/IIIa inhibitors . This study investigated the hypothesis that the human platelet antigen-1 ( Q9Y251 -1 ) polymorphism may influence the antiplatelet effects of glycoprotein (GP)IIb/IIIa inhibitors . DB00640 diphosphate ( 30 micro mol ) -induced fibrinogen binding was measured by flow cytometry . DB00054 ( 0.03-3 micro g/ml ) , tirofiban ( 0.3-30 nmol/l ) or eptifibatide ( 0.01-1 micro g/ml ) were incubated for 15 min with the samples prior to stimulation . IC(50) values for the inhibition of fibrinogen binding were determined from each experiment . All subjects were genotyped by GALIOS and automated fluorescence correlation spectroscopy . Although a marked variability in the inhibitory effects of all three P08514 /IIIa inhibitors was confirmed , there were no significant differences between the genotypes with respect to the inhibition of fibrinogen binding . Thus , the present study does not provide evidence for an effect of Q9Y251 -1 polymorphism on the inter-individual variability in the platelet inhibitory effects of the three P08514 /IIIa inhibitors approved for clinical use . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Potential future clinical applications for the P08514 /IIIa antagonist , abciximab in thrombosis , vascular and oncological indications . DB00054 ( ReoPro ) is a mouse-human chimeric monoclonal antibody Fab fragment of the parent murine monoclonal antibody DB00054 , and was the first of these agents approved for use as adjunct therapy for the prevention of cardiac ischemic complications in patients undergoing percutaneous coronary intervention ( P05154 ) . DB00054 binds with high avidity to both the non-activated and activated form of the P08514 /IIIa receptor of platelets , the major adhesion receptor involved in aggregation . Additional cardiovascular indications for abciximab are unstable angina , carotid stenting , ischemic stroke and peripheral vascular diseases . DB00054 also interacts with two other integrin receptors ; the a av b b3 receptor , which is present in low numbers on platelets but in high density on activated endothelial and smooth muscle cells , and a aMb b2 integrin which is present on activated leukocytes . Cell types that express integrins P08514 /IIIa and a av b b3 such as platelets , endothelial and tumor cells have been implicated in angiogenesis , tumor growth and metastasis . Since abciximab interacts with high avidity to integrins P08514 /IIIa and a av b b3 , it is reasonable to assume that it may possess anti-angiogenic properties in angiogenesis-related diseases , as well as anti-metastastatic properties in case of disseminating tumors expressing the target integrin receptors . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Forced expression of P38936 in P08514 - P38936 transgenic mice induces abnormalities in the proliferation of erythroid and megakaryocyte progenitors and primitive hematopoietic cells . OBJECTIVE : P38936 ( P38936 /Cip/kip) and p27(Kip1) are cyclin-dependant kinase inhibitors controlling cell-cycle exit and differentiation of numerous cell types . Among hematopoietic cells , megakaryocytes express high levels of P38936 , while in erythroid cells , p27(Kip1) is predominant . As P38936 and p27 could display overlapping functions and as megakaryocytes and erythroid cells derive from a bipotent progenitor , we developed an in vivo model to determine the specific role of P38936 in controlling the proliferation/differentiation balance of erythroid and megakaryocytic progenitors . METHODS : Transgenic mice that overexpressed P38936 under the control of the human P08514 promoter in early progenitors and along megakaryocytic differentiation were generated . Different subsets of hematopoietic progenitors ( BFU and CFU ) and primitive cells ( CAFC , LTC-IC ) were analyzed by methylcellulose assay . Phenotypic evolution and clonogenic properties of the lin(-) population were analyzed along erythroid and megakaryocytic differentiation . RESULTS : We observed P38936 ectopic expression in early hematopoietic progenitors ( lin(-)Sca(+) ) , megakaryocytes , and , to a lesser extent , erythroid cells . This expression induced an important decrease in the number of CFU-MK , BFU-E , CFU-E , primitive progenitors ( CAFC day 35 ) , and LTC-IC , but did not affect the maturation process of these cells and the blood cell count . CONCLUSIONS : We show that variation of P38936 expression level changes the fate of hematopoietic cells by favoring either proliferation or differentiation pathways . This effect of P38936 is exerted not only at the level of primitive progenitors but also in more mature progenitors . However , in vivo , a systemic compensation mechanism is most likely activated in response to variations of the flow of progenitor production . DB00054 pharmacodynamics are unaffected by antecedent therapy with other P08514 /IIIa antagonists in non-human primates . BACKGROUND : Tirofiban and eptifibatide are currently approved for the medical stabilization of non-ST segment elevation acute coronary syndromes . In patients undergoing percutaneous coronary intervention ( P05154 ) during infusion of these drugs , conversion to abciximab , which has long term proven clinical efficacy and cost-effectiveness , following P05154 may be desirable . The purpose of this study was to determine if the binding or pharmacodynamics of abciximab is affected by a prior infusion of either tirofiban or eptifibatide . METHODS : In vitro binding experiments were performed to determine if prior exposure to tirofiban or eptifibatide altered the affinity and extent of binding of abciximab to P08514 /IIIa . For in vivo experiments , cynomolgus monkeys were pretreated with a bolus and 18 hour infusion of saline , tirofiban , or eptifibatide . At the end of the initial treatment , a bolus and 12 hr infusion of abciximab was started without delay . Inhibition of platelet aggregation , P08514 /IIIa receptor blockade and abciximab pharmacokinetics were measured during and after both infusions . RESULTS : Equilibrium binding of abciximab in vitro was unaffected by tirofiban or eptifibatide . The extent and duration of abciximab inhibition of ex vivo platelet aggregation , receptor blockade , and abciximab pharmacokinetics in monkeys during and after the abciximab infusion were not affected by prior infusion of the animals with tirofiban or eptifibatide . CONCLUSIONS : In vitro and in vivo studies revealed that the molecular interaction of abciximab with the platelet P08514 /IIIa receptor is not altered by immediate prior exposure of platelets to small molecule P08514 /IIIa antagonists . These preclinical studies suggest that the efficacy of abciximab should not be impaired if it is initiated following termination of therapy with small molecule P08514 /IIIa antagonists . Monoclonal antibody against the platelet glycoprotein ( GP ) IIb/IIIa receptor prevents coronary artery reocclusion after reperfusion with recombinant tissue-type plasminogen activator in dogs . Localized thrombosis was produced in the left anterior descending ( LAD ) coronary artery of open chest dogs by constricting a segment so as to produce greater than 90 % stenosis ( reducing blood flow to 40 +/- 10 % of baseline ) , and placing a thrombus in the segment immediately proximal to the stenosis by inducing endothelial cell injury and instilling a mixture of blood and thrombin . Intravenous infusion of recombinant tissue-type plasminogen activator ( rt-PA ) at a rate of 15-30 micrograms/kg per min for 30 or 60 min in eight dogs induced coronary artery reperfusion within 23 +/- 7 min ( mean +/- SD ) , but reocclusion occurred despite heparin anticoagulation in all but one of these dogs within 7 +/- 5 min . Intravenous injection of 0.8 mg/kg of the F(ab')2 fragment of a monoclonal antibody ( DB00054 ) directed against the platelet P08514 /IIIa receptor , prevented reocclusion in 10/10 dogs during an observation period of 2 h ( P less than 0.001 vs. rt-PA alone ) . The antibody abolished ADP-induced platelet aggregation and markedly prolonged the bleeding time . Intravenous aspirin or dipyridamole prevented reocclusion for 1 h or more in only 2/7 and 1/6 dogs , respectively . We conclude that the monoclonal antibody is very effective in preventing reocclusion after successful thrombolysis of occluded coronary arteries with rt-PA . DB01109 potentiation of collagen-induced platelet aggregation is related to the P08514 / P05106 receptor and not to the GPIb receptor , as tested by whole blood aggregometry . To determine whether heparin potentiation of platelet aggregation is related to platelet GP IIb/IIIa and GP Ib receptors , four series of experiments were performed on blood from normal volunteers . In the first experiment pretreatment with the monoclonal antibody DB00054 ( MAb DB00054 ) , which antagonizes at the GP IIb/IIIa receptor , potently inhibited the collagen-induced platelet aggregation ( p less than 0.001 ) . With heparin added to blood pretreated with MAb DB00054 , the aggregation increased ( p less than 0.005 ) to an extent similar to that when only saline was used for pretreatment . In the second experiment , monoclonal antibody 10E5 ( MAb 10E5 ) and peptide RGDS , substances which also antagonize at the GP IIb/IIIa receptor , decreased collagen-induced platelet aggregation to an extent similar to that after pretreatment with MAb DB00054 . Following pretreatment with RGDS , heparin increased platelet aggregation ( p less than 0.03 ) , while after pretreatment with antibody MAb 10E5 heparin did not enhance platelet aggregation . In the third experiment aurin , an inhibitor of P04275 and its interaction with the platelet GPIb receptor , decreased platelet aggregation dose-dependently . In the fourth experiment heparin enhanced platelet aggregation to a similar extent ( p less than 0.005 ) , regardless of pretreatment of the blood with saline , aurin or monoclonal antibody 6D1 ( MAb 6D1 ) , the latter an antagonist at the GP Ib receptor . In conclusion , the potentiation of collagen-induced platelet aggregation by heparin was not inhibited by MAb DB00054 , RGDS , aurin or MAb 6D1 , but was abolished by MAb 10E5 , implying that the heparin effect is related to activation of the platelet GP IIb/IIIa receptor complex . Highly electronegative LDL from patients with ST-elevation myocardial infarction triggers platelet activation and aggregation . Platelet activation and aggregation underlie acute thrombosis that leads to ST-elevation myocardial infarction ( STEMI ) . Q15004 -highly electronegative low-density lipoprotein ( LDL ) -is significantly elevated in patients with STEMI . Thus , we examined the role of Q15004 in thrombogenesis . Plasma LDL from patients with STEMI ( n = 30 ) was chromatographically resolved into 5 subfractions ( Q9NUQ9 - Q15004 ) with increasing electronegativity . In vitro , Q15004 enhanced adenosine diphosphate-stimulated platelet aggregation twofold more than did Q9NUQ9 and induced platelet-endothelial cell ( EC ) adhesion . Q15004 also increased P16109 expression and glycoprotein (GP)IIb/IIIa activation and decreased cyclic adenosine monophosphate levels ( n = 6 , P < .01 ) in platelets . In vivo , injection of Q15004 ( 5 mg/kg ) into C57BL/6 mice twice weekly for 6 weeks shortened tail bleeding time by 43 % ( n = 3 ; P < .01 vs Q9NUQ9 -injected mice ) and increased P16109 expression and P08514 /IIIa activation in platelets . Pharmacologic blockade experiments revealed that Q15004 signals through platelet-activating factor receptor and lectin-like oxidized P01130 -1 to attenuate Akt activation and trigger granule release and P08514 /IIIa activation via protein kinase C-α . Q15004 but not Q9NUQ9 induced tissue factor and P16109 expression in human aortic ECs ( P < .01 ) , thereby triggering platelet activation and aggregation with activated ECs . These findings indicate that elevated plasma levels of Q15004 may promote thrombosis that leads to STEMI . P10275 is causally involved in the homeostasis of the human prostate endothelial cell . Androgen deprivation causes a reduction of blood flow in the prostate gland that precedes temporally apoptosis of the epithelium . The acute response of prostate endothelial cells to androgen deprivation suggested they represent a primary target for androgen . However , rat prostate endothelial cells were reported not to express androgen receptor ( AR ) , and the role of the androgen axis in human prostate endothelial cell ( HPEC ) homeostasis was poorly characterized . In this study AR expression was detected in HPEC in vivo in clinical specimens of benign prostate and prostate cancer , and AR function as a transcription factor was demonstrated in HPEC in primary xenografts of human benign prostate tissue transplanted into severe combined immunodeficient mice by iv administration of adenoviral mouse mammary tumor virus-driven luciferase expression vector . AR expression and functionality were maintained in vitro in primary cultures of HPEC that coexpressed CD31 , P28906 , P04275 , intercellular adhesion molecule , vascular endothelial growth factor receptor 1 , and vascular endothelial growth factor receptor 2 but did not express prostate-specific antigen . AR expression in primary cultures of HPEC isolated from surgical specimens of benign prostate was validated using RT-PCR , cDNA sequencing , immunocytochemistry , and Western blot analyses . Scatchard analyses demonstrated a single ligand-binding site for R1881 in primary cultures of HPEC , with dissociation constant of 0.25 nm , and AR-mediated transcriptional activity was demonstrated using adenoviral mouse mammary tumor virus-driven luciferase reporters . DB02901 increased proliferation in primary cultures of HPEC in a dose-dependent manner without modulating endothelial tube formation in Matrigel ( BD Biosciences , Bedford , MA ) . Therefore , HPECs express functional AR , and androgen plays a direct role in modulating HPEC biology . P05305 induced proinflammatory markers in the myocardium and leukocytes of guinea-pigs : role of glycoprotein IIB/IIIA receptors . AIM : To assess whether endothelin-1 ( ET-1 ) induces the in vivo expression of inflammatory-related proteins , namely cyclooxygenase-2 ( P35354 ) and tissue factor , in the myocardium and circulating leukocytes of guinea-pigs . The involvement of platelets was also analyzed . METHODS : ET-1 ( 0.013 microg/min ) was infused to male guinea-pigs for 45 min in the presence and absence of tirofiban , a nonpeptidic blocker of the glycoprotein IIb/IIIa receptor ( P08514 /IIIa ) . P13726 and P35354 expression were determined by Western blot . RESULTS : No changes in mean arterial pressure and heart rate were detected . ET-1-infused guinea-pigs showed a marked increase in the number of platelets expressing activated P08514 /IIIa receptors ( 0.8+/-0.03 % vs. 6.5+/-0.2 % ; P < 0.05 ) . Tirofiban ( 10 microg/Kg bw/min ) blunted ex vivo platelet aggregation in response to ADP , although only partially reduced P35354 and tissue factor expression in both the myocardium and leukocytes of ET-1-infused guinea-pigs . The myocardium of platelet-depleted guinea-pigs also showed a reduced P35354 expression after ET-1 infusion ( 57+/-3 % reduction ; P < 0.05 ) . In vitro studies demonstrated that platelets ( 10(7) and 10(9) platelets/well ) enhanced ET-1 ( 10(-7) mol/l ) -induced P35354 expression in heart slices . CONCLUSION : ET-1 stimulated in vivo the expression of the pro-inflammatory proteins P35354 and tissue factor in the myocardium and in leukocytes by a mechanism P08514 /IIIa platelet receptors . P08514 /IIIa antagonists and other anti-integrins . Platelet aggregation involves the binding of adhesive proteins ( fibrinogen , P04275 ) to the alphaIIbbeta3 integrin , which assumes a high-affinity state for adhesive proteins during platelet activation . The occupied integrin sends signals back into the platelet , and the bound adhesive protein forms the bridges linking platelets together . Anti-integrin therapy is designed to inhibit this process in arterial thrombosis . DB00054 , mouse-human chimeric Fab fragments , blocks platelet aggregation and provides proven clinical benefit in acute situations such as in patients with unstable angina undergoing angioplasty or stenting . DB00063 and tirofiban are small molecular mass inhibitors also in current use . In contrast , oral inhibitors of alphaIIbbeta3 have proved disappointing , provoking increased mortality without assuring an adequate blockade of alphaIIbbeta3 . The problems of using anti-integrin therapy are discussed in this article as are ways of improving its efficacity . Final thoughts provide ideas for a new generation of inhibitors . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Conjugation and evaluation of 7E3 x P4B6 , a chemically cross-linked bispecific F(ab')2 antibody which inhibits platelet aggregation and localizes tissue plasminogen activator to the platelet surface . A bispecific F(ab')2 monoclonal antibody which recognizes both the platelet P08514 /IIIa receptor and human tissue plasminogen activator was produced to target tPA to platelets for enhancement of thrombolysis . A stable , thioether-cross-linked bispecific F(ab')2 ( 7E3 X P4B6 ) combining the P08514 /IIIa-specific monoclonal antibody DB00054 , which inhibits platelet aggregation , and a nonneutralizing anti-tPA monoclonal antibody ( P4B6 ) was produced . This was performed by coupling each of the parental Fab ' moieties with the homobifunctional cross-linker bis ( maleimido methyl ) ether ( BMME ) . 7E3 X P4B6 was sequentially purified using gel-filtration chromatography and hydrophobic interaction ( HIC ) HPLC . HIC was shown to completely resolve each of the parental F(ab')2 species from the bispecific one . 7E3 X P4B6 was shown to retain completely each of the parental immunoreactivities in P08514 /IIIa and tPA binding EIA 's . The bispecific antibody inhibited platelet aggregation in vitro at levels comparable to those for 7E3 Fab . Recruitment of tPA activity to washed human platelets was demonstrated using the S-2251 chromogenic substrate assay . 7E3 X P4B6 recruited 12-fold more tPA to the washed platelets than a mixture of the parental F(ab')2 molecules used as controls . Maturation of dendritic cells by recombinant human P29965 -trimer leads to a homogeneous cell population with enhanced surface marker expression and increased cytokine production . Dendritic cells ( DC ) have been shown to be potent inducers of specific cytotoxic T-cell responses both in vivo and in vitro . Furthermore , exposure to cytokines such as tumour necrosis factor ( P01375 ) -alpha or P25942 triggering changes DC phenotype and cytokine production and may enhance the T-cell activating capacity of the DC . We studied DC phenotype and cytokine production as well as the T-cell proliferation and cytotoxic T lympocyte ( CTL ) activation induced by DC generated in vitro . In addition , the effect of exposure to recombinant human P29965 -trimer ( huCD40LT ) on these parameters was investigated . Effective differentiation of monocytes derived from freshly isolated peripheral blood mononuclear cells ( PBMC ) was obtained with granulocyte macrophage-colony stimulating factor ( GM- P04141 ) and interleukin ( IL ) -4 . The DC expression of human leucocyte antigen ( HLA ) molecules , P33681 , Q01151 , and P42081 was markedly enhanced by exposure to huCD40LT even compared to P01375 exposure . Only a moderate cytokine production was observed initially , while P01375 addition or P25942 triggering , especially , induced enhanced production of P05231 and IL-12 p40 . Surprisingly , comparable induction of T-cell proliferation by a DC allostimulus or through the presentation of PPD , and influenza M1-peptide specific CTL activity was obtained with nonmaturated ( Q01151 - ) and maturated ( Q01151 + ) DC . In conclusion , a final maturation of monocyte-derived DC through huCD40LT resulted in a highly homogeneous cell population with enhanced surface marker expression and high production of pro-inflammatory cytokines . In addition , the induction of responses to allo or recall antigens presented by huCD40LT maturated DC was comparable to the responses obtained with the DC maturated through P01375 exposure . The anti- P08514 -IIIa agents : fundamental and clinical aspects . The platelet P08514 /IIIa receptor mediates platelet aggregation induced by all physiologic agonists . Blockade of the receptor , either by monoclonal antibodies or small molecules patterned after the arginine glycine-aspartic acid ( RGD ) cell recognition domain , prevents arterial thrombosis in animal models much better than does aspirin . c7E3 Fab , the Fab fragment of the mouse/human chimeric antibody DB00054 ( abciximab : ReoPro ) , was shown to reduce ischemic events after angioplasty when given in conjunction with heparin and aspirin to patients at high risk in the EPIC study , but its was associated with an increase in bleeding . Preliminary data from the subsequent EPILOG study , in which a lower dose of heparin was used , demonstrated efficacy in low risk as well as high risk patients and no significant increase in major bleeding . Preliminary data from the CAPTURE study support the use of c7E3 Fab in patients with unstable angina who are candidates for PTCA within 24 hours . Positive trends toward decreased thrombotic events have also been observed in patients treated with small molecule inhibitors of P08514 /IIIa receptors . This new class of agents thus holds promise for improving the therapy of angioplasty as well as perhaps other thrombotic phenomena . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . Analysis of P08514 /IIIa receptor number by quantification of 7E3 binding to human platelets . A large number of glycoprotein ( GP ) IIb/IIIa receptors are present on the surface of platelets . Studies to define precisely the number of P08514 /IIIa receptors using specific monoclonal antibodies ( MoAbs ) or fibrinogen binding have , however , yielded varying estimates of receptor number . To refine the quantitative estimation of P08514 /IIIa receptors on resting platelets , we have used the MoAb DB00054 , which has high affinity for P08514 /IIIa . Quantitative binding studies were performed using radiolabeled conjugates of 7E3 IgG , as well as fragments and derivatives of DB00054 . For platelets obtained from any single individual , the numbers of 7E3 F(ab')2 and IgG molecules bound per platelet were equivalent ( approximately 40,000 ) , whereas the number of Fab molecules bound per platelet was consistently approximately twofold higher ( approximately 80,000 ) . To investigate the basis of the quantitative disparity in binding of intact 7E3 and 7E3 F(ab')2 versus 7E3 Fab , we studied the binding of a newly constructed , bispecific (Fab')2 molecule containing only a single 7E3 combining site . Because this construct bound to the same extent as the Fab species , the larger size of the intact 7E3 and 7E3 F(ab')2 molecules could not explain the reduced number of molecules that bound per platelet compared to the Fab fragment . Rather , it appears that the valency of the antibody is the critical factor determining the number of antibody molecules bound per platelet . Thus , we conclude that the binding of 7E3 Fab corresponds most closely with surface P08514 /IIIa number and that the number of P08514 /IIIa receptors is approximately 80,000 per platelet .

MH_train_1011

MH_train_1011

MH_train_1011

interacts_with DB00921?

multiple_choice

Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Serotonin and dopamine receptor gene polymorphisms and the risk of extrapyramidal side effects in perphenazine-treated schizophrenic patients . RATIONALE : DB00850 , a classical antipsychotic drug , has the potential to induce extrapyramidal side effects ( EPS ) . Dopaminergic and serotonergic pathways are involved in the therapeutic and adverse effects of the drug . OBJECTIVES : To evaluate the impact of polymorphisms in the dopamine D(2) and D(3) and serotonin 2A and 2C receptor genes ( P14416 , P35462 , P28223 , and P28335 ) on short-term effects of perphenazine monotherapy in schizophrenic patients . MATERIALS AND METHODS : Forty-seven Estonian inpatients were evaluated before and after 4-6 weeks of treatment by Simpson-Angus rating scale , Barnes scale , and Positive and Negative Symptom Scale . Genotyping was performed for common P14416 , P35462 , P28223 , and P28335 gene polymorphisms , previously reported to influence receptor expression and/or function . RESULTS : Most of the patients ( n = 37 ) responded to the treatment and no significant association was observed between the polymorphisms and antipsychotic response . The 102C allele of P28223 and the -697C and 23Ser alleles of P28335 were more frequent among patients with EPS ( n = 25 ) compared to patients without EPS ( n = 22 ) ( p = 0.02 , 0.01 , and 0.02 , respectively ) . The difference between patients with and without EPS in variant allele frequencies remained significant after multiple model analyses including age , gender , and duration of antipsychotic treatment as covariants . There was no significant association between EPS occurrence and polymorphisms in the P14416 and P35462 genes . CONCLUSIONS : An association was observed between polymorphisms in P28223 and P28335 genes and occurrence of acute EPS in schizophrenic patients treated with perphenazine monotherapy . Larger study populations are needed to confirm our findings . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Loss of the mu opioid receptor on different genetic backgrounds leads to increased bromodeoxyuridine labeling in the dentate gyrus only after repeated injection . The endogenous opioid system is involved in various physiological processes , including neurogenesis in the dentate gyrus ( DG ) of the hippocampus . In the current study , we investigated the role of the mu opioid receptor ( P35372 ) on DG neurogenesis and measured glucocorticoid levels following several injection paradigms to supplement the neurogenesis experiments . P35372 knockout ( KO ) mice on C57BL/6 and 129S6 backgrounds were injected with bromodeoxyuridine ( BrdU ) using either a single injection or two different repeated injection protocols and then sacrificed at different time points . The total number of BrdU and proliferating cell nuclear antigen ( P12004 ) positive cells in the DG is significantly increased in P35372 KO mice compared with wild type ( WT ) on both strains after repeated injection , but not after a single injection . Plasma corticosterone ( O00230 ) levels increased similarly in P35372 KO and WT mice following both single and repeated injection , indicating that the stress response is activated following any injection protocol , but that the mechanism responsible for the increase in BrdU labeling in P35372 KO mice is O00230 -level independent . Finally , WT 129S6 mice , independent of genotype , showed higher levels of plasma O00230 compared with WT C57BL/6 mice in both noninjected controls and following injection at two separate time points ; these levels were inversely correlated with low numbers of BrdU cells in the DG in 129S6 mice compared with C57BL/6 mice . In summary , these data demonstrate that loss of P35372 increases BrdU labeling in the DG independent of O00230 levels , but only following a repeated injection , illustrating the capability of injection paradigms to influence cell-proliferative responses in a genotype-dependent manner .

MH_train_1012

MH_train_1012

MH_train_1012

interacts_with DB00898?

multiple_choice

Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . New findings on the genetic influences on alcohol use and dependence . PURPOSE OF REVIEW : DB00898 dependence is a complex disorder with a well documented highly hereditary nature . This article reviews the recent advances in our understanding of the direct and indirect genetic influences on alcohol use and dependence . RECENT FINDINGS : Recent findings can be summarized as follows : ( a ) twin studies have defined and estimated the risks of general and specific alcohol-related vulnerabilities . ( b ) Linkage studies have provided largely inconsistent findings , though several chromosomal regions have been implicated . ( c ) Quantitative trait loci analyses in animals have identified that the Mpdz gene predisposes to alcohol dependence and withdrawal . ( d ) Examination of family-based samples has identified several genes including P47869 and P08172 thought to be associated with alcohol dependence . SUMMARY : Despite great advances in understanding of genetic vulnerability in alcohol use disorders , only two gene complexes , DB00067 and P05091 , have been identified as having defined effects on alcohol use and liability to dependence in humans . New genes associated with increased risks for the disorder will certainly be added to this list in the near future . Neurobiological analyses of the effects of these genes will surely contribute to further understanding of the cause of alcohol dependence and the interindividual differences in risks . Release of mediators of systemic inflammatory response syndrome in the course of a severe delayed hemolytic transfusion reaction caused by anti-D . BACKGROUND : In vitro studies suggest that mediators of systemic inflammatory response syndrome are generated in the course of hemolytic transfusion reactions . Evidence for the in vivo significance of these findings is given by the present clinical and laboratory analysis of a severe delayed hemolytic transfusion reaction ( P10275 ) . CASE REPORT : A 67-year-old patient ( blood group O , D-negative ) with a negative pretransfusion antibody screen received a massive transfusion because of arterial bleeding ( Day 1 ) . The transfusion of group O , D-positive red cell concentrates was unavoidable because of limited supplies . At Day 10 , the patient developed a P10275 with symptoms of septic-toxic syndrome and signs of hemolysis ; he received an exchange transfusion . Serologic markers , as well as proinflammatory and anti-inflammatory mediators , were monitored at the onset of the P10275 and during the exchange transfusion . RESULTS : At Day 10 , the direct antiglobulin test was positive ; anti-D was present , most likely as the result of an anamnestic immune response . Interleukin ( IL ) -1 was not detectable ; all other mediators monitored were elevated : IL-1 receptor antagonist , tumor necrosis factor , P05231 , P10145 , P22301 , neopterin , elastase , C3a-desArg , P02741 , and fibrinogen . Most of the values declined during the exchange transfusion , which was followed by an improvement of the clinical presentation . CONCLUSIONS : Mediators of systemic inflammatory response syndrome were released in the course of a P10275 caused by anti-D . Severe clinical symptoms could be treated successfully by exchange transfusion . P12821 inhibitors could be therapeutic for antisocial personality disorder . Antisocial personality traits are an important topic for research . The societal cost of these behaviors encourages efforts at a better understanding of central nervous system causes . Catecholamine genes are being studied to facilitate this understanding , and some tentative findings are being reached about several of these genes . It seems that many genes play a role to produce antisocial behaviors so complexity of elucidating each gene is obvious . One conclusion that could be drawn from the current research findings is that DA2 like receptors ( P14416 , P35462 , P21917 ) with alleles that decrease neurotransmission are facilitatory of antisocial behaviors . DA2 like receptors cause neuronal firing to inhibit many peripheral functions through adenylyl cyclase inhibition . When these receptors are less active by genetically decreased density , lower affinity , or by low dopamine levels as final common pathways then inhibition is released and a state of disinhibition can be said to describe this state . Peripheral metabolism is increased and behavioral activation is noted . P00797 is disinhibited in this setting thus allowing sympathetic nervous system activation . The fight or flight behaviors thus produced , in the extreme , would be the setting of antisocial behavior . Research validates this hypothesis . Understanding this final common pathway toward antisocial behavior should lead to better treatment for individuals with this pattern of behavior before they have caused harm to themselves and others . P12821 inhibitors are well tolerated drugs used in the treatment of hypertension and heart failure and would also treat antisocial behavior disorders . DB00898 -related expectancies are associated with the D2 dopamine receptor and GABAA receptor beta3 subunit genes . Molecular genetic research has identified promising markers of alcohol dependence , including alleles of the D2 dopamine receptor ( P14416 ) and the GABAA receptor beta3 subunit ( P28472 ) genes . Whether such genetic risk manifests itself in stronger alcohol-related outcome expectancies , or in difficulty resisting alcohol , is unknown . In the present study , A1+ ( A1A1 and A1A2 genotypes ) and A1- ( A2A2 genotype ) alleles of the P14416 and P55008 + ( G1G1 and P55008 non- P55008 genotypes ) and P55008 - ( non- P55008 non- P55008 genotype ) alleles of the P28472 gene were determined in a group of 56 medically ill patients diagnosed with alcohol dependence . Mood-related alcohol expectancy ( AE ) and drinking refusal self-efficacy ( DRSE ) were assessed using the Drinking Expectancy Profile ( Manual for the Drinking Expectancy Profile , Behaviour Research and Therapy Centre , Brisbane , 1996 ) . Patients with the P14416 A1+ allele , compared with those with the P14416 A1- allele , reported significantly lower DRSE in situations of social pressure . Similarly , lower DRSE was reported under social pressure by patients with the P28472 P55008 + allele when compared to those with the P28472 P55008 - alleles . Patients with the P28472 P55008 + allele also revealed reduced DRSE in situations characterized by negative affect than those with the P28472 P55008 - alleles . Patients carrying the P28472 P55008 + allele showed stronger AE relating to negative affective change ( for example , increased depression ) than their P28472 P55008 - counterparts . Biological influence in the development of some classes of cognitions is hypothesized . The clinical implications , particularly with regard to patient-treatment matching and the development of an integrated psychological and pharmacogenetic approach , are discussed . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Association of Q05940 gene polymorphisms with alcohol dependence . DB00898 -related diseases cause significant harm in the western world . Up to 65 % of the phenotypic variance is genetically determined . Few candidate genes have been identified , comprising P08319 , P05091 , P21964 , P34998 , Q01959 ( Q01959 ) , P47869 and P21397 . While abnormalities in the dopaminergic mesolimbic reward system are considered important mediators of alcoholism , studies analyzing variants of dopamine receptors showed conflicting results . Other modulators of the reward system are synaptosomal genes . Among candidate genes , polygenic variants of the Vesicular Monamine Transporter 2 ( Q05940 ) gene locus associated with alterations of drinking behavior were published . These variants comprise single nucleotide polymorphisms ( SNPs ) within the promoter region and the open reading frame . In this study , we confirm the association of Q05940 SNP rs363387 ( allelic association : p = 0.015 ) with alcohol dependence . This SNP defines several haplotypes including up to four SNPs ( minimal p = 0.0045 ) . In addition , numeric effects in the subgroups of males and patients with positive family history were found . We suggest that several rs363387 T-allele containing haplotypes increase the risk of alcohol dependence ( OR 1.53 ) , whereas G-allele containing haplotypes confer protection against alcohol dependence . Taken together , there is supporting evidence for a contribution of Q05940 gene variants to phenotypes of alcohol dependence . The P38936 codon 31*C- and P14416 codon 313*T-related genotypes/alleles , but not P18887 codon 399 , hOGG1 codon 326 , and P21728 -48 polymorphisms , are correlated with the presence of leiomyoma . OBJECTIVE : To investigate whether the gene polymorphisms for P38936 , X-ray repair cross-complementing group 1 ( P18887 ) , human 8-oxoguanine glycosylase 1 ( hOGG1 ) , and dopamine D1 and D2 receptors ( P21728 , -2 ) are associated with leiomyoma susceptibility . DESIGN : Prospective study . SETTING : Departments of gynecology and genetics in a medical center . PATIENT(S) : Women were divided into two groups : leiomyoma ( n = 120 ) and nonleiomyoma ( n = 112 ) . INTERVENTION(S) : The P38936 codon 31 , P18887 codon 399 , hOGG1 codon 326 , P21728 -48 , and P14416 codon 313 polymorphisms were genotyped by polymerase chain reaction with restriction enzyme digestions ( Blp I , MspI , Fnu4HI , Dde I , and NcoI , respectively ) . MAIN OUTCOME MEASURE(S) : Genotypes and allelic frequencies . RESULT(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyomas . The proportions of P38936 (*)CC/CA/AA and P14416 (*)CC/CT/TT in both groups were 27.5/68.3/4.2 % and 12.5/51.7/35.8 % ( leiomyoma ) ; and 14.3/51.8/33.9 % and 33.9/40.2/25.9 % ( nonleiomyoma ) . P18887 , hOGG1 , and P21728 were not correlated with the presence of leiomyomas . P18887 (*)GG/GA/AA , hOGG1(*)TT/TA/AA , and P21728 (*)GG/GA/AA were 54.2/37.5/8.3 % , 36.7/44.2/19.1 % , and 3.3/25.8/70.8 % ( leiomyoma ) ; and 48.2/47.3/4.5 % , 43.6/41/15.4 % , and 3.6/25/71.4 % ( nonleiomyoma ) . CONCLUSION(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyoma . P18887 , hOGG1 , and P21728 were not correlated with leiomyoma development . Multiple cholinergic nicotinic receptor genes affect nicotine dependence risk in African and European Americans . Several independent studies show that the chromosome 15q25.1 region , which contains the P30532 - P32297 - P30926 gene cluster , harbors variants strongly associated with nicotine dependence , other smoking behaviors , lung cancer and chronic obstructive pulmonary disease . We investigated whether variants in other cholinergic nicotinic receptor subunit ( CHRN ) genes affect the risk of nicotine dependence in a new sample of African Americans ( AAs ) ( N = 710 ) . We also analyzed this AA sample together with a European American ( EA ) sample ( N = 2062 , 1608 of which have been previously studied ) , allowing for differing effects in the two populations . Cases are current nicotine-dependent smokers and controls are non-dependent smokers . Variants in or near Q07001 - P07510 , P36544 and Q9GZZ6 show modest association with nicotine dependence risk in the AA sample . In addition , P43681 , Q05901 - Q15825 and P11230 show association in at least one population . P07510 and P43681 harbor single nucleotide polymorphisms ( SNPs ) that have opposite directions of effect in the two populations . In each of the population samples , these loci substantially increase the trait variation explained , although no loci meet Bonferroni-corrected significance in the AA sample alone . The trait variation explained by three key associated SNPs in P30532 - P32297 - P30926 is 1.9 % in EAs and also 1.9 % in AAs ; this increases to 4.5 % in EAs and 7.3 % in AAs when we add six variants representing associations at other CHRN genes . Multiple nicotinic receptor subunit genes outside chromosome 15q25 are likely to be important in the biological processes and development of nicotine dependence , and some of these risks may be shared across diverse populations . DB09073 ( PD 0332991 ) : targeting the cell cycle machinery in breast cancer . INTRODUCTION : The cyclin D-cyclin-dependent kinases 4 and 6 ( P11802 /6 ) -retinoblastoma ( P06400 ) pathway , governing the cell cycle restriction point , is frequently altered in breast cancer and is a potentially relevant target for anticancer therapy . DB09073 ( PD 0332991 ) , a potent and selective inhibitor of P11802 and Q00534 , inhibits proliferation of several P06400 -positive cancer cell lines and xenograft models . AREAS COVERED : The basic features and abnormalities of the cell cycle in breast cancer are described , along with their involvement in estrogen signaling and endocrine resistance . The pharmacological features of palbociclib , its activity in preclinical models of breast cancer and the potential determinants of response are then illustrated , and its clinical development in breast cancer described . A literature search on the topic was conducted through PubMed and the proceedings of the main cancer congresses of recent years . EXPERT OPINION : The combination of palbociclib with endocrine agents is a very promising treatment and Phase III clinical trials are ongoing to confirm its efficacy . Further , potentially useful combinations are those with drugs targeting mitogenic signaling pathways , such as P04626 - and PI3K-inhibitors . Combination with chemotherapy seems more problematic , as antagonism has been reported in preclinical models . The identification of predictive factors , already explored in preclinical studies , must be further refined and validated in clinical trials . [ Genetic aspects of occupational chronic obstructive lung disease under exposure to various risk factors ] . The article deals with data on association of SNP rs1828591 of Q96QV1 gene with COLD development under exposure to dust and chemical factors . SNP rs1800470 of TGFbeta1 gene is associated with occupational COLD under exposure to dust and did not show connection with COLD under exposure to chemical aerosols . No association was seen between SNP rs4129267 of IL-6R gene and SNP rs1051730 of P32297 gene with occupational COLD under exposure to the studied factors . SNP rs1828591 of Q96QV1 gene is associated with occupational COLD development under exposure to dust and chemical factors . Study of association of genotype and phenotypic features of COLD revealed the following trends : " dust " COLD patients with genotype AA SNP rs1800470 of TGFbeta1 gene show lower level of P02741 and P01375 , if compared with other genotypes . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Expression profiling of gastric adenocarcinoma using cDNA array . To investigate the expression profile of gastric adenocarcinoma , cDNA array experiments were performed using Atlas Human Cancer 1.2 K Array ( Clontech Laboratories , Palo Alto , CA ) on nine xenografted and two primary gastric cancer samples . The expression of the tumor samples was compared to that of two normal gastric epithelial tissues . The expression pattern of the primary tumors was similar to that of xenografted tumors . The up-regulated genes had expression ratios ranging from 2.5 to 16 , whereas the down-regulated genes had a range from -2.5 to -16 . No variation in gene expression was detected in the analysis of the xenografted tumors versus the primary tumors , indicating that the xenografts represented the primary tumors well . Thirty-eight genes showed altered gene expression in 5 or more samples ( > 45 % ) . Thirty-one genes were up-regulated and seven genes were down-regulated . The most abundantly up-regulated genes ( ratio > 5 ) included genes such as P26447 , P11802 , P50281 and beta catenin . The GIF was markedly down-regulated ( ratio < -10 ) . To confirm our findings , six genes ( three up- and three down-regulated ) were selected for semi-quantitative RT-PCR analysis . The RT-PCR results were consistent with the array findings . Our approach revealed that several genes are abnormally expressed in gastric cancer and found that genes known to interact in several common molecular pathway(s) were consistently altered . DB00563 gamma-hydroxamate derivatives as potential dual target antitumor drugs . A series of new aminopteroyl-based hydroxamate derivatives were synthesized and tested in vitro in cell culture models as potential dual target drugs . These compounds were designed to target two families of enzymes , matrix metalloproteinases ( MMP ) and a folate enzyme , dihydrofolate reductase ( P00374 ) . These enzymes are the components of two unrelated cellular pathways and they are often over-expressed in metastasizing tumors . In addition to the synthesis and full structural characterization of the hybrid molecules , we describe their inhibitory activities against a series of MMPs ( P08253 , P09237 , P14780 , P50281 ) and P00374 , as well as their antiproliferative activity in three cancer cell lines . The new hydroxamate derivatives of MTX proved to be effective inhibitors of MMPs and P00374 in the micromolar and nanomolar range , respectively . Furthermore , they showed strong antiproliferative activity against A549 cells ( non-small cell lung carcinoma ) , and PPC-1 and Tsu-Pr1 prostate cancer cell lines . Therefore , based on the present results , these bi-functional drugs may be good candidates to target specific tumors in animal models due to potential combined effects on two pathways crucial for tumor development . DB00898 blocks leukocyte recruitment and endothelial cell activation in vivo and in vitro . Immune system impairment and increased susceptibility to infection among alcohol abusers is a significant but not well-understood problem . We hypothesized that acute ethanol administration would inhibit leukocyte recruitment and endothelial cell activation during inflammation and infection . Using LPS and carrageenan air pouch models in mice , we found that physiological concentrations of ethanol ( 1-5 g/kg ) significantly blocked leukocyte recruitment ( 50-90 % ) . Because endothelial cell activation and immune cell-endothelial cell interactions are critical regulators of leukocyte recruitment , we analyzed the effect of acute ethanol exposure on endothelial cell activation in vivo using the localized Shwartzman reaction model . In this model , ethanol markedly suppressed leukocyte accumulation and endothelial cell adhesion molecule expression in a dose-dependent manner . Finally , we examined the direct effects of ethanol on endothelial cell activation and leukocyte-endothelial cell interactions in vitro . DB00898 , at concentrations within the range found in human blood after acute exposure and below the levels that induce cytotoxicity ( 0.1-0.5 % ) , did not induce endothelial cell activation , but significantly inhibited P01375 -mediated endothelial cell activation , as measured by adhesion molecule ( P16581 , P05362 , P19320 ) expression and chemokine ( P10145 , P13500 , RANTES ) production and leukocyte adhesion in vitro . Studies exploring the potential mechanism by which ethanol suppresses endothelial cell activation revealed that ethanol blocked NF-kappaB nuclear entry in an P25963 -dependent manner . These findings support the hypothesis that acute ethanol overexposure may increase the risk of infection and inhibit the host inflammatory response , in part , by blocking endothelial cell activation and subsequent immune cell-endothelial cell interactions required for efficient immune cell recruitment . Most reported genetic associations with general intelligence are probably false positives . General intelligence ( g ) and virtually all other behavioral traits are heritable . Associations between g and specific single-nucleotide polymorphisms ( SNPs ) in several candidate genes involved in brain function have been reported . We sought to replicate published associations between g and 12 specific genetic variants ( in the genes Q96EV8 , P07339 , P14416 , Q8NFD2 , P08172 , P51649 , P21964 , P23560 , P43681 , Q9NRI5 , P02649 , and P60880 ) using data sets from three independent , well-characterized longitudinal studies with samples of 5,571 , 1,759 , and 2,441 individuals . Of 32 independent tests across all three data sets , only 1 was nominally significant . By contrast , power analyses showed that we should have expected 10 to 15 significant associations , given reasonable assumptions for genotype effect sizes . For positive controls , we confirmed accepted genetic associations for Alzheimer 's disease and body mass index , and we used SNP-based calculations of genetic relatedness to replicate previous estimates that about half of the variance in g is accounted for by common genetic variation among individuals . We conclude that the molecular genetics of psychology and social science requires approaches that go beyond the examination of candidate genes . Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced P28906 + cell-derived human dendritic cells . The efficient genetic modification of P28906 + cell-derived dendritic cells ( DC ) will provide a significant advancement towards the development of immunotherapy protocols for cancer , autoimmune disorders and infectious diseases . Recent reports have described the transduction of P28906 + cells via retrovirus- and lentivirus-based gene transfer vectors and subsequent differentiation into functional DC . Since there is significant apprehension regarding the clinical uses of HIV-based vectors , in this report , we compare a murine leukemia virus ( MLV ) - and a human immunodeficiency virus ( HIV ) -based bicistronic vector for gene transfer into human P28906 + cells and subsequent differentiation into mature DC . Each vector expressed both EGFP and the dominant selectable marker P00374 (L22Y) allowing for the enrichment of marked cells in the presence of the antifolate drug trimetrexate ( TMTX ) . Both MLV-based and HIV-based vectors efficiently transduced cytokine mobilized human peripheral blood P28906 + cells . However , in vitro expansion and differentiation in the presence of GM- P04141 , P01375 , Flt-3L , P21583 and P05112 resulted in a reduction in the percentage of DC expressing the transgene . Selection with TMTX during differentiation increased the percentage of marked DC , resulting in up to 79 % ( MLV vector ) and up to 94 % ( lentivirus-vector ) transduced cells expressing EGFP without loss of DC phenotype . Thus , MLV-based vectors and in vitro selection of transduced human DC show great promise for immunotherapy protocols . The novel DB01221 receptor antagonist , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid , is a gating modifier in cultured mouse cortical neurons . Neu2000 [ P04626 , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid ] , a derivative of sulfasalazine , attenuates DB01221 -induced neuronal toxicity . Here we investigated the effects of P04626 on the DB01221 receptor ( NMDAR ) using whole-cell patch clamp technique to determine the molecular mechanisms underlying its neuroprotective role . P04626 reversibly suppressed DB01221 responses in an uncompetitive manner with fast binding kinetics . Its inhibition of NMDAR activity depended on both the concentration and the use of agonist but not on the membrane potential . P04626 accelerated DB01221 desensitization without affecting the binding affinity of NMDAR for its agonists and stabilized the closed state of NMDAR . Therefore , P04626 should effectively alleviate disorders that are a result of glutamate excitoxicity with fewer side effects because it is a low-affinity gating modifier that antagonizes NMDAR in an uncompetitive manner . Moreover , in the presence of ifenprodil ( an Q13224 antagonist ) but not DB00238 -AAM077 [ ( R ) - [ ( S ) -1-(4-bromo-phenyl)-ethylamino ] -(2,3-dioxo-1,2,3,4-tetrahydro-quinoxalin-5-yl)-methyl ] -phosphonic acid , an Q12879 antagonist ] , the extent of P04626 block was decreased , suggesting that P04626 is an Q13224 -specific antagonist . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . Long-term exposure to methotrexate induces immunophenotypic changes , decreased methotrexate uptake and increased dihydrofolate gene copy number in Jurkat T cells . DB00563 ( MTX ) treatment of rheumatoid arthritis may require increasing doses to maintain clinical efficacy . An overall plateau of clinical response is reached after only six months of treatment . To study the immunologic , biochemical and genetic effects of MTX on T cells , the Jurkat T cell line was made MTX-resistant by serial addition of methotrexate sodium into culture medium . Cells proliferated and divided successfully in MTX concentrations ranging to 15 microM . MTX resistance of Jurkat T cells in vitro was accompanied by significantly ( P < 0.05 ) decreased expression of P06729 , CD3 , P01730 , P10747 , and Q07108 , P60568 production , and MTX uptake assessed by cell association or disassociation of 3[H]-MTX or fluoresceinated MTX ( FMTX ) , respectively . In addition , there was P00374 gene amplification and increased levels of P00374 in all resistant cell lines . Both permanent and transient phenotypic changes developed in resultant cell lines exposed to increasing concentrations of MTX in vitro . Expression of P01730 and CD25 and sensitivity to MTX returned to near-parental levels after removal of MTX from culture medium , whereas expression of P27487 and MTX uptake were significantly increased . Expression of P06729 , CD3 , Q07108 and P60568 production as well as the P00374 levels did not return to the parental phenotype after removal from MTX . We conclude that MTX-cultured cells express depressed levels of cell-surface markers vital for T cell function and activation . The return of enhancement of these cell-surface markers critical to T cell activation suggests a possible mechanism for the severe flares experienced by rheumatoid arthritis patients when drug treatment is discontinued . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . DB00898 dose-dependently elicits opposing regulatory effects on hippocampal AMPA receptor P42262 subunits through a zeta inhibitory peptide-sensitive kinase in adolescent and adult Sprague-Dawley rats . AMPA receptor P42262 subunits are strongly implicated in cognition , and prior work suggests that these subunits may be regulated by atypical protein kinase C ( aPKC ) isoforms . The present study assessed whether hippocampal and cortical AMPA receptor P42262 subunit regulation may be an underlying factor in known age-related differences to cognitive-impairing doses of ethanol , and if aPKC isoforms modulate such responses . Hippocampal AMPA receptor P42262 subunit , protein kinase Mζ ( PKMζ ) , and PKCι/λ expression were elevated during adolescence compared to adults . 1 h following a low-dose ( 1.0-g/kg ) ethanol exposure , hippocampal AMPA receptor P42262 subunit serine 880 phosphorylation was decreased in adolescents , but was increased in adults . Age-dependent changes in P42262 subunit phosphorylation were paralleled by alterations in aPKC isoforms , and zeta inhibitory peptide ( Q8N5A5 ) administration prevented ethanol-induced increases in both in adults . DB00898 -induced changes in P42262 subunit phosphorylation were associated with delayed regulation in synaptosomal P42262 subunit expression 24 h later . A higher ethanol dose ( 3.5-g/kg ) failed to elicit changes in most measures in the hippocampus at either age . Similar to the hippocampus , analysis of cerebral cortical tissue also revealed age-related declines . However , no demonstrable effects were found following a low-dose ethanol exposure at either age . High-dose ethanol exposure reduced adolescent P42262 subunit phosphorylation and aPKC isoform expression that were again accompanied by delayed reductions in synaptosomal P42262 subunit expression . Together , these results suggest that P42262 -containing AMPA receptor modulation by aPKC isoforms is age- , region- and dose-dependently regulated , and may potentially be involved in developmentally regulated ethanol-induced cognitive impairment and other ethanol behaviors . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Genetics of alcoholism . DB00898 use and alcohol use disorders are substantially heritable . Variants in genes coding for alcohol metabolic enzymes have long been known to influence consumption . More recent studies in family-based samples have implicated P47869 , nicotinic receptor genes such as Q05901 , and a number of other specific single genes as associated with alcohol use disorders . The growing use of genetic analyses , in particular studies using polygenic risk scores ; neurobiologic pathways ; and methods for quantifying gene × gene and gene × environment interactions have also contributed to an evolving understanding of the genetic architecture of alcohol use disorders . Additionally , the study of behavioral traits associated with alcohol dependence such as impulsivity and sensation seeking , and the influences of demographic factors ( i.e. , sex and ethnicity ) have significantly enhanced the genetics of alcoholism literature . This article provides a brief overview of the current topically relevant findings in the field to date and includes areas of research still requiring attention . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . Comparison of three GPCR structural templates for modeling of the Q9H244 nucleotide receptor . The P2Y(12) receptor ( P2Y(12)R ) is an ADP-activated G protein-coupled receptor ( GPCR ) that is an important target for antithrombotic drugs . Three homology models of P2Y(12)R were compared , based on different GPCR structural templates : bovine rhodopsin ( bRHO ) , human A(2A) adenosine receptor ( A(2A)AR ) , and human P61073 ( P61073 ) . By criteria of sequence analysis ( 25.6 % identity in transmembrane region ) , deviation from helicity in the second transmembrane helix ( TM2 ) , docked poses of ligands highlighting the role of key residues , accessibility of a conserved disulfide bridge that is reactive toward irreversibly-binding antagonists , and the presence of a shared disulfide bridge between the third extracellular loop ( EL3 ) and the N-terminus , the P61073 -based model appeared to be the most consistent with known characteristics of P2Y(12)R . The docked poses of agonist 2MeSADP and charged anthraquinone antagonist PSB-0739 in the binding pocket of P2Y(12)R-CXC agree with previously published site-directed mutagenesis studies of Arg256 and Lys280 . A sulfonate at position 2 of the anthraquinone core created a strong interaction with the Lys174(EL2) side chain . The docking poses of the irreversibly-binding , active metabolite ( existing as two diastereoisomers in vivo ) of the clinically utilized antagonist DB00758 were compared . The free thiol group of the 4S diastereoisomer , but not the 4R isomer , was found in close proximity ( ~4.7 Å ) to the sulfur atom of a disulfide bridge involving Cys175 , suggesting greater activity in covalent binding . Therefore , ligand docking to the P61073 -based model of the P2Y(12)R predicted poses of both reversibly and irreversibly-binding small molecules , consistent with observed pharmacology and mutagenesis studies . Bacterial translocation in cirrhotic rats stimulates P29474 -derived NO production and impairs mesenteric vascular contractility . DB00435 ( NO ) has been implicated in the arterial vasodilation and associated vascular hyporesponsiveness to vasoconstrictors observed in liver cirrhosis . Bacteria , potent activators of NO and P01375 synthesis , are found in the mesenteric lymph nodes ( MLNs ) of ascitic cirrhotic rats . Here , we investigated the impact of bacterial translocation ( BT ) to MLNs on P01375 production , vascular NO release , and contractility in the mesenteric vasculature of ascitic cirrhotic rats . Vascular response to the alpha-adrenoagonist methoxamine , which is diminished in the superior mesenteric arterial beds of cirrhotic rats , is further blunted in the presence of BT . BT promoted vascular NO release in cirrhotic rats , an effect that depended on pressure-induced shear stress and was blocked by the NO inhibitor N(omega)-nitro-L-arginine . Removing the endothelium had the same effect . Endothelial NO synthase ( P29474 ) , but not the inducible isoform ( P35228 ) , was present in mesenteric vasculature of cirrhotic rats with and without BT , and its expression was enhanced compared with controls . P01375 was induced in MLNs by BT and accumulated in parallel in the serum . This P01375 production was associated with elevated levels of tetrahydrobiopterin ( BH(4) ) , a P01375 -stimulated cofactor and enhancer of P29474 -derived NO biosynthesis and NOS activity in mesenteric vasculature . These findings establish a link between BT to MLNs and increased P01375 production and elevated BH(4) levels enhancing P29474 -derived NO overproduction , further impairing contractility in the cirrhotic mesenteric vasculature . Predicting the effect of naltrexone and acamprosate in alcohol-dependent patients using genetic indicators . DB00659 and naltrexone are effective medications in the treatment of alcoholism . However , effect sizes are modest . Pharmacogenomics may improve patient-treatment-matching and effect sizes . It is hypothesized that naltrexone exerts its effect through genetic characteristics associated with the dopaminergic/opioidergic positive reinforcement system , whereas acamprosate works through the glutamatergic/GABAergic negative reinforcement system . DB00898 -dependent subjects were randomly assigned to either acamprosate or naltrexone . Subjects participated in a cue-exposure experiment at the day before and at the last day of medication . Reductions in cue-induced craving and physiological cue reactivity were measured . Differential effects of naltrexone and acamprosate on these outcomes were tested for different polymorphisms of the opioid , dopamine , glutamate and GABA-receptors . Significant matching effects were found for polymorphisms at the P14416 , Q16445 and P47870 gene . In addition , a trend was found for the P35372 polymorphism . This provides evidence for the matching potential of genotypes . It is expected that more effective treatments can be offered when genetic information is used in patient-treatment-matching . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . Effects of exogenous spermidine on photosynthetic capacity and expression of Calvin cycle genes in salt-stressed cucumber seedlings . We investigated the effects of exogenous spermidine ( Spd ) on growth , photosynthesis and expression of the Calvin cycle-related genes in cucumber seedlings ( Cucumis sativus L. ) exposed to NaCl stress . Salt stress reduced net photosynthetic rates ( PN ) , actual photochemical efficiency of PSII ( ΦPSII ) and inhibited plant growth . Application of exogenous Spd to salinized nutrient solution alleviated salinity-induced the inhibition of plant growth , together with an increase in PN and ΦPSII . Salinity markedly reduced the maximum carboxylase activity of ribulose-1,5-bisphosphate carboxylase/oxygenase ( Vcmax ) , the maximal velocity of RuBP regeneration ( Jmax ) , triose-phosphate utilization capacity ( TPU ) and carboxylation efficiency ( CE ) . Spd alleviated the negative effects on CO2 assimilation induced by salt stress . Moreover , Spd significantly increased the activities and contents of ribulose-1,5-bisphosphate carboxylase/oxygenase ( Rubisco ) and fructose-1,6-biphosphate aldolase ( P33897 ; aldolase ) in the salt-stressed cucumber leaves . On the other hand , salinity up-regulated the transcriptional levels of ribulose-1,5-bisphosphate ( RCA ) , glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) and phosphoribrokinase ( Q9H4B4 ) and down-regulated the transcriptional levels of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit ( RbcL ) , ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit ( RbcS ) , P33897 , triose-3-phosphate isomerase ( P60174 ) , fructose-1,6-bisphosphate phosphatase ( FBPase ) and 3-phosphoglyceric acid kinase ( PGK ) . However , Spd application to salt-stressed plant roots counteracted salinity-induced mRNA expression changes in most of the above-mentioned genes . These results suggest that Spd could improve photosynthetic capacity through regulating gene expression and activity of key enzymes for CO2 fixation , thus confers tolerance to salinity on cucumber plants . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Selective increases of AMPA , DB01221 , and kainate receptor subunit mRNAs in the hippocampus and orbitofrontal cortex but not in prefrontal cortex of human alcoholics . Glutamate is the main excitatory transmitter in the human brain . Drugs that affect the glutamatergic signaling will alter neuronal excitability . DB00898 inhibits glutamate receptors . We examined the expression level of glutamate receptor subunit mRNAs in human post-mortem samples from alcoholics and compared the results to brain samples from control subjects . RNA from hippocampal dentate gyrus ( HP-DG ) , orbitofrontal cortex ( OFC ) , and dorso-lateral prefrontal cortex ( DL- P27918 ) samples from 21 controls and 19 individuals with chronic alcohol dependence were included in the study . Total RNA was assayed using quantitative RT-PCR . Out of the 16 glutamate receptor subunits , mRNAs encoding two AMPA [ 2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid ] receptor subunits P42262 and P42263 ; three kainate receptor subunits Q13002 , Q13003 and Q16478 and five DB01221 ( N-methyl-D-aspartate ) receptor subunits Q05586 , Q12879 , Q14957 , O15399 , and Q8TCU5 were significantly increased in the HP-DG region in alcoholics . In the OFC , mRNA encoding the DB01221 receptor subunit Q8TCU5 was increased , whereas in the DL- P27918 , no differences in mRNA levels were observed . Our laboratory has previously shown that the expression of genes encoding inhibitory GABA-A receptors is altered in the HP-DG and OFC of alcoholics ( Jin et al. , 2011 ) . Whether the changes in one neurotransmitter system drives changes in the other or if they change independently is currently not known . The results demonstrate that excessive long-term alcohol consumption is associated with altered expression of genes encoding glutamate receptors in a brain region-specific manner . It is an intriguing possibility that genetic predisposition to alcoholism may contribute to these gene expression changes . Selective measurement of white matter and gray matter diffusion trace values in normal human brain . The trace of the diffusion tensor ( or simply the trace ) is diagnostically valuable for detecting acute ischemic lesions . A number of studies indicate that the trace of human gray matter ( GM ) and white matter ( WM ) are quite similar . This is somewhat surprising considering the different cellular environments of GM and WM . It is possible that partial volume averaging ( P32926 ) effects between GM and WM , inherent in many of the ultrafast imaging sequences used for diffusion measurements , are responsible for this observation . In order to minimize P32926 effects , the trace values of GM and WM have been selectively measured by implementing double inversion recovery ( P30518 ) echo planar imaging ( P10646 ) pulse sequences . Results on six normal volunteers indicate that the trace values of WM and GM are not statistically different . Differential increases in catecholamine metabolizing enzymes in amyotrophic lateral sclerosis . The activity of three catecholamine-metabolizing enzymes , monoamine oxidase type A and type B ( P21397 and P27338 ) as well as catechol-O-methyltransferase ( P21964 ) , were estimated in homogenates of human spinal cord using radiometric assays . The enzyme activities were determined in postmortem spinal cord tissue from controls and cases with amyotrophic lateral sclerosis ( P35858 ) . The activity of P21397 was below the limit of detectability in both controls and P35858 cases . The activities of P27338 and P21964 were evenly distributed at the various spinal levels . The P27338 activity was substantially elevated in P35858 spinal homogenates , whereas only a slight , but not statistically significant , increase in P21964 activity was observed . A significant correlation between P21964 and P27338 activities was observed for controls . However , this covariation was not apparent for the P35858 cases . These results suggest that the two enzyme proteins are regulated by more complex mechanisms in the spinal cord in amyotrophic lateral sclerosis than simple general increases caused by elevated astroglial cell numbers . In addition , the P21397 , P27338 , and P21964 activities were estimated in spinal cords from rats treated with the selective P27338 inhibitor L-deprenyl , a drug with putative neuroprotective effects in neurodegenerative disorders . After 3 weeks of L-deprenyl treatment ( 0.25 mg/kg/day , sc ) , the spinal P21397 and P27338 activities were decreased by 50 and 80 % , respectively . In contrast , the P21964 activity was not altered by L-deprenyl administration . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . Downstream effects of striatal-enriched protein tyrosine phosphatase reduction on RNA expression in vivo and in vitro . Striatal-enriched protein tyrosine phosphatase ( P54829 ) is a brain-specific tyrosine phosphatase that has been shown to de-phosphorylate several key neuronal signaling proteins , including kinases ( extracellular signal-regulated kinase ( P27361 /2 ) , P06241 , Q14289 ) and glutamate receptor subunits ( N-methyl-d-aspartate receptor subtype 2B ( Q13224 ) , glutamate receptor 2 ( P42262 ) ) . Step knock-out mice have increased phosphorylation of these substrates in the brain , with potential functional consequences in synaptic plasticity and cognitive tasks . It is therefore of interest to identify the molecular pathways and downstream transcriptional targets that are impacted by Step knockdown . In the present study , striatal RNA samples from Step wild-type , knock-out and heterozygous mice were hybridized to Affymetrix microarray chips and evaluated for transcriptional changes between genotypes . Pathway analysis highlighted Erk signaling and multiple pathways related to neurotrophin signaling , neuronal development and synaptic transmission . Potential genes of interest identified by microarray were confirmed by quantitative real-time polymerase chain reaction ( qRT-PCR ) in the cortex and hippocampus , which shared several transcriptional alterations with the striatum . In order to evaluate Step knockdown in an in vitro system , a panel of genes were evaluated using qRT-PCR in rat cortical neurons that were transduced with lentivirus expressing short hairpin RNA against Step or a non-targeting control . Our data suggest that Step has a role in the expression of immediate early genes relevant to synaptic plasticity , in both in vitro and in vivo systems . Genetic variation in the P30532 gene affects mRNA levels and is associated with risk for alcohol dependence . DB00898 dependence frequently co-occurs with cigarette smoking , another common addictive behavior . Evidence from genetic studies demonstrates that alcohol dependence and smoking cluster in families and have shared genetic vulnerability . Recently a candidate gene study in nicotine dependent cases and nondependent smoking controls reported strong associations between a missense mutation ( rs16969968 ) in exon 5 of the P30532 gene and a variant in the 3'-UTR of the P32297 gene and nicotine dependence . In this study we performed a comprehensive association analysis of the P30532 - P32297 - P30926 gene cluster in the Collaborative Study on the Genetics of Alcoholism ( COGA ) families to investigate the role of genetic variants in risk for alcohol dependence . Using the family-based association test , we observed that a different group of polymorphisms , spanning P30532 - P32297 , demonstrate association with alcohol dependence defined by Diagnostic and Statistical Manual of Mental Disorders , 4th edn ( DSM-IV ) criteria . Using logistic regression we replicated this finding in an independent case-control series from the family study of cocaine dependence . These variants show low linkage disequilibrium with the SNPs previously reported to be associated with nicotine dependence and therefore represent an independent observation . Functional studies in human brain reveal that the variants associated with alcohol dependence are also associated with altered steady-state levels of P30532 mRNA . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . An in vitro model of human acute ethanol exposure that incorporates P49682 - and P61073 -dependent recruitment of immune cells . Alcoholic liver disease ( P33897 ) is one of the commonest causes of cirrhosis and liver failure in the developed world . Hepatic inflammation is the critical stage in progression of both P33897 and non- P33897 , but it remains difficult to study the underlying mechanisms in a human system , and current animal models do not fully recapitulate human liver disease . We developed a human tissue-based system to study lymphocyte recruitment in response to ethanol challenge . Precision-cut liver slices ( PCLS ) from human livers were incubated in culture , and hepatic function was determined by albumin production , 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide assay , glucose uptake responses , and morphometric assessment . Responses of tissue and lymphocytes to ethanol exposure were determined by PCR , flow cytometry , histology , and lymphocyte infiltration assays . Human PCLS demonstrated appropriate upregulation of P05181 , ADH1α , and P00326 in response to ethanol treatment . DB00898 also induced expression of endothelial P19320 and P05362 , production of sICAM-1 and P10145 , and the chemokine receptors P49682 and P61073 on P01730 and CD8 lymphocytes . P49682 - and P61073 -dependent migration of lymphocytes into the tissue increased significantly in response to treatment with ethanol . We have demonstrated that ethanol increases chemokine receptor expression and lymphocyte recruitment into human liver tissue , suggesting that it may operate directly to promote hepatitis in P33897 . The physiological and pathophysiological responses of the PCLS to ethanol in vitro highlight the potential of this assay for dissecting the molecular mechanisms underlying human liver inflammation and as a screening tool for novel therapeutics . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Hypoxic damage to the periventricular white matter in neonatal brain : role of vascular endothelial growth factor , nitric oxide and excitotoxicity . The present study examined factors that may be involved in the development of hypoxic periventricular white matter damage in the neonatal brain . Wistar rats ( 1-day old ) were subjected to hypoxia and the periventricular white matter ( corpus callosum ) was examined for the mRNA and protein expression of hypoxia-inducible factor-1alpha ( HIF-1alpha ) , endothelial , neuronal and inducible nitric oxide synthase ( P29474 , P29475 and P35228 ) , vascular endothelial growth factor ( P15692 ) and N-methyl-D-aspartate receptor subunit 1 ( Q05586 ) between 3 h and 14 days after hypoxic exposure by real-time RT-PCR , western blotting and immunohistochemistry . Up-regulated mRNA and protein expression of HIF-1alpha , P15692 , Q05586 , P29474 , P29475 and P35228 in corpus callosum was observed in response to hypoxia . Q05586 and P35228 expression was found in the activated microglial cells , whereas P15692 was localized to astrocytes . An enzyme immunoassay showed that the P15692 concentration in corpus callosum was significantly higher up to 7 days after hypoxic exposure . NO levels , measured by colorimetric assay , were also significantly higher in hypoxic rats up to 14 days after hypoxic exposure as compared with the controls . A large number of axons undergoing degeneration were observed between 3 h and 7 days after the hypoxic exposure at electron-microscopic level . Our findings point towards the involvement of excitotoxicity , P15692 and NO in periventricular white matter damage in response to hypoxia . Estrogenic regulation of tissue factor and tissue factor pathway inhibitor in platelets . Oral estrogen treatment increases thrombotic risk . P13726 ( TF ) , tissue factor pathway inhibitor ( P10646 ) , and platelet interaction with leukocytes are important determinants of thrombogenesis . Therefore , the present study was designed to define and compare platelet TF and P10646 mRNA and adhesion protein expression in platelets derived from animals treated with different types of oral estrogens . Ovariectomized pigs were treated with 17beta-estradiol ( 2 mg/day ) , conjugated equine estrogen ( Q7L5D6 ; 0.625 mg/day ) , or raloxifene ( 60 mg/day ) for 4 wk . Compared with intact animals , ovariectomy and treatment differentially affected populations of leukocytes : neutrophils decreased whereas lymphocytes increased significantly 4 wk after ovariectomy and with 17beta-estradiol and Q7L5D6 treatments ; eosinophils increased only with 17beta-estradiol treatment . Content of TF protein increased in platelets from 17beta-estradiol- and raloxifene-treated pigs , whereas TF mRNA was detected only in platelets from 17beta-estradiol- and Q7L5D6 treated pigs . P10646 mRNA increased in platelets after ovariectomy and estrogen treatment . Only a trace of P10646 protein was detected , but a higher-molecular-mass protein was observed in all treatment groups . Expression of P25942 and P29965 increased with ovariectomy and decreased with 17beta-estradiol and Q7L5D6 treatments more than with raloxifene . The ratio of activated to basal P16109 expression decreased with ovariectomy and increased with raloxifene treatments . These results suggest that estrogenic formulations may affect individual thrombotic risk by different mechanisms that regulate TF and platelet-leukocytic interactions . These studies provide the rationale for evaluation of interactions among platelets and TF and P10646 expression on thrombin generation during estrogen treatment in humans . DB00898 increases desensitization of recombinant P48058 AMPA receptor and TARP combinations . Glutamate receptors are important target molecules of the acute effect of ethanol . We studied ethanol sensitivity of homomeric P48058 receptors expressed in human embryonic kidney 293 cells and examined whether recently discovered transmembrane alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ( AMPA ) receptor regulatory proteins ( TARPs ) affect ethanol sensitivity . Coexpression of the TARPs , stargazin , and gamma4 increased the time constant ( tau-value ) of current decay in the presence of agonist , thus slowing the onset of desensitization and increasing the steady-state current . DB00898 produced less inhibition of the peak current than the steady-state current for all types of the P48058 receptors . In addition , ethanol concentration-dependently accelerated the rate of desensitization , measured as the tau-value of fast decay of peak current . This effect was enhanced with coexpression of TARPs . The recovery from desensitization was slowed down by coexpression of gamma4 but ethanol did not affect this process in any P48058 combination . The results support the idea that increased desensitization is an important mechanism in the ethanol inhibition of AMPA receptors and indicate that coexpression of TARPs can alter this effect of ethanol . Dedifferentiated chondrosarcoma mimicking a giant cell tumor . Is this low grade dedifferentiated chondrosarcoma ? We report a very rare case of a dedifferentiated chondrosarcoma mimicking a benign giant cell tumor . A 22-year-old male was admitted to our hospital with a history of mild left wrist pain after a skiing trauma . Radiology revealed an extensive meta-epiphyseal osteolytic lesion in the distal ulna , which appeared to be a giant cell tumor . Histological examination showed a biphasic tumor comprising chondroid and non-chondroid areas with a giant cell-rich lesion resembling a conventional giant cell tumor of the bone . Immunohistochemistry showed no expression of p16(INK4a) , P17948 , P35968 ( P35968 ) , P35916 , cKIT , Q00987 or P11802 . However , high expression of the tyrosine kinases P16234 and P09619 was observed . Molecular analysis showed no amplification of the cMYC gene and no activating mutations in the cKIT ( exons 9 and 11 ) or P16234 ( exon 18 ) genes . He has been on follow-up for ten months , with no evidence of local recurrence or metastatic disease . In summary , this report highlights a very rare case of a dedifferentiated chondrosarcoma in which the dedifferentiated component of the tumor bears histologic resemblance to a conventional giant cell tumor of bone . We suggest that this tumor might be categorized in the group of low-grade dedifferentiated chondrosarcomas . Pharmacogenomics of antiplatelet drugs . DB00758 , a platelet Q9H244 inhibitor , is one of the most widely prescribed drugs in cardiovascular medicine because it reduces ischemic and thrombotic complications . It is a prodrug requiring biotransformation into the active metabolite by the hepatic cytochrome 450 system , especially the P33261 enzyme . Candidate gene studies and genome-wide association studies have identified loss-of-function P33261 variants to be associated with a diminished pharmacologic response . Specifically , compared with noncarriers , carriers of at least one copy of a loss-of-function P33261 allele have ∼30 % lower levels of active clopidogrel metabolite and ∼25 % relatively less platelet inhibition with clopidogrel . Moreover , in patients treated with clopidogrel predominantly for percutaneous coronary intervention , carriers of 1 or 2 P33261 loss-of-function alleles are at increased risk for major adverse cardiovascular outcomes , with an ∼1.5-fold increase in the risk of cardiovascular death , myocardial infarction , or stroke as well as an ∼3-fold increase in risk for stent thrombosis . Tripling the dose of clopidogrel in carriers of a P33261 loss-of-function allele can achieve on-treatment platelet reactivity comparable to that seen with the standard 75 mg dose in wild-type individuals , but the impact on clinical outcomes remains unknown . Alternatively , 2 third-generation Q9H244 inhibitors are available : prasugrel and ticagrelor . These drugs are superior to clopidogrel in reducing ischemic outcomes and are unaffected by P33261 loss-of-function alleles . Heroin addiction in African Americans : a hypothesis-driven association study . Heroin addiction is a chronic complex disease with a substantial genetic contribution . This study was designed to identify gene variants associated with heroin addiction in African Americans . The emphasis was on genes involved in reward modulation , behavioral control , cognitive function , signal transduction and stress response . We have performed a case-control association analysis by screening with 1350 variants of 130 genes . The sample consisted of 202 former severe heroin addicts in methadone treatment and 167 healthy controls with no history of drug abuse . Single nucleotide polymorphism ( SNP ) , haplotype and multi-SNP genotype pattern analyses were performed . Seventeen SNPs showed point-wise significant association with heroin addiction ( nominal P < 0.01 ) . These SNPs are from genes encoding several receptors : adrenergic ( ADRA1A ) , arginine vasopressin ( P37288 ) , cholinergic ( P08172 ) , dopamine ( P21728 ) , GABA-A ( P28472 ) , glutamate ( Q12879 ) and serotonin ( P46098 ) as well as alcohol dehydrogenase ( P40394 ) , glutamic acid decarboxylase ( Q99259 and Q05329 ) , the nucleoside transporter ( Q99808 ) and diazepam-binding inhibitor ( DBI ) . The most significant result of the analyses was obtained for the Q12879 haplotype G-A-T ( rs4587976-rs1071502-rs1366076 ) with protective effect ( P(uncorrected) = 9.6E- 05 , P(corrected) = 0.058 ) . This study corroborates several reported associations with alcohol and drug addiction as well as other related disorders and extends the list of variants that may affect the development of heroin addiction . Further studies will be necessary to replicate these associations and to elucidate the roles of these variants in drug addiction vulnerability . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . Partial agonists of the α3β4* neuronal nicotinic acetylcholine receptor reduce ethanol consumption and seeking in rats . DB00898 use disorders ( AUDs ) impact millions of individuals and there remain few effective treatment strategies . Despite evidence that neuronal nicotinic acetylcholine receptors ( nAChRs ) have a role in AUDs , it has not been established which subtypes of the nAChR are involved . Recent human genetic association studies have implicated the gene cluster P32297 - P30532 - P30926 encoding the α3 , α5 , and β4 subunits of the nAChR in susceptibility to develop nicotine and alcohol dependence ; however , their role in ethanol-mediated behaviors is unknown due to the lack of suitable and selective research tools . To determine the role of the α3 , and β4 subunits of the nAChR in ethanol self-administration , we developed and characterized high-affinity partial agonists at α3β4 nAChRs , CP-601932 , and PF-4575180 . Both CP-601932 and PF-4575180 selectively decrease ethanol but not sucrose consumption and operant self-administration following long-term exposure . We show that the functional potencies of CP-601932 and PF-4575180 at α3β4 nAChRs correlate with their unbound rat brain concentrations , suggesting that the effects on ethanol self-administration are mediated via interaction with α3β4 nAChRs . Also varenicline , an approved smoking cessation aid previously shown to decrease ethanol consumption and seeking in rats and mice , reduces ethanol intake at unbound brain concentrations that allow functional interactions with α3β4 nAChRs . Furthermore , the selective α4β2(*) nAChR antagonist , DHβE , did not reduce ethanol intake . Together , these data provide further support for the human genetic association studies , implicating P32297 and P30926 genes in ethanol-mediated behaviors . CP-601932 has been shown to be safe in humans and may represent a potential novel treatment for AUDs . ApoE4 delays dendritic spine formation during neuron development and accelerates loss of mature spines in vitro . The ε4 allele of the gene that encodes apolipoprotein E ( APOE4 ) is the greatest genetic risk factor for Alzheimer 's disease ( AD ) , while APOE2 reduces AD risk , compared to APOE3 . The mechanism(s) underlying the effects of P02649 on AD pathology remains unclear . In vivo , dendritic spine density is lower in APOE4-targeted replacement ( P02649 -TR ) mice compared with APOE2- and APOE3-TR mice . To investigate whether this apoE4-induced decrease in spine density results from alterations in the formation or the loss of dendritic spines , the effects of neuron age and apoE isoform on the total number and subclasses of spines were examined in long-term wild-type neurons co-cultured with glia from APOE2- , APOE3- and APOE4-TR mice . Dendritic spine density and maturation were evaluated by immunocytochemistry via the presence of drebrin ( an actin-binding protein ) with Q05586 ( DB01221 receptor subunit ) and P42262 ( AMPA receptor subunit ) clusters . ApoE isoform effects were analyzed via a method previously established that identifies phases of spine formation ( day-in-vitro , DIV10-18 ) , maintenance ( DIV18-21 ) and loss ( DIV21-26 ) . In the formation phase , apoE4 delayed total spine formation . During the maintenance phase , the density of Q05586 + P42262 spines did not change with apoE2 , while the density of these spines decreased with apoE4 compared to apoE3 , primarily due to the loss of GluA2 in spines . During the loss phase , total spine density was lower in neurons with apoE4 compared to apoE3 . Thus , apoE4 delays total spine formation and may induce early synaptic dysfunction via impaired regulation of GluA2 in spines . Contribution of the P04637 , O15527 , P32297 , and HLA-DQA1 genes to the risk for lung squamous cell carcinoma . INTRODUCTION : Recent genome-wide association studies ( GWASs ) have identified polymorphisms in several genes associated with lung cancer risk . Nevertheless , functional polymorphisms in DNA repair and metabolic genes that had been reported as being associated with risk for lung cancer , particularly for lung squamous cell carcinoma ( SQC ) , were not examined in those studies . Therefore , significance of these functional polymorphisms was evaluated in a population , in which polymorphisms in the GWAS genes showed associations with lung SQC risk . METHODS : Polymorphisms in three DNA repair genes , P04637 , Q00987 , and O15527 , and two metabolic genes , P04798 and P09488 , were examined for associations with lung SQC risk in a hospital-based case-control study consisting of 377 cases and 325 controls , which had been previously subjected to association studies on GWAS genes , P32297 , O14746 , and HLA-DQA1 . RESULTS : Genotypes for two DNA repair genes , P04637 and O15527 , showed significant associations with SQC risk ( p < 0.05 ) , and those for two GWAS genes , P32297 and HLA-DQA1 , showed significant associations with SQC risk ( P < 0.05 ) with odds ratios between 1.65 ( 95 % confidence interval = 1.06-2.57 for O15527 ) and 2.57 ( 95 % confidence interval = 1.03-6.87 for P32297 ) . Marginally significant associations were also observed for Q00987 and P04798 genes . Interactions among these polymorphisms on SQC risk were not observed . CONCLUSIONS : Association of functional polymorphisms in DNA repair and metabolic genes with lung SQC risk was appreciated . This result indicates the necessity of reevaluation for the significance of functional polymorphisms in DNA repair and metabolic genes on lung cancer risk in other populations subjected to GWASs . P10275 and monoamine oxidase polymorphism in wild bonobos . P10275 gene ( AR ) , monoamine oxidase A gene ( P21397 ) and monoamine oxidase B gene ( P27338 ) have been found to have associations with behavioral traits , such as aggressiveness , and disorders in humans . However , the extent to which similar genetic effects might influence the behavior of wild apes is unclear . We examined the loci AR glutamine repeat ( ARQ ) , AR glycine repeat ( ARG ) , P21397 intron 2 dinucleotide repeat ( MAin2 ) and P27338 intron 2 dinucleotide repeat ( MBin2 ) in 32 wild bonobos , Pan paniscus , and compared them with those of chimpanzees , Pan troglodytes , and humans . We found that bonobos were polymorphic on the four loci examined . Both loci MAin2 and MBin2 in bonobos showed a higher diversity than in chimpanzees . Because monoamine oxidase influences aggressiveness , the differences between the polymorphisms of MAin2 and MBin2 in bonobos and chimpanzees may be associated with the differences in aggression between the two species . In order to understand the evolution of these loci and AR , P21397 and P27338 in humans and non-human primates , it would be useful to conduct future studies focusing on the potential association between aggressiveness , and other personality traits , and polymorphisms documented in bonobos . Flip and flop splice variants of AMPA receptor subunits in the spinal cord of amyotrophic lateral sclerosis . Excitotoxicity mediated by AMPA receptors has been suggested to be implicated in the pathogenesis of amyotrophic lateral sclerosis ( P35858 ) . To investigate the relevance of AMPA receptors to motor neuron degeneration in P35858 , we evaluated the expression of mRNAs coding for flip and flop splice variants of AMPA receptor subunits ( P42261 to P48058 ) in the cervical segment of the spinal cord from control individuals and patients with P35858 using in situ hybridization histochemistry . Transcript mRNAs coding for flop variants were significantly decreased in the ventral horn of the spinal cord from patients with P35858 , whereas the mRNAs for flip variants were preserved . These findings suggest that the relative abundance of flip variants vs. flop variants is increased in spinal motor neurons of P35858 patients when compared to that of control individuals . Flip variants promote assemblies of slowly desensitizing AMPA receptors . These results imply that spinal motor neurons of P35858 patients possess more slowly desensitizing AMPA receptors than those of control individuals . This expression change of AMPA receptors in P35858 may account for vulnerability of motor neurons in this disease . Effects of ethanol on the properties of platelets and endothelial cells in model experiments . AIM : To investigate effects of ethanol on activity markers of atherosclerosis in an in vitro endothelial cell model . METHODS : After 24 h incubation with ethanol ( 0.0095 % ) , human umbilical vein endothelial cells were stimulated for 1 h with lipopolysaccharide , and were then incubated in direct contact with activated platelets . Following this incubation , the expression of P29965 and CD62P on platelets , and the expression of intercellular adhesion molecule-1 ( P05362 ) , vascular cell adhesion molecule-1 ( P19320 ) , urokinase plasminogen activator receptor ( Q03405 ) , and membrane-type 1 matrix metalloproteinase ( P50281 ) on endothelial cells were measured by flow cytometry . RESULTS : The increased expression of P19320 and Q03405 on endothelial cells by proinflammatory stimulation with activated platelets was significantly reduced through pre-incubation with ethanol ( P < 0.05 ) . Furthermore , platelets in direct contact with ethanol and with endothelial cells pre-incubated in ethanol showed a significant reduction in their P29965 expression ( P < 0.05 ) . DB00898 had no significant effect on P05362 and P50281 expression on endothelial cells . CONCLUSION : DB00898 directly attenuates platelet activation and has significant endothelial cell-mediated effects on selected markers of atherosclerosis in vitro . These findings underline possible protective effects of ethanol on atherosclerosis .

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interacts_with DB01233?

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P14416 -mediated epidermal growth factor receptor transactivation through a disintegrin and metalloprotease regulates dopaminergic neuron development via extracellular signal-related kinase activation . P14416 ( D2R ) -mediated extracellular signal-regulated kinase ( P29323 ) activation plays an important role in the development of dopaminergic mesencephalic neurons . Here , we demonstrate that D2R induces the shedding of heparin-binding epidermal growth factor ( P01133 ) through the activation of a disintegrin and metalloprotease ( ADAM ) 10 or 17 , leading to P01133 receptor transactivation , downstream P29323 activation , and ultimately an increase in the number of dopaminergic neurons and their neurite length in primary mesencephalic cultures from wild-type mice . These outcomes , however , were not observed in cultures from D2R knock-out mice . Our findings show that D2R-mediated P29323 activation regulates mesencephalic dopaminergic neuron development via P01133 receptor transactivation through O14672 /17 . The relationship between gastric motility and nausea : gastric prokinetic agents as treatments . Nausea is one of a cluster of symptoms described subjectively by patients with delayed gastric emptying . The mechanisms and treatments are unclear ( anti-emetic drugs are not fully effective against nausea ) . Can nausea be relieved by stimulating gastric emptying ? Physostigmine ( together with atropine ) has been shown experimentally to stimulate gastric motility , relieve nausea and restore normal gastric motility . Is this mimicked by gastric prokinetic drugs ? The answer is complicated by mixed pharmacology . DB01233 increases gastric motility by activating myenteric Q13639 receptors but also directly inhibits vomiting via D2 and 5- Q9H205 receptor antagonism ; relationships between increased gastric motility and relief from nausea are therefore unclear . Similarly , the D2 receptor antagonist domperidone has direct anti-emetic activity . Nevertheless , more selective Q13639 and motilin receptor agonists ( erythromycin , directly stimulating gastric motility ) inhibit vomiting in animals ; low doses of erythromycin can also relieve symptoms in patients with gastroparesis . Ghrelin stimulates gastric motility and appetite mostly via vagus-dependent pathways , and inhibits vomiting in animals . To date , ghrelin receptor activation has failed to consistently improve gastric emptying or symptoms in patients with gastroparesis . We conclude that nausea can be relieved by gastric prokinetic drugs , but more clinical studies are needed using drugs with selective activity . Other mechanisms ( e.g. ghrelin , vagal and central pathways , influencing a mechanistic continuum between appetite and nausea ) also require exploration . These and other issues will be further explored in a forthcoming special issue of the European Journal of Pharmacology , which focusses on mechanisms of nausea and vomiting . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Effects of metoclopramide and tropisetron on aldosterone secretion possibly due to agonism and antagonism at the Q13639 receptor . OBJECTIVE : Part of the prokinetic activity of metoclopramide can possibly be ascribed to agonist activity at Q13639 receptors . The 5- Q9H205 antagonist tropisetron is thought to act as an antagonist at Q13639 receptors . In the present study aldosterone secretion in response to the administration of these two drugs was explored to examine the role of the Q13639 receptor in aldosterone secretion . METHODS : Following a single-blind , random design , ten normal male volunteers received one of the following regimens on three occasions , with at least 2-week intervals : metoclopramide 10 mg i.v. ; tropisetron 5 mg by slow i.v.i. , or ; tropisetron by slow i.v.i. , followed by 10 mg metoclopramide i.v. RESULTS : In response to metoclopramide alone the mean plasma aldosterone level rose significantly to 149 % of basal level and remained significantly elevated for the next 20 min . With tropisetron alone , there was a significant 37.8 % drop at 60 min and the aldosterone levels remained low for the duration of the experiment . DB01233 reversed the decline mediated by tropisetron significantly at 30 and 90 min . DB04630 levels after the latter regimen also did not differ significantly from baseline at any time period . CONCLUSION : These results would suggest the existence of a tonic stimulatory influence of 5-HT via Q13639 receptors on aldosterone secretion , which could be augmented by metoclopramide and blocked by tropisetron . However , the effect of tropisetron per se should be interpreted with caution given the lack of a saline group . Effect of metoclopramide , ondansetron and granisetron on aldosterone secretion in man . The plasma aldosterone response following the administration of drugs with antagonist and agonist activity at Serotonin 3 and 4 ( 5- Q9H205 & 4 ) receptors has been examined in 9 healthy male volunteers receiving the following four treatments i.v. in a randomised , cross-over sequence : ondansetron 8 mg , granisetron 3 mg , metoclopramide 20 mg , and saline 20 ml . DB01233 significantly increased the mean plasma aldosterone level to 196 % of basal level at 5 min . It rose to 234 % at 15 min and remained at more than 185 % of basal level for the duration of the experiment . The response to ondansetron and granisetron did not differ significantly from placebo . If dopamine antagonism is discounted , the results suggest that metoclopramide-induced aldosterone secretion results from its agonist activity at Q13639 receptors , although slow neuronal depolarization via an unidentified receptor remains a possibility . Antagonism at the 5- Q9H205 receptor plays no role , as the selective antagonist , granisetron , did not elicit a significant response . It seems unlikely that the Q13639 receptor is the second , low affinity binding site of ondansetron , unless it had no agonist activity at this receptor . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Synthesis and characterization of the first fluorescent antagonists for human Q13639 receptors . Fluorescent antagonists for human 5-HT(4) receptors were synthesized based on ML10302 1 , a potent 5-HT(4) receptor agonist and on piperazine analogue 2 . These molecules were derived with three fluorescent moieties , dansyl , naphthalimide , and NBD ( 7-nitrobenz-2-oxa-1,3-diazol-4-yl ) , through alkyl chains . The synthesized molecules were evaluated in binding assays on the recently cloned human 5-HT(4(e)) receptor isoform stably expressed in P13671 glial cells with [(3)H]GR113808 as the radioligand . The affinity values depended upon the basal structure together with the alkyl chain length . The derivatives based on ML10302 were more potent ligands than the derivatives based on piperazine analogue . For ML10302-based ligands , dansyl and NBD derivatives attached through a chain length of one carbon atom 17a and 32 , respectively , led to affinities close to the affinity of ML10302 . The most potent compounds 17a , 28 , and 32 produced an inhibition of the 5-HT stimulated cyclic AMP synthesis in the same cellular system with nanomolar K(b) values . Fluorescent properties of 17a , 28 , and 32 were more particularly studied . Interactions of the fluorescent ligand 28 with the h5-HT(4(e)) receptor were indicated using h5-HT(4(e)) receptor transfected P13671 glial cell membranes and entire cells . Ligand 28 was also used in fluorescence microscopy experiments in order to label h5-HT(4(e)) receptor transfected P13671 glial cells , and subcellular localization of these receptors was more precisely determined using confocal microscopy . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . Effects of cholinoceptor and 5-hydroxytryptamine3 receptor antagonism on erythromycin-induced canine intestinal motility disruption and emesis . 1. Erythromycin administration is associated with gastrointestinal problems , disturbed gastrointestinal motility and emesis . This study in the dog investigates the underlying mechanisms . 2 . Intestinal myoelectrical activity and the occurrence and latency of emesis were recorded in eight conscious dogs . All drugs were administered intravenously . 3 . Erythromycin ( 7 mg kg-1 ) increased contractions of the proximal small intestine , and caused emesis in all fasted dogs and in 5 dogs after food . Atropine ( 50 mg kg-1 min-1 ) and hexamethonium ( 10 mg kg-1 h-1 ) partially inhibited the GI motility effects but did not significantly reduce emesis . 4 . DB01233 at a high dose ( 2 mg kg-1 h-1 ) reduced the incidence of emesis in the presence of increased intestinal motility , but a low dose ( 150 micrograms kg-1 h-1 ) was ineffective . 5 . A 5-hydroxytryptamine3 ( 5- Q9H205 ) receptor antagonist , MDL 72222 ( 1 mg kg-1 ) , reduced emesis when given alone and combined with metoclopramide ( low dose ) . The Q13639 receptor agonist BRL24924 ( DB04917 , 1 mg kg-1 ) had no effect on emesis either alone in combination with metoclopramide . 6 . In conclusion , erythromycin-induced GI motility disturbances and emesis are not causally related . Whereas the increase in intestinal smooth muscle activity is possibly cholinergically mediated , emesis occurs at least in part via a 5-hydroxytryptaminergic mechanism , but does not involve the dopamine system . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . DB01233 does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 ( MCP ) , the most frequently used prokinetic drug in neonates , enhances gastric muscle contractility through inhibition of dopamine receptors . Although its therapeutic benefit is established in adults , limited data are available to support its clinical use in infants . Hypothesizing that developmentally dependent differences are present , we comparatively evaluated the effect of MCP on fundus muscle contractility in newborn , juvenile , and adult rats . The muscle strips were either contracted with electrical field stimulation ( O43281 ) to induce cholinergic nerve-mediated acetylcholine release or carbachol , a cholinergic agonist acting directly on the muscarinic receptor . Although in adult rats MCP increased O43281 -induced contraction by 294 ± 122 % of control ( P < 0.01 ) , no significant effect was observed in newborn fundic muscle . MCP had no effect on the magnitude of the carbachol-induced and/or bethanechol-induced gastric muscle contraction at any age . In response to dopamine , an 80.7 ± 5.3 % relaxation of adult fundic muscle was observed , compared with only a 8.4 ± 8.7 % response in newborn tissue ( P < 0.01 ) . P14416 expression was scant in neonates and significantly increased in adult gastric tissue ( P < 0.01 ) . In conclusion , the lack of MCP effect on the newborn fundic muscle contraction potential relates to developmental differences in dopamine D2 receptor expression . To the extent that these novel data can be extrapolated to neonates , the therapeutic value of MCP as a prokinetic agent early in life requires further evaluation . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . P14416 stimulation of Na+/H+ exchange assessed by quantification of extracellular acidification . A microphysiometer was used to quantify the rate of extracellular acidification by P13671 glioma cells and L fibroblasts expressing recombinant dopamine D2 receptors . The dopamine D2 receptor agonist , quinpirole , accelerated the rate of acidification of the medium by P13671 cells expressing either the short or long form of D2 receptors , D2(415) and D2(444) , but not by wild-type cells that were not transfected with a D2 receptor cDNA . The rate of acidification increased with increasing concentrations of quinpirole up to 100 nM . Inhibition of the response by the dopamine D2 antagonist , spiperone , provided additional evidence that the enhanced extracellular acidification resulted from stimulation of D2 receptors . To test the hypothesis that D2 receptor-stimulated extracellular acidification was due to transport of protons by a Na+/H+ antiporter and reflected intracellular alkalinization , the effect of two inhibitors of Na+/H+ exchange , amiloride and methyl-isobutyl-amiloride , was determined . Both compounds inhibited quinpirole-induced extracellular acidification at concentrations that did not alter D2 receptor-mediated inhibition of adenylylcyclase or radioligand binding to D2 receptors . In addition , quinpirole-induced extracellular acidification was greatly inhibited by removal of sodium from the extracellular medium , confirming the participation of Na+/H+ exchange in the extrusion of acid . Quinpirole ( 100 nM ) also increased the rate of extracellular acidification by L cells expressing D2(415) , LZR1 cells . Treatment with pertussis toxin ( 100 ng/ml for 18 h ) had no effect on the quinpirole-induced acid extrusion by C6D2(415) and LZR1 cells , although the same pertussis toxin treatment regimen completely prevented inhibition of adenylylcyclase . We conclude that recombinant D2 receptors accelerate Na+/H+ exchange in P13671 cells and L fibroblasts by a pathway that does not involve inhibition of adenylylcyclase or pertussis toxin-sensitive G proteins . DB01233 stimulates catecholamine- and granin-derived peptide secretion from pheochromocytoma cells through activation of serotonin type 4 ( Q13639 ) receptors . The gastroprokinetic agent metoclopramide is known to stimulate catecholamine secretion from pheochromocytomas . The aim of the study was to investigate the mechanism of action of metoclopramide and expression of serotonin type 4 ( 5-HT(4) ) receptors in pheochromocytoma tissues . Tissue explants , obtained from 18 pheochromocytomas including the tumor removed from a 46-year-old female patient who experienced life-threatening hypertension crisis after metoclopramide administration and 17 additional pheochromocytomas ( 9 benign and 8 malignant ) were studied . Cultured pheochromocytoma cells derived from the patient who previously received metoclopramide were incubated with metoclopramide and various 5-HT(4) receptor ligands . In addition , total mRNAs were extracted from all the 18 tumors . Catecholamine- and granin-derived peptide concentrations were measured in pheochromocytoma cell incubation medium by HPLC and radioimmunological assays . In addition , expression of 5-HT(4) receptor mRNAs in the 18 pheochromocytomas was investigated by the use of reverse transcriptase-PCR . RESULTS : DB01233 and the 5-HT(4) receptor agonist cisapride were found to activate catecholamine- and granin-derived peptide secretions by cultured tumor cells . DB01233 - and cisapride-evoked catecholamine- and granin-derived peptide productions were inhibited by the 5-HT(4) receptor antagonist GR 113808 . 5-HT(4) receptor mRNAs were detected in the patient 's tumor and the series of 17 additional pheochromocytomas . This study shows that pheochromocytomas express functional 5-HT(4) receptors that are responsible for the stimulatory action of metoclopramide on catecholamine- and granin-derived peptide secretion . All 5-HT(4) receptor agonists must therefore be contraindicated in patients with proven or suspected pheochromocytoma . Endogenous serotonin excites striatal cholinergic interneurons via the activation of 5-HT 2C , P50406 , and P34969 serotonin receptors : implications for extrapyramidal side effects of serotonin reuptake inhibitors . The striatum is richly innervated by serotonergic afferents from the raphe nucleus . We explored the effects of this input on striatal cholinergic interneurons from rat brain slices , by means of both conventional intracellular and whole-cell patch-clamp recordings . Bath-applied serotonin ( 5-HT , 3-300 microM ) , induced a dose-dependent membrane depolarization and increased the rate of spiking . This effect was mimicked by the 5-HT reuptake blockers citalopram and fluvoxamine . In voltage-clamped neurons , 5-HT induced an inward current , whose reversal potential was close to the K(+) equilibrium potential . Accordingly , the involvement of K(+) channels was confirmed either by increasing extracellular K(+) concentration and by blockade of K(+) channels with barium . Single-cell reverse transcriptase-polymerase chain reaction ( RT-PCR ) profiling demonstrated the presence of P28335 , P50406 , and P34969 receptor mRNAs in identified cholinergic interneurons . The depolarization/inward current induced by 5-HT was partially mimicked by the 5-HT2 receptor agonist 2,5-dimethoxy-4-iodoamphetamine and antagonized by both ketanserin and the selective P28335 antagonist RS102221 , whereas the selective 5- Q9H205 and Q13639 receptor antagonists tropisetron and RS23597-190 had no effect . The depolarizing response to 5-HT was also reduced by the selective P50406 and P34969 receptor antagonists SB258585 and SB269970 , respectively , and mimicked by the P34969 agonist , 5-CT . Accordingly , activation of either P50406 or P34969 receptor induced an inward current . The 5-HT response was attenuated by U73122 , blocker of phospholipase C , and by SQ22,536 , an inhibitor of adenylyl cyclase . These results suggest that 5-HT released by serotonergic fibers originating in the raphe nuclei has a potent excitatory effect on striatal cholinergic interneurons . Analysis of neurogenic contractions induced by ML-1035 and other benzamides in the guinea-pig non-stimulated isolated ileum . 4-Amino-5-chloro-substituted benzamides have been shown to increase gastric motility in-vivo and enhance field-stimulated and peristaltic contractions in-vitro . The present experiments examined the contractile response to a series of benzamides in the guinea-pig non-stimulated ileum . Four benzamides elicited contractions in the isolated ileum which were expressed as a percentage of the contraction induced by 1 microM acetylcholine ( % acetylcholine response = 12 +/- 2 , 19 +/- 3 , 26 +/- 2 , 51 +/- 3 , n = 13 , 8 , 17 , and 21 , with EC50 values of 0.85 , 1.8 , 5.7 , and 14.2 microM for cisapride , zacopride , metoclopramide , and ML-1035 ( 4-amino-5-chloro-2-((2-methylsulphinyl)-ethoxy)-N- ( 2-(diethylamino)-ethyl ) -benzamide hydrochloride ) , respectively ) . ML-1035 contractions were completely blocked by atropine and tetrodotoxin , while ganglionic blockade with hexamethonium was ineffective . DB01233 has been reported to sensitize postjunctional muscarinic receptors , however , ML-1035 did not enhance acetylcholine-induced contractions . Tropisetron ( ICS 205-930 , 1 microM ) , caused a parallel rightward shift in the concentration-response curve for both ML-1035 and zacopride ( EC50 = 14.2 +/- 1.3 and 1.8 +/- 0.8 microM in the absence , and 26 +/- 2.7 and 6.9 +/- 2.3 microM in the presence of tropisetron for ML-1035 and zacopride , respectively ) with apparent pKB values of 5.9 and 6.0 for the respective compounds . 5-Hydroxytryptaminergic receptor desensitization by 2-methyl-5-hydroxytryptamine ( 5- Q9H205 ) and 5-methoxytryptamine ( Q13639 ) , attenuated the response to ML-1035. ( ABSTRACT TRUNCATED AT 250 WORDS )

MH_train_1014

MH_train_1014

MH_train_1014

interacts_with DB00030?

multiple_choice

Molecular markers for novel therapeutic strategies in pancreatic endocrine tumors . OBJECTIVES : Pancreatic endocrine tumors ( PETs ) share numerous features with gastrointestinal neuroendocrine ( carcinoid ) tumors . Targets of novel therapeutic strategies previously assessed in carcinoid tumors were analyzed in PETs ( 44 cases ) . METHODS : Activating mutations in P00533 , P10721 , and P16234 and nonresponse mutations in P01116 were evaluated . Copy number of P00533 and HER-2/neu was quantified by fluorescence in situ hybridization . Expression of P00533 , P16234 , P17948 , P36897 , Hsp90 , SSTR2A , P35346 , P08069 , P42345 , and P16455 was measured immunohistochemically . RESULTS : Elevated P00533 copy number was found in 38 % of cases but no P01116 nonresponse mutations . P17948 , P36897 , P16234 , P35346 , SSTR2A , and P08069 exhibited the highest levels of expression in the largest percentages of PETs.Anticancer drugs BMS-754807 ( selective for P08069 /IR ) , 17-(allylamino)-17-demethoxygeldanamycin ( 17- P29372 , targeting Hsp90 ) , and axitinib ( directed toward P17948 -3/ P16234 -B/ P10721 ) induced growth inhibition of human QGP-1 PET cells with IC50 values ( nM ) of 273 , 723 , and 743 , respectively . At growth-inhibiting concentrations , BMS-754807 inhibited P08069 phosphorylation ; 17- P29372 induced loss of P00533 , P08069 , and P35968 ; and axitinib increased P38936 ( P38936 ) expression without inhibiting P35968 phosphorylation . CONCLUSIONS : Results encourage further research into multidrug strategies incorporating inhibitors targeting P08069 or Hsp90 and into studies of axitinib combined with conventional chemotherapeutics toxic to tumor cells in persistent growth arrest . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Readjusting the localization of long QT syndrome gene on chromosome 11p15 . Long QT syndrome ( LQT ) is an autosomal dominant cardiac disease characterized by ventricular arrhythmia . A first locus for LQT has been identified on chromosome 11p15.5 ( LQT1 ) , closely linked to P01112 . To refine the location of LQT1 , microsatellites were genotyped in 8 French families and the following order was determined : tel- P01112 - P21917 -D11S922-D11S4046- P01344 - P01308 -TH-D11S1318-D11S1323-D11S1338-D11S90 9-D11S1346-cen . By haplotype analysis , 12 crossing-over events were identified in affected and unaffected subjects , delineating the LQT1 candidate region to 7 cM . This new delineated localization between D11S1318 and D11S1323 is in a more centromeric region than previously thought and is 5 cM proximal to P01112 . Immunohistochemical detection of alpha1E voltage-gated Ca(2+) channel isoforms in cerebellum , P01308 -1 cells , and neuroendocrine cells of the digestive system . Polyclonal antibodies were raised against a common and a specific epitope present only in longer alpha1E isoforms of voltage-gated Ca(2+) channels , yielding an " anti-E-com " and an " anti-E-spec " serum , respectively . The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected P29320 -293 cells or membrane proteins derived from them . Cells from the insulinoma cell line P01308 -1 , tissue sections from cerebellum , and representative regions of gastrointestinal tract were stained immunocytochemically . P01308 -1 cells expressed an alpha1E splice variant with a longer carboxy terminus , the so-called alpha1Ee isoform . Similarily , in rat cerebellum , which was used as a reference system , the anti-E-spec serum stained somata and dendrites of Purkinje cells . Only faint staining was seen throughout the cerebellar granule cell layer . After prolonged incubation times , neurons of the molecular layer were stained by anti-E-com , suggesting that a shorter alpha1E isoform is expressed at a lower protein density . In human gastrointestinal tract , endocrine cells of the antral mucosa ( stomach ) , small and large intestine , and islets of Langerhans were stained by the anti-E-spec serum . In addition , staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae . We conclude that the longer alpha1Ee isoform is expressed in neuroendocrine cells of the digestive system and that , in pancreas , alpha1Ee expression is restricted to the neuroendocrine part , the islets of Langerhans. alpha1E therefore appears to be a common voltage-gated Ca(2+) channel linked to neuroendocrine and related systems of the body . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . P01308 reverses growth hormone-induced homologous desensitization . Growth hormone ( GH ) is secreted in a pulsatile pattern to promote body growth and metabolism . GH exerts its function by activating several signaling pathways , including O60674 / P35610 and MEK/ P29323 . P27361 /2 activation by GH plays important roles in gene expression , cell proliferation , and growth . We previously reported that in rat H4IIE hepatoma cells after an initial GH exposure , a second GH exposure induces P42229 phosphorylation but not P27361 /2 phosphorylation ( Ji , S. , Frank , S. J. , and Messina , J. L. ( 2002 ) J. Biol. Chem. 277 , 28384-28393 ) . In this study the mechanisms underlying GH-induced homologous desensitization were investigated . A second GH exposure activated the signaling intermediates upstream of MEK/ P29323 , including O60674 , Ras , and P04049 . This correlated with recovery of P10912 levels , but was insufficient for GH-induced phosphorylation of Q02750 /2 and P27361 /2 . P01308 restored the ability of a second GH exposure to induce phosphorylation of Q02750 /2 and P27361 /2 without altering P10912 levels or GH-induced phosphorylation/activation of O60674 and P04049 . GH and insulin synergized in promoting cell proliferation . Further investigation suggested that insulin increased the amount of MEK bound to Q8IVT5 ( kinase suppressor of Ras ) and restored GH-induced tyrosine phosphorylation of Q8IVT5 . Previous GH exposure also induced desensitization of P42224 and P40763 phosphorylation , but this desensitization was not reversed by insulin . Thus , insulin-regulated resensitization of GH signaling may be necessary to reset the complete response to GH after a normal , physiologic pulse of GH . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . Q99717 regulates Akt2 expression and insulin-induced glucose uptake in Q9BTT4 myotubes . P01308 -induced glucose uptake by skeletal muscle results from Akt2 activation and is severely impaired during insulin resistance . Recently , we and others have demonstrated that Q9UK05 improves glucose homeostasis in diabetic and non-diabetic rodents . However , the mechanism by which Q9UK05 modulates insulin action remains unknown . Here we demonstrate that Q99717 , a transcription factor activated by Q9UK05 , and Akt2 , are upregulated in differentiated Q9BTT4 myotubes . Q99717 , rather than Q15797 /8 , is downregulated " in vivo " and " in vitro " by dexamethasone . Q99717 knockdown decreased Akt2 expression and serine phosphorylation and insulin-induced glucose uptake , and increased the expression of the lipid phosphatase Ship2 . Additionally , binding of Q99717 to Akt2 gene is decreased in dexamethasone-treated rats and increased in Q9BTT4 myotubes compared to myoblasts . The present study indicates that Q99717 regulates glucose uptake in skeletal muscle by controlling Akt2 expression and phosphorylation . These finding reveals Q99717 as a potential target for the therapeutic of type 2 diabetes . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . Survey of differentially methylated promoters in prostate cancer cell lines . DNA methylation and copy number in the genomes of three immortalized prostate epithelial and five cancer cell lines ( LNCaP , PC3 , PC3M , PC3M-Pro4 , and PC3M-LN4 ) were compared using a microarray-based technique . Genomic DNA is cut with a methylation-sensitive enzyme HpaII , followed by linker ligation , polymerase chain reaction ( PCR ) amplification , labeling , and hybridization to an array of promoter sequences . Only those parts of the genomic DNA that have unmethylated restriction sites within a few hundred base pairs generate PCR products detectable on an array . Of 2732 promoter sequences on a test array , 504 ( 18.5 % ) showed differential hybridization between immortalized prostate epithelial and cancer cell lines . Among candidate hypermethylated genes in cancer-derived lines , there were eight ( P16070 , P38936 , P03372 , P00749 , P10826 , SFN , P25445 , and Q01534 ) previously observed in prostate cancer and 13 previously known methylation targets in other cancers ( O95661 , bcl-2 , P38398 , P42773 , P24522 , Q13126 , P06401 , O43511 , P09486 , P43405 , Q9UDY2 , P09936 , and Q06250 ) . The majority of genes that appear to be both differentially methylated and differentially regulated between prostate epithelial and cancer cell lines are novel methylation targets , including Q9NQU5 , Q92878 , O43711 , Q96B01 , Q13163 , P06213 , P35555 , and GG2-1 , representing a rich new source of candidate genes used to study the role of DNA methylation in prostate tumors . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Concise prediction models of anticancer efficacy of 8 drugs using expression data from 12 selected genes . We developed concise , accurate prediction models of the in vitro activity for 8 anticancer drugs ( DB00544 , DB00515 , DB00305 , DOX , CPT-11 , SN-38 , TXL and TXT ) , along with individual clinical responses to DB00544 using expression data of 12 genes . We first performed cDNA microarray analysis and MTT assay of 19 human cancer cell lines to sort out genes which were correlative in expression levels with cytotoxicities of the 8 drugs ; we selected 13 genes with proven functional significance to drug sensitivity from a huge number of potent prediction marker genes . The correlation significance of each was confirmed using expression data quantified by real-time RT-PCR , and finally 12 genes ( P08183 , Q9UNQ0 , P10632 , P08684 , Q12882 , P09211 , P16455 , P15559 , P16435 , P11388 , P07437 and P04818 ) were selected as more reliable predictors of drug response . Using multiple regression analysis , we fixed 8 prediction formulae which embraced the variable expressions of the 12 genes and arranged them in order , to predict the efficacy of the drugs by referring to the value of Akaike 's information criterion for each sample . These formulae appeared to accurately predict the in vitro efficacy of the drugs . For the first clinical application model , we fixed prediction formulae for individual clinical response to DB00544 in the same way using 41 clinical samples obtained from 30 gastric cancer patients and found to be of predictive value in terms of survival , time to treatment failure and tumor growth . None of the 12 selected genes alone could predict such clinical responses .

MH_train_1015

MH_train_1015

MH_train_1015

interacts_with DB01017?

multiple_choice

Matrix metalloproteinase ( MMP ) -9 , but not P08253 , is involved in the development and progression of C protein-induced myocarditis and subsequent dilated cardiomyopathy . Repeated or continuous inflammation of the heart is one of the initiation factors for dilated cardiomyopathy ( DCM ) . In previous studies , we established a DCM animal model by immunizing rats with cardiac C protein . In the present study , we analyze the role of matrix metalloproteinases ( MMPs ) in experimental autoimmune carditis ( EAC ) and subsequent DCM to elucidate the pathomechanisms of this disease . In this model , inflammation begins approximately 9 days after immunization . At that time , MMP activities were detected by in situ zymography . Real-time PCR analysis revealed continuous up-regulation of P08253 mRNA from 2 wk and thereafter . P14780 mRNA , however , had only a transient increase at 2 wk . Double staining with in situ zymography and cell markers demonstrated that gelatinase ( P08253 and P14780 ) -expressing cells are infiltrating macrophages during the early stage and cardiomyocytes at later stages . DB01017 , which inhibits P14780 activities more strongly than P08253 , significantly suppressed EAC , but an P08253 -specific inhibitor , TISAM , did not affect the course of the disease . Furthermore , immunohistochemical examination revealed that minocycline treatment suppressed T cell and macrophage infiltration strongly , whereas TISAM did not . These findings indicate that P14780 , but not P08253 , is involved in the pathogenesis of the acute phase of EAC , and further suggest that P14780 inhibitors , minocycline and its derivatives , may be useful therapies for EAC and DCM . Chemotactic and cytotoxic effects of minocycline on human retinal pigment epithelial cells . PURPOSE : To reveal the effects of minocycline , an anti-inflammatory and neuroprotective agent , on the viability and physiological properties of retinal pigment epithelial ( Q96AT9 ) cells and to compare the effects with those of triamcinolone acetonide . METHODS : The proliferation of human Q96AT9 cells in vitro was investigated with a bromodeoxyuridine immunoassay ; chemotaxis was examined with a Boyden chamber assay . Cell viability was determined by trypan blue exclusion . The gene expression of growth factors and P14780 was determined with real-time RT-PCR , and the secretion of P15692 was examined with ELISA . The phosphorylation of p38 MAPK and P27361 /2 proteins was determined with Western blot analysis . RESULTS : DB01017 at low concentrations ( 50 nM-20 microM ) stimulated chemotaxis and decreased the proliferation of Q96AT9 cells . DB01017 at high concentrations ( above 5 microM ) decreased the viability of Q96AT9 cells through the induction of cell necrosis . The chemotactic effect of minocycline was mediated by the stimulation of autocrine PDGF signaling and the activation of p38 MAPK . DB01017 promoted the expression of PDGF-B , P14210 , P15692 , and P14780 and increased the amounts of phosphorylated p38 and P27361 /2 proteins in Q96AT9 cells . DB00620 reduced PDGF-evoked chemotaxis and P15692 expression and secretion and had no significant effects on cell viability and proliferation . DB00620 did not reverse the effects of minocycline on cell proliferation , chemotaxis , or viability or the expression of P15692 . CONCLUSIONS : Low-dose minocycline induces the activation of Q96AT9 cells , as indicated by the activation of p38 and P27361 /2 and by enhanced chemotaxis mediated by autocrine PDGF signaling . High-dose minocycline induces Q96AT9 cell degeneration . DB01017 exerts multiple inhibitory effects on vascular endothelial growth factor-induced smooth muscle cell migration : the role of P27361 /2 , PI3K , and matrix metalloproteinases . Widely used tetracycline antibiotics affect many cellular functions relevant to human vascular disease including cell proliferation , migration , and matrix remodeling . We examined whether minocycline inhibited human aortic smooth muscle cell ( HASMC ) migration induced by vascular endothelial growth factor ( P15692 ) . After the establishment of an optimal dose , minocycline treated HASMC were exposed to P15692 . HASMC migration , matrix metalloproteinase ( MMP ) -2 and P14780 activities , mitogen-activated protein kinase ( MAPK ) , and phosphatidylinositol 3-kinase ( PI3K ) phosphorylation were determined by smooth muscle cell ( SMC ) invasion assay , real-time polymerase chain reaction , zymograms , and Western blot analysis , respectively . We demonstrated that P15692 and platelet-derived growth factor ( PDGF ) -induced SMC migration in a dose-dependent manner . P14780 , but not P08253 , mRNA was increased during P15692 stimulation . P14780 activity was increased from 1.5- to 2.5-fold in a dose-dependent manner ( P < 0.05 ) . Both P27361 /2 and PI3K/AKt pathways were activated during P15692 -induced HASMCs migration . We then demonstrated that minocycline can inhibit P15692 -induced HASMC migration ( P < 0.05 ) . The effects may be through the inhibition of P14780 mRNA transcription , protein activities and downregulation of P27361 /2 and PI3K/Akt pathway phosphorylation . Our results indicated that minocycline exerts multiple effects on P15692 -induced SMC migration , including inhibition of P14780 mRNA transcription and protein activities and downregulating P27361 /2 and PI3K signal pathways , suggesting minocycline may be a potentially therapeutic approach to inhibit disease process induced angiogenesis . DB01017 with aspirin : an approach to attenuate diabetic nephropathy in rats . Degradation of extracellular matrix ( Q13201 ) by enhanced production of matrix metalloproteinase-2 ( P08253 ) and matrix metalloproteinase-9 ( P14780 ) in diabetes leads to nephropathy . Cyclooxygenases ( P36551 ) further increase levels of these MMPs . The objective of present study was to inhibit P08253 and P14780 by combination of minocycline and aspirin to treat diabetic nephropathy . Diabetes was induced in male Wistar rats by streptozotocin ( Q11206 , 55 mg/kg i.p. ) . Four weeks after diabetes induction , the rats were treated with minocycline ( 50 mg/kg , p.o. ) , aspirin ( 50 mg/kg , p.o. ) , or minocycline ( 50 mg/kg , p.o. ) plus aspirin ( 50 mg/kg , p.o. ) for a period of 4 weeks . At the end of eighth week fluid input , urine output , and renal function tests were carried out for diagnosis of diabetic nephropathy . Renal hypertrophy was measured and histopathology was done to evaluate renal damage . Diabetes produced significant loss of body weight , polyuria , polydipsia , hyperglycemia , and increase in blood pressure . Serum creatinine , urea , and blood urea nitrogen levels were found to be increased significantly in the Q11206 group diabetic rats . Treatment with combination of minocycline and aspirin significantly prevented the rise in creatinine , urea , and blood urea nitrogen levels and increased creatinine clearance . Image analysis of kidneys revealed that collagen level was significantly decreased in combined treated group when compared with control . Results of present study suggest that P08253 and P14780 inhibition in presence of P36551 inhibitor prevents the development of experimental diabetic nephropathy in rats and can be a potential approach for the treatment . DB01197 attenuates matrix metalloproteinase-2 and -9 in monocrotaline-induced right ventricular hypertrophy in rats . Little is known about the influence of angiotensin converting enzyme ( P12821 ) inhibitors on matrix metalloproteinase ( MMP ) in right ventricular remodeling . We investigated the effect of captopril , an P12821 inhibitor , on P08253 and P14780 in monocrotaline-induced right ventricular hypertrophy . Six-week-old male Wistar rats were injected intraperitoneally with monocrotaline ( 60 mg/kg ) or saline . The rats were administrated captopril ( 30 mg/kg per day ) or a vehicle orally for 24 days from the day of monocrotaline injection . At day 25 , echocardiography was performed and hearts were excised . Expressions and activities of P08253 and P14780 were measured by Western blotting and by gelatin zymography , respectively . In monocrotaline-injected rats , right ventricular weight/tail length ratio increased significantly . Histological analysis revealed cardiomyocyte hypertrophy and fibrosis in right ventricular sections . Echocardiography showed right ventricular dysfunction compared with saline-injected rats . The right ventricular hypertrophy , fibrosis , and dysfunction were inhibited by captopril . However , captopril did not attenuate an increase in pulmonary artery pressure . P08253 and P14780 expressions and activities in right ventricles increased significantly in monocrotaline-injected rats and captopril inhibited them . These findings indicate that captopril attenuates the development of monocrotaline-induced right ventricular hypertrophy in association with inhibition of P08253 and P14780 in rats . Influence of polymorphisms and P01375 and IL1β serum concentration on the infliximab response in Crohn 's disease and ulcerative colitis . AIM : Inflammatory bowel diseases ( Q9UKU7 ) , such as Crohn 's disease ( CD ) and ulcerative colitis ( UC ) , are partially attributable to an increased secretion of proinflamatory cytokines , such as tumour necrosis factor ( P01375 ) and interleukin-1β ( IL1β ) , which play essential roles in the disease pathogenesis and are target molecules for specific therapy . Given the inter-individual variability in the response to the anti- P01375 monoclonal antibody infliximab , the aim of our study was to explore the predictive value of P01375 and/or IL1β as surrogate markers of infliximab response . METHODS : Serial serum concentrations of P01375 and IL1β and P01375 promoter region and P01584 polymorphisms were determined in 47 patients ( 29 CD and 18 UC ) receiving infliximab and correlated with treatment response . RESULTS : Baseline serum concentrations of P01375 and IL1β were higher in UC patients than in CD patients ( p = 0.0097 and 0.0024 , respectively ) . CD patients showing < 0.64 pg/ml IL1β at baseline were more frequently responders than non-responders ( p = 0.036 ) , and the C allele of the P01584 polymorphism was associated with higher IL1β serum concentrations ( p = 0.026 ) and with poorer clinical remission after 14 weeks of infliximab treatment . No significant association was found between serum P01375 concentration or P01375 polymorphism and patient response to infliximab . CONCLUSION : This is the first study evaluating the pharmacogenetic role of the rs1143634 polymorphism of P01584 and P01375 polymorphisms in infliximab-treated Q9UKU7 patients . We found an association between the rs1143634 C allele and higher serum IL1β concentrations and a lower response to infliximab treatment in CD patients that warrants the interest of future studies in larger and independent series . Canine pre-iridal fibrovascular membranes : morphologic and immunohistochemical investigations . OBJECTIVE : Pathologic intraocular neovascularization is a key component of many canine ophthalmic diseases such as uveitis , retinal detachment , intraocular neoplasms , and corneal perforation . The purpose of this study was to evaluate the structure of pre-iridal fibrovascular membranes ( PIFMs ) associated with several different disease processes and to identify specific factors associated with their development in the canine eye . PROCEDURE : This study examined 36 enucleated canine eyes with the diagnosis of PIFM and one of the following : lens-induced uveitis , retinal detachment , iridociliary adenoma , corneal perforation , severe hyphema , or vitreal gliovascular membranes ( canine ocular gliovascular syndrome , COGS ) . Three histologic stains and six immunohistochemical stains were performed in all 36 PIFM eyes and four histologically normal eyes , including : hematoxylin and eosin , alcian blue periodic acid schiff ( DB00233 ) , Masson 's trichrome , platelet endothelial cell adhesion molecule-1 ( CD31 ) , smooth muscle actin , vimentin , laminin , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) . RESULTS : Pre-iridal fibrovascular membrane extracellular matrix staining was consistent with collagen and mucins in all cases and positive for laminin in most cases . All PIFMs contained CD31-positive vessels and predominantly lymphoplasmacytic inflammation . Both PIFM vessels and spindle cells were positive for laminin , vimentin , smooth muscle actin , P15692 , and P35354 . Secondary intraocular pathology and immunohistochemical staining of other intraocular structures are also reported . CONCLUSIONS : Pre-iridal fibrovascular membrane morphology and immunohistochemical characteristics were similar across six canine disease processes , suggesting analogous pathophysiologic mechanisms . P35354 and P15692 were identified using immunohistochemistry and may play a role in PIFM development . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . DB01017 protects Schwann cells from ischemia-like injury and promotes axonal outgrowth in bioartificial nerve grafts lacking Wallerian degeneration . DB01017 , a broad-spectrum antimicrobial tetracycline , acts neuroprotectively in ischemia . Recently , however , minocycline has been revealed to have ambiguous effects on nerve regeneration . Thus its effects in a rat sciatic nerve transplantation model and on cultivated Schwann cells stressed by oxygen glucose deprivation ( OGD ) were studied . The negative effect of minocycline on Wallerian degeneration , the essential initial phase of degeneration/regeneration after nerve injury , that was recently demonstrated , was excluded by using predegenerated nerve and Schwann cell-enriched muscle grafts , both free of Wallerian degeneration . They were compared with common nerve grafts . The principle findings were that in vitro minocycline provided protective effects against OGD-induced death of Schwann cells by preventing permeability of the mitochondrial membrane . It suppressed the OGD-mediated induction of HIF-1alpha and Q07812 , and stabilized/induced BCL-2 . P99999 release and cleavage of procaspase-3 were diminished ; release and translocation of O95831 and cytotoxic cleavage of actin into fractin were stopped . In common nerve grafts , minocycline , besides its direct anti-ischemic effect , hampered revascularization by down-regulation of P14780 and P15692 prolonging ischemia and impeding macrophage recruitment . In bioartificial nerve grafts that were free of Wallerian degeneration and revealed lower immunogenicity , minocycline aided the regeneration process . Here , the direct anti-ischemic effect of minocycline on Schwann cells , which are mandatory for successful peripheral nerve regeneration , dominated the systemic anti-angiogenic/pro-ischemic effects . In common nerve grafts , however , where Wallerian degeneration is a prerequisite , the anti-angiogenic and macrophage-depressing effect is an obstacle for regeneration . Lupus : novel therapies in clinical development . There have been significant advancements in understanding the immunopathogenesis of systemic lupus erythematosus . However , the developments in therapeutics have been rather slow . DB08879 , a Q9Y275 ( Q9Y275 ) inhibitor has been approved for the treatment of this disease after more than 50 years . Numerous biological agents are being developed which target the B cells , T cells , and various cytokines . Among anti-B cell therapy , drugs target P11836 + cells ( ocrelizumab , SBI-087 ) , P20273 + cells ( epratuzumab ) \or the receptors of tumor necrosis factor ( P01375 ) superfamily ( atacicept , LY2127399 , A-623 ) . Monoclonal antibodies targeting interferon alpha ( IFN-α ) and gamma ( IFN-γ ) and interleukins ( P05231 , 10 ) are being investigated for SLE . Novel targets include toll like receptors , phosphodiesterases , P29965 and retinoid receptors . This review discusses various drugs which are in different phases of clinical trials and hold promise for patients suffering from this chronic debilitating disease . P. aeruginosa drives P10145 synthesis via redundant toll-like receptors and NADPH oxidase in CFTR∆F508 airway epithelial cells . BACKGROUND : Understanding the mechanisms underlying bacterial-driven inflammation and neutrophil recruitment is important to design better therapies for CF . P10145 is an important chemokine found elevated in the airways of CF patients that recruits neutrophil to sites of the inflammation . METHODS : Airway epithelial cells ( AECs ) expressing wild-type P13569 or CFTR∆F508 were challenged with Pseudomonas aeruginosa diffusible material ( PsaDM ) and the synthesis of P10145 was measured by quantitative real-time PCR and ELISA in absence or presence of MAPK inhibitors , TLR antagonists , glutathione and a NADPH oxidase inhibitor . RESULTS : CFTR∆F508 AECs secrete more P10145 in response to PsaDM than their wild type counterpart , which can be reversed by addition of extracellular glutathione or incubating AECs at 27°C to favour folding and expression of P13569 at the cell membrane . Moreover , in CFTR∆F508 AECs , O60603 , O00206 and O60602 act redundantly to drive P10145 synthesis via the activation of NADPH oxidase . DISCUSSIONS : These results demonstrate that NADPH oxidase is necessary for P10145 synthesis in response to TLRs activation by P. aeruginosa . DB01017 reduces renal microvascular leakage in a rat model of ischemic renal injury . Tetracyclines exhibit significant anti-inflammatory properties , inhibit matrix metalloproteinases ( MMPs ) , and are protective in models of ischemia-reperfusion injury ( IRI ) . Both inflammatory cascades and MMP activation have been demonstrated to modulate microvascular permeability . Because increased microvascular permeability occurs during IRI in a variety of organ systems including the kidney , we hypothesized that minocycline , a semisynthetic tetracycline , would diminish microvascular leakage during renal IRI . To test this hypothesis , we used intravital 2-photon microscopy to examine leakage of fluorescent dextrans from the vasculature in a rodent model of IRI . DB01017 significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Although minocycline diminished leukocyte accumulation in the kidney following ischemia , areas of leukocyte accumulation did not correlate with areas of microvascular permeability in either the saline- or minocycline-pretreated animals . DB01017 diminished the perivascular increase in P08253 and P14780 , as well as the increase in P08253 activity 24 h after ischemia . ABT-518 , a specific inhibitor of P08253 and P14780 , also significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Our results indicate that minocycline mitigates the renal microvascular permeability defect following IRI . This effect is spatially distinct from the effect of minocycline on leukocyte accumulation and may be related to diminished activity of MMPs on the integrity of the perivascular matrix . DB01017 inhibits P09917 activation and brain inflammation after focal cerebral ischemia in rats . AIM : To determine whether the anti-inflammatory effect of minocycline on postischemic brain injury is mediated by the inhibition of P09917 ( 5- P28300 ) expression and enzymatic activation in rats . METHODS : Focal cerebral ischemia was induced for 30 min with middle cerebral artery occlusion , followed by reperfusion . The ischemic injuries , endogenous IgG exudation , the accumulation of neutrophils and macrophage/microglia , and 5- P28300 mRNA expression were determined 72 h after reperfusion . 5- P28300 metabolites ( leukotriene B4 and cysteinyl leukotrienes ) were measured 3 h after reperfusion . RESULTS : DB01017 ( 22.5 and 45 mg/kg , ip , for 3 d ) attenuated ischemic injuries , IgG exudation , and the accumulation of neutrophils and macrophage/microglia 72 h after reperfusion . It also inhibited 5- P28300 expression 72 h after reperfusion and the production of leukotrienes 3 h after reperfusion . CONCLUSION : DB01017 inhibited postischemic brain inflammation , which might be partly mediated by the inhibition of 5- P28300 expression and enzymatic activation . DB01017 inhibits malignant ascites of ovarian cancer through targeting multiple signaling pathways . OBJECTIVES : To evaluate the effect of minocycline on the expression of cytokines and growth factors responsible for malignant ascite formation . METHODS : In vitro , cells obtained from malignant ascites were pre-treated with minocycline ( 0-100 μmol/L ) and exposed briefly to hypoxia . In vivo , female nude mice bearing OVCAR-3 tumors were treated orally in drinking water with minocycline for 4 weeks . Plasma , ascites , and tumors were analyzed . RESULTS : DB01017 blocked hypoxia-induced surge in interleukin-6 ( P05231 ) , its soluble receptor ( sIL-6R ) and vascular endothelial growth factor ( P15692 ) levels in concentration-dependent manner . In mice , orally administered minocycline led to dramatic reduction in tumor weight and malignant ascite volume . P05231 , sIL6R and in particular P15692 levels were highly suppressed in plasma , ascite fluid and tumor tissue by minocycline . In addition , tumors from minocycline treated mice expressed profoundly lower levels of phosphorylated extracellular regulated kinases ( p-Erk1/2 ) and p-Akt . DB01017 was also effective at suppressing transforming growth factor beta ( TGF-β1 ) and increasing vascular endothelial cadherin ( P33151 ) expression thereby providing molecular confirmation for its effects on malignant ascite formation . CONCLUSION : Orally administered minocycline is highly effective in suppressing ovarian cancer-induced malignant ascites by targeting cytokines and growth factors essential for tumor growth and malignant ascite formation . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . DB01017 inhibits P09917 expression and accelerates functional recovery in chronic phase of focal cerebral ischemia in rats . AIMS : We previously reported that minocycline attenuates acute brain injury and inflammation after focal cerebral ischemia , and this is partly mediated by inhibition of P09917 ( 5- P28300 ) expression . Here , we determined the protective effect of minocycline on chronic ischemic brain injury and its relation with the inhibition of 5- P28300 expression after focal cerebral ischemia . MAIN METHODS : Focal cerebral ischemia was induced by 90 min of middle cerebral artery occlusion followed by reperfusion for 36 days . DB01017 ( 45 mg/kg ) was administered intraperitoneally 2h and 12h after ischemia and then every 12h for 5 days . Sensorimotor function was evaluated 1-28 days after ischemia and cognitive function was determined 30-35 days after ischemia . Thereafter , infarct volume , neuron density , astrogliosis , and 5- P28300 expression in the brain were determined . KEY FINDINGS : DB01017 accelerated the recovery of sensorimotor and cognitive functions , attenuated the loss of neuron density , and inhibited astrogliosis in the boundary zone around the ischemic core , but did not affect infarct volume . DB01017 significantly inhibited the increased 5- P28300 expression in the proliferated astrocytes in the boundary zone , and in the macrophages/microglia in the ischemic core . SIGNIFICANCE : DB01017 accelerates functional recovery in the chronic phase of focal cerebral ischemia , which may be partly associated with the reduction of 5- P28300 expression . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Toll-like receptor expression in human keratinocytes : nuclear factor kappaB controlled gene activation by Staphylococcus aureus is toll-like receptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent . Cultured primary human keratinocytes were screened for their expression of various members of the toll-like receptor ( TLR ) family . Keratinocytes were found to constitutively express Q15399 , O60603 , O15455 , O60602 , and Q9NR96 but not O00206 , Q9Y2C9 , Q9NYK1 , Q9NR97 , or Q9BXR5 as shown by polymerase chain reaction analysis . The expression of the crucial receptor for signaling of staphylococcal compounds O60603 was also confirmed by immunohistochemistry , in contrast to O00206 , which showed a negative staining pattern . Next , we analyzed the activation of the proinflammatory nuclear transcription factor kappaB by Staphylococcus aureus strain 8325-4 . Using nuclear extract gel shifts , RelA staining , and luciferase reporter transfection plasmids we found a clear induction of nuclear factor kappaB translocation by the bacteria . This translocation induced the transcription of nuclear factor kappaB controlled genes such as inducible nitric oxide synthetase , P35354 , and interleukin-8 . Transcription of these genes was followed by production of increased amounts of interleukin-8 protein and NO . Inhibition experiments using monoclonal antibodies and the specific platelet activating factor receptor inhibitor CV3988 showed that nuclear factor kappaB activation by S. aureus was O60603 but not O00206 or platelet activating factor receptor dependent . In line , the purified staphylococcal cell wall components lipoteichoic acid and peptidoglycan , known to signal through O60603 , also showed nuclear factor kappaB translocation in human keratinocytes , indicating a crucial role of the staphylococcal cell wall in the innate immune stimulation of human keratinocytes . These results help to explain the complex activation of human keratinocytes by S. aureus and its cell wall components in various inflammatory disorders of the skin . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Understanding the molecular mechanism of blood-brain barrier damage in an experimental model of Japanese encephalitis : correlation with minocycline administration as a therapeutic agent . The blood-brain barrier ( BBB ) serves to protect the central nervous system ( CNS ) from damage by exogenous molecules . Japanese encephalitis ( JE ) , caused by a neurotropic flavivirus , leads to inflammation in the CNS , neuronal death and also compromises the structural and functional integrity of the BBB . DB01017 , a semisynthetic tetracycline , has been found to be broadly protective in neurological disease models featuring inflammation and cell death and at present , is being evaluated in clinical trials . In the present study , we propose that the neuroprotective role of minocycline in experimental models of JE extends also to the protection of the BBB . Damage to the BBB was assessed by Evan 's blue dye exclusion test after minocycline treatment following Japanese encephalitis virus ( JEV ) infection . A breakdown of the BBB occurred in mice inoculated intravenously with JEV . This resulted in leakage of protein-bound Evan 's blue dye into the brain tissue . Semi-quantitative RT-PCR revealed an up-regulation of chemokine receptors and adhesion molecules following JEV infection . Immunostaining showed leukocyte and neutrophil infiltration following JEV infection . Intraperitoneal injection of minocycline , beginning 24h post-JEV infection , abrogated the effects by reducing BBB damage , decreasing expression of P35228 , Cox-2 , P15692 and also by reducing the elevated level of transcript of chemokine receptors and adhesion molecules in the brain . Matrix metalloproteinases ( MMPs ) are known to disrupt the BBB and minocycline was found to significantly decrease the activity of P14780 in brain tissue homogenates . Thus , minocycline , administered at a clinically relevant time , appears to maintain blood-brain barrier integrity following JEV infection . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Eicosanoid signalling pathways in the heart . Myocardial phospholipids serve as primary reservoirs of arachidonic acid ( AA ) , which is liberated through the rate-determining hydrolytic action of cardiac phospholipases A2 ( PLA2s ) . A predominant P04054 in myocardium is calcium-independent phospholipase A2beta ( iPLA2beta ) , which , through its calmodulin ( P62158 ) and DB00171 -binding domains , is regulated by alterations in local cellular Ca2+ concentrations and cardiac bioenergetic status , respectively . Importantly , iPLA2beta has been demonstrated to be activated by ischaemia through elevation of the concentration of myocardial fatty acyl- DB01992 , which abrogates Ca2+/ P62158 -mediated inhibition of iPLA2beta . AA released by P04054 -catalysed hydrolysis of phospholipids serves as a precursor for eicosanoids generated by pathways dependent on cyclooxygenases ( P36551 ) , lipoxygenases ( P28300 ) , and cytochromes P450 ( CYP ) . Eicosanoids initiate and propagate diverse signalling cascades , primarily through their interaction with cellular receptors and ion channels . However , during pathologic states such as ischaemia or congestive heart failure , eicosanoids contribute to multiple maladaptive changes including inflammation , alterations of cellular growth programmes , and activation of multiple transcriptional events leading to the deleterious sequelae of these pathologic states . This review summarizes the central roles of myocardial PLA(2)s in eicosanoid signalling in the heart , the major P36551 , P28300 , and CYP pathways of eicosanoid generation in the myocardium , and the effects of important eicosanoids on receptor- , ion channel- , and transcription-mediated processes that facilitate cardiac hypertrophy , mediate ischaemic preconditioning , and precipitate arrhythmogenesis in response to pathologic stimuli . Glioma-associated microglial P14780 expression is upregulated by O60603 signaling and sensitive to minocycline . The invasiveness of malignant gliomas is one of the major obstacles in glioma therapy and the reason for the poor survival of patients . Glioma cells infiltrate into the brain parenchyma and thereby escape surgical resection . Glioma associated microglia/macrophages support glioma infiltration into the brain parenchyma by increased expression and activation of extracellular matrix degrading proteases such as matrix metalloprotease ( MMP ) 2 , P14780 and membrane-type 1 MMP . In this work we demonstrate that , P14780 is predominantly expressed by glioma associated microglia/macrophages in mouse and human glioma tissue but not by the glioma cells . Supernatant from glioma cells induced the expression of P14780 in cultured microglial cells . Using mice deficient for different Toll-like receptors we identified O60603 /6 as the signaling pathway for the glioma induced upregulation of microglial P14780 . Also in an experimental mouse glioma model , O60603 deficiency attenuated the upregulation of microglial P14780 . Moreover , glioma supernatant triggered an upregulation of O60603 expression in microglia . Both , the upregulation of P14780 and O60603 were attenuated by the antibiotic minocycline and a p38 mitogen-activated protein kinase antagonist in vitro . DB01017 also extended the survival rate of glioma bearing mice when given to the drinking water . Thus glioma cells change the phenotype of glioma associated microglia/macrophages in a complex fashion using O60603 as an important signaling pathway and minocycline further proved to be a potential candidate for adjuvant glioma therapy . Benzyl isothiocyanate ( BITC ) inhibits migration and invasion of human gastric cancer AGS cells via suppressing P29323 signal pathways . Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases . Benzyl isothiocyanate ( BITC ) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables . BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans . Here , the purpose of this study was to investigate the effects of BITC on cell proliferation , migration , invasion and mitogen-activated protein kinase ( MAPK ) pathways of AGS human gastric cancer cells . Wound healing and Boyden chamber ( migration and invasion ) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells . BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner . Results from Western blotting indicated that BITC exerted an inhibitory effect on the P27361 /2 , Ras , P62993 , Rho A , P35228 , P35354 for causing the inhibitions of P08253 , -7 and -9 then followed by the inhibitions of invasion and migration of AGS cells in vitro . BITC also promoted O14733 , Q99759 , c-jun , P45983 /2 , P15692 , Sos1 , phosphoinositide 3-kinase ( PI3K ) , PKC , nuclear factor-kappaB ( NF-κB ) p65 in AGS cells . Results from real-time polymerized chain reaction ( PCR ) showed that BITC inhibited the gene expressions of P08253 ,-7 -9 , Q05397 , Q13464 and RhoA after BITC treatment for 24 and 48 hours in AGS cells . Taken together , the finding may provide new mechanisms and functions of BITC , which inhibit migration and invasion of human gastric cancer AGS cells . In vitro effect of thymosin-alpha1 and interferon-alpha on Th1 and Th2 cytokine synthesis in patients with chronic hepatitis C . Current evidence suggests that increased expression of Th1-associated cytokines is important for immune-mediated eradication of hepatitis C infection , while an increase in Th2-associated cytokines is associated with persistence of infection . In this study we evaluated the effects of thymosin-alpha1 ( Q96RJ0 ) , a naturally occurring thymic peptide , and interferon-alpha ( IFN-alpha ) on cytokine production in peripheral blood mononuclear cells from untreated patients with chronic hepatitis C . We examined the effect of incubation with Q96RJ0 , IFN-alpha , or both , on production of Th1-associated cytokines ( P60568 , P01579 ) , Th2-associated cytokines ( P05112 , P22301 ) , and synthesis of the antiviral protein 2',5'-oligoadenylate synthetase . Q96RJ0 treatment induced a significant increase in production of P60568 and 2',5'-oligoadenylate synthetase . Smaller increases were also seen after treatment with IFN-alpha , while incubation with Q96RJ0 and IFN-alpha together led to an additive or synergistic effect . Incubation with Q96RJ0 resulted in a decrease in P05112 and P22301 , whereas IFN-alpha increased these cytokines . The addition of Q96RJ0 to IFN-alpha significantly reversed this IFN-alpha-induced increase . Hence , Q96RJ0 treatment could benefit patients with hepatitis C infection by increasing the Th1-type response , fundamental for sustained clearance of hepatitis C ; and by decreasing the Th2-type response , associated with persistence of viraemia . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . Protection of minocycline on early brain injury after subarachnoid hemorrhage in rats . DB01017 has been shown to be neuroprotective in cerebral ischemia and in other models of brain injury . Our goal is to observe the protection of minocycline on EBI after Q53FZ2 and the mechanism . 48 adult male SD rats were randomly divided into four groups : the sham-operated group , Q53FZ2 group , vehicle group ( Q53FZ2 +normal sodium ) , and minocycline group ( Q53FZ2 +minocycline ) . The Q53FZ2 model was induced by injecting 300 μl of autologous arterial blood into the prechiasmatic cistern . Expressions of P14780 in the hippocampus were examined at 24 h by western blot and zymography . Western blot and zymography showed that the expression of total and active P14780 increased dramatically at 24 h after Q53FZ2 compared with that of the sham group ( P < 0.01 ) . The clinical assessments got a lower score than that of the sham-operated group . After treated with minocycline , the expression of P14780 decreased significantly ( P < 0.01 vs. vehicle group ) , and the clinical assessments improved . We conclude that minocycline can protect EBI after Q53FZ2 , which may be related to the mechanism of inhibiting the expression of P14780 in the hippocampus . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . Donor pre-treatment with everolimus or cyclosporine does not reduce ischaemia-reperfusion injury in a rat kidney transplant model . BACKGROUND : Immunosuppressive agents have been investigated in renal ischaemia-reperfusion injury ( IRI ) and have frequently demonstrated a beneficial effect . Most studies focused on treatment of the recipient at the time of transplantation . Pre-treatment of these organs before injury ( pharmacological pre-conditioning ) may particularly protect these organs . This study aimed to investigate the possible protective effects of donor pre-treatment with cyclosporine ( DB00091 ) or the P42345 inhibitor everolimus or their combination against IRI during renal transplantation in a rat model . METHODS : Donors received vehicle , DB00091 ( 5 mg/kg ) , everolimus ( 0.5 mg/kg ) or CsA + everolimus . Two oral doses were administered to the donors at 24 h and again at 6 h prior to donor kidney removal . Syngeneic rat kidneys were preserved in UW solution for 24 h prior to transplantation . After 24 h of reperfusion , blood and tissue samples were collected from recipients for further analysis . RESULTS : Renal functions as determined by creatinine and necrosis scores were not different between the experimental groups . Cleaved caspase-3 , heat shock protein 70 ( HSP70 ) , tumor-necrosis factor-alpha ( P01375 -α ) and nitrotyrosine protein levels were not statistically different between the four treatment groups at 24 h post-transplantation . Blood NMR analysis on metabolic markers for IRI reveals no beneficial effects of donor pre-treatment on the 24-h outcome in transplantation . CONCLUSIONS : When given alone or as a combination to donors before organ recovery , cyclosporine or everolimus does not appear to ameliorate IRI . P10451 increases migration and P14780 up-regulation via alphavbeta3 integrin , Q05397 , P29323 , and NF-kappaB-dependent pathway in human chondrosarcoma cells . Tumor malignancy is associated with several features such as proliferation ability and frequency of metastasis . P10451 ( P10451 ) , which abundantly expressed in bone matrix , is involved in cell adhesion , migration , invasion and proliferation via interaction with its receptor , that is , alphavbeta3 integrin . However , the effect of P10451 on migration activity in human chondrosarcoma cells is mostly unknown . Here we found that P10451 increased the migration and expression of matrix metalloproteinase ( MMP ) -9 in human chondrosarcoma cells ( JJ012 cells ) . RGD peptide , alphavbeta3 monoclonal antibody and MAPK kinase ( MEK ) inhibitors ( PD98059 and U0126 ) but not P55042 peptide inhibited the P10451 -induced increase of the migration and P14780 up-regulation of chondrosarcoma cells . P10451 stimulation increased the phosphorylation of focal adhesion kinase ( Q05397 ) , MEK and extracellular signal-regulated kinase ( P29323 ) . In addition , treatment of JJ012 cells with NF-kappaB inhibitor ( PDTC ) or IkappaB protease inhibitor ( TPCK ) inhibited P10451 -induced cell migration and P14780 up-regulation . Stimulation of JJ012 cells with P10451 also induced O15111 alpha/beta ( IKK alpha/beta ) phosphorylation , P25963 phosphorylation , p65 DB00133 (536) phosphorylation , and kappaB-luciferase activity . The P10451 -mediated increases in P14780 and kappaB-luciferase activities were inhibited by RGD peptide , PD98059 or Q05397 and P28482 mutant . Taken together , our results indicated that P10451 enhances the migration of chondrosarcoma cells by increasing P14780 expression through the alphavbeta3 integrin , Q05397 , MEK , P29323 and NF-kappaB signal transduction pathway . Ds-echinoside A , a new triterpene glycoside derived from sea cucumber , exhibits antimetastatic activity via the inhibition of NF-κB-dependent P14780 and P15692 expressions . Ds-echinoside A ( DSEA ) , a non-sulfated triterpene glycoside , was isolated from the sea cucumber Pearsonothuria graeffei . In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion , migration , invasion , and angiogenesis . In this study , we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2 , with a half-maximal inhibitory concentration ( IC₅₀ ) of 2.65 μmol/L , and suppressed Hep G2 cell adhesion , migration , and invasion in a dose-dependent manner . DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane ( P62158 ) assay in vivo . Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 ( P14780 ) , which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis . DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 ( P01033 ) , an important regulator of P14780 activation . From the results of Western blotting , the expressions of nuclear factor-kappa B ( NF-κB ) and vascular endothelial growth factor ( P15692 ) were found to be remarkably reduced by DSEA . These findings suggest that DSEA exhibits a significant anti-metastatic activity through the specific inhibition of NF-κB-dependent P14780 and P15692 expressions . P14780 in an exploratory trial of intravenous minocycline for acute ischemic stroke . BACKGROUND AND PURPOSE : Plasma matrix metalloproteinase-9 levels predict posttissue plasminogen activator ( tPA ) hemorrhage . METHODS : The authors investigated the effect of minocycline on plasma matrix metalloproteinase-9 in acute ischemic stroke in the DB01017 to Improve Neurological Outcome in Stroke ( MINOS ) trial and a comparison group . RESULTS : P14780 level decreased at 72 hours compared with baseline in MINOS ( tPA , P=0.0022 ; non-tPA , P=0.0066 ) and was lower than in the non-MINOS comparison group at 24 hours ( tPA , P < 0.0001 ; non-tPA , P=0.0019 ) . CONCLUSIONS : Lower plasma matrix metalloproteinase-9 was seen among tPA-treated subjects in the MINOS trial . Combining minocycline with tPA may prevent the adverse consequences of thrombolytic therapy through suppression of matrix metalloproteinase-9 activity . Differential induction of matrix metalloproteinase 1 and 2 in ectopic endometrium . According to the transplantation theory , endometriosis develops from endometrial fragments that are retrogradely menstruated into the peritoneal cavity . In order to develop into endometriotic lesions , they have to connect to the vascular system by angiogenesis , probably involving matrix metalloproteinases ( MMP ) as key enzymes in extracellular matrix remodelling . A model of endometriosis using the chorioallantoic membrane ( P62158 ) of chick embryos was established . Eutopic endometrium from healthy women was transferred to the P62158 and cultivated ectopically for up to 3 days . Before transplantation and after 24 , 48 and 72 h of culture on the P62158 , total RNA was extracted and reverse transcribed . Human P03956 ( interstitial collagenase ) and P08253 ( gelatinase A ) mRNA expression was assessed by competitive PCR . Results were normalized to the content of human glyceraldehyde 3-phosphate dehydrogenase ( P04406 ) mRNA . In eutopic endometrium , 0.29 amol P03956 mRNA and 0.42 fmol P08253 mRNA per fmol P04406 mRNA were found . Relative P03956 mRNA concentrations increased strongly after culture on the P62158 , while P08253 mRNA levels were nearly unaltered . This differential regulation suggests different roles of these enzymes in the angiogenesis of ectopic endometrial fragments and during the development of endometriosis . DB00640 regulation of cystic fibrosis transmembrane conductance regulator through prostenoids in airway epithelia . Cystic fibrosis is caused by dysfunction of the cystic fibrosis transmembrane conductance regulator ( P13569 ) protein , leading to altered ion transport , chronic infection , and excessive inflammation . Here we investigated regulation of P13569 in airway cell monolayers by adenosine , adenosine receptors , and arachidonic acid . Our studies demonstrate that the A2B adenosine receptor is expressed at high levels relative to the other adenosine receptor subtypes , with a characteristic low-affinity profile for adenosine-stimulated P13569 Cl- currents in both Calu-3 cells and CFBE41o- airway cell monolayers stably transduced with wild-type P13569 . The levels of adenosine found in sputum from patients with cystic fibrosis with moderate to severe lung disease stimulated apical prostaglandin release in Calu-3 and CFBE41o- cells , implicating adenosine regulation of phospholipase A2 ( P04054 ) activity . A2B adenosine receptor and arachidonic acid stimulation produced P13569 -dependent currents in airway monolayers and increased DB02527 levels that were sensitive to cyclooxygenase inhibition . Arachidonic acid demonstrated dual regulation of P13569 , stimulating P13569 and Cl- currents in intact airway monolayers , and potently inhibiting PKA-activated Cl- currents in excised membrane patches . Cl- currents produced by arachidonic acid were sensitive to inhibition of PKA , cyclooxygenase , and P09917 . Together , the results provide a converging mechanism to link regulation of P13569 and airway cell inflammation through adenosine and adenosine receptors . B lymphocytes and B-cell activating factor promote collagen and profibrotic markers expression by dermal fibroblasts in systemic sclerosis . INTRODUCTION : B lymphocytes might play a pathogenic role in dermal fibrosis in systemic sclerosis ( SSc ) . B-cell activating factor ( Q9Y275 ) , a key cytokine for B-cell activation , is increased in the serum and the skin of patients with SSc . However , the ability of B cells directly to stimulate dermal fibroblasts and the role of Q9Y275 are not fully understood . We therefore investigated the involvement of B cells and Q9Y275 in the expression of collagen and profibrotic markers by dermal fibroblasts . METHODS : Cocultures of blood B cells from healthy blood donors and normal or SSc dermal fibroblasts stimulated with anti-IgM and Q9Y275 were performed . Alpha-SMA , P01033 , P14780 , P02452 , P08123 , and P02461 mRNA expression were determined by quantitative RT-PCR . Soluble collagen , Q9Y275 , P05231 , IL-1β , TGF-β1 , and P13500 protein secretion were assessed . RESULTS : Coculture of blood B cells and dermal fibroblasts isolated from SSc patients induced P05231 , TGF-β1 , P13500 , and collagen secretion , as well as Alpha-SMA , P01033 , and P14780 expression in dermal fibroblasts . Transwell assays demonstrated that this induction was dependent on cell-cell contact . Addition of anti-IgM and Q9Y275 to the coculture increased P05231 , P13500 , TGF-β1 , and collagen secretion . B cell- and Q9Y275 -induced collagen secretion was highly reduced by anti-TGF-β1 antibodies . CONCLUSIONS : Our results showed for the first time a direct role of B cells on the production of collagen by dermal fibroblasts , which is further enhanced by Q9Y275 . Thus , these results demonstrate a new pathogenic role of B cells and Q9Y275 in fibrosis and systemic sclerosis . DB01017 protects against permanent cerebral ischemia in wild type but not in matrix metalloprotease-9-deficient mice . DB01017 is protective in models of transient middle cerebral artery occlusion ( MCAO ) . We studied whether minocycline and doxycycline , another tetracycline derivative , provide protection in permanent MCAO . Because minocycline inhibits matrix metalloprotease-9 ( P14780 ) , we also compared minocycline 's protective effect in wild type ( wt ) and P14780 knock-out ( ko ) mice . Wt FVB/N , Balb/C , and two lines of P14780 ko and their wt C57Bl/6 control mice were subjected to 24- or 72-hour permanent MCAO . Drug administration was started either 12 hours before or 2 hours after the onset of MCAO . Infarct size was determined by triphenyltetrazolium staining or P24752 -weighted Q9BWK5 . Zymography was used to study the expression of MMPs . In wt strains , tetracycline treatments started before MCAO reduced the infarct size by 25 % to 50 % , whereas the treatment started after MCAO was not protective . DB01017 inhibited ischemia-provoked pro- P14780 induction in wt mice , but was not protective in P14780 ko mice . Pro- P08253 was induced by MCAO in wt and P14780 ko mice . MCAO-induced pro- P08253 was downregulated by minocycline treatment in wt mice but remained in P14780 ko mice at the same level as in saline-treated wt mice . Tetracyclines are protective in permanent MCAO when the treatment is started before the insult . DB01017 may provide protection by interfering with MMPs . P01375 -α-accelerated degradation of type I collagen in human skin is associated with elevated matrix metalloproteinase ( MMP ) -1 and P08254 ex vivo . P01375 ( P01375 ) -α induces matrix metalloproteinases ( MMPs ) that may disrupt skin integrity . We have investigated the effects and mechanisms of exogenous P01375 -α on collagen degradation by incubating human skin explants in defined serum-free media with or without P01375 -α ( 10ng/ml ) in the absence or presence of the nonselective MMP inhibitor DB02255 for 8 days . The basal culture conditions promoted type I collagen catabolism that was accelerated by P01375 -α ( p < 0.005 ) and accomplished by MMPs ( p < 0.005 ) . Levels of the collagenases P22894 and P45452 were insignificant and neither P08253 nor P50281 were associated with increased collagen degradation . P01375 -α increased secretion of P03956 ( p < 0.01 ) but had no impact on P03956 quantities in the tissue . Immunohistochemical analysis confirmed similar tissue P03956 expression with or without P01375 -α with epidermis being the major source of P03956 . Increased tissue-derived collagenolytic activity with P01375 -α exposure was blocked by neutralizing P03956 monoclonal antibody and was not due to down-regulation of tissue inhibitor of metalloproteinase-1 . P01375 -α increased production ( p < 0.01 ) , tissue levels ( p < 0.005 ) and catalytic activity of the endogenous P03956 activator P08254 . Type I collagen degradation correlated with P08254 tissue levels ( rs=0.68 , p < 0.05 ) and was attenuated with selective P08254 inhibitor . Type I collagen formation was down-regulated in cultured compared with native skin explants but was not reduced further by P01375 -α . P01375 -α had no significant effect on epidermal apoptosis . Our data indicate that P01375 -α augments collagenolytic activity of P03956 , possibly through up-regulation of P08254 leading to gradual loss of type I collagen in human skin . Characterization of a hydroperoxide lyase gene and effect of P13671 -volatiles on expression of genes of the oxylipin metabolism in Citrus . A number of P13671 -volatile products of the lipoxygenase ( P28300 ) pathway was examined for their antifungal activity and a potential role as a signal molecule in citrus . trans-2-Hexenal induced the rough lemon lipoxygenase gene ( RlemLOX ) , hydroperoxide lyase gene ( RlemHPL ) and AOS gene , but hexanal , and hexanol suppressed them . cis-3-Hexenol and trans-2-hexenol increased expression of the AOS gene but not RlemLOX and RlemHPL . Transcripts of the RlemHPL and AOS gene were detected constitutively in leaves by northern blot , but wounding or inoculation with nonpathogenic Alternaria alternata rapidly increased the transcript accumulation . Transcripts of the RlemHPL and AOS genes were also induced with pathogenic A. alternata , which produces the host-selective P10323 -toxin , but the signal declined rapidly after inoculation . An increase in enzymatic activity of P09382 after wounding or inoculation with nonpathogen was suppressed in leaves infected with the pathogen . Interestingly , vapor treatment with trans-2-hexenol delayed necrotic spot formation in the leaves inoculated with the pathogenic A. alternata . Since trans-2-hexenol has no antifungal activity to A. alternata and also did not inhibit necrosis formation by P10323 -toxin alone , the delay of symptoms may be caused by activation of AOS in the P28300 pathway to produce oxylipin derivatives such as methyl jasmonate for activation of defense related genes with antifungal activity . Oxidative stress induces extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase in cystic fibrosis lung epithelial cells : Potential mechanism for excessive P10145 expression . Cystic fibrosis ( CF ) is a lethal disease caused by defective function of the cftr gene product , the CF transmembrane conductance regulator ( P13569 ) that leads to oxidative damage and excessive inflammatory response in lungs of CF patients . We here report the effects of oxidative stress ( hyperoxia , 95 % O(2) ) on the expression of pro-inflammatory interleukin ( IL ) -8 and P25024 /2 receptors in two human CF lung epithelial cell lines ( IB3-1 , with the heterozygous F508del/W1282X mutation and CFBE41o- with the homozygous F508del/F508del mutation ) and two control non-CF lung epithelial cell lines ( S9 cell line derived from IB3-1 after correction with wtCFTR and the normal bronchial cell line 16HBE14o- ) . Under oxidative stress , the expression of P10145 and P25024 /2 receptors was increased in CF , corrected and normal lung cell lines . The effects of oxidative stress were also investigated by measuring the transcription nuclear factor kappaB ( NF-kappaB ) and activator protein-1 ( AP-1 ) activities . Under oxidative stress , no increase of NF-kappaB activation was observed in CF lung cells in contrast to that observed in normal and corrected CF lung cells . The signalling of mitogen-activated protein ( Q96HU1 ) kinases was further studied . We demonstrated that extracellular signal-regulated kinase ( P27361 /2 ) and AP-1 activity was markedly enhanced in CF but not non-CF lung cells under oxidative stress . Consistently , inhibition of P27361 /2 in oxidative stress-exposed CF lung cells strongly decreased both the P10145 production and P25024 /2 expression . Therefore , targeting of P27361 /2 Q96HU1 kinase may be critical to reduce oxidative stress-mediated inflammation in lungs of CF patients . Involvement of pro- and antinociceptive factors in minocycline analgesia in rat neuropathic pain model . In neuropathic pain the repeated minocycline treatment inhibited the mRNA and protein expression of the microglial markers and metalloproteinase-9 ( P14780 ) . The minocycline diminished the pronociceptive ( P05231 , Q14116 ) , but not antinociceptive ( IL-1alpha , P05112 , P22301 ) cytokines at the spinal cord level . In vitro primary cell culture studies have shown that P14780 , P01033 , IL-1beta , IL-1alpha , P05231 , P22301 , and Q14116 are of microglial origin . DB01017 reduces the production of pronociceptive factors , resulting in a more potent antinociceptive effect . This change in the ratio between pro- and antinociceptive factors , in favour of the latter may be the mechanism of minocycline analgesia in neuropathy . DB01017 accelerates hypoxia-inducible factor-1 alpha degradation and inhibits hypoxia-induced neovasculogenesis through prolyl hydroxylase , von Hippel-Lindau-dependent pathway . Hypoxia-mediated stress responses are important in tumor progression , especially when tumor growth causes the tumor to become deprived of its blood supply . The oxygen-labile transcription factor hypoxia-inducible factor-1 alpha ( HIF-1α ) plays a critical role in regulating hypoxia stress-related gene expression and is considered a novel therapeutic target . Lung adenocarcinoma cell lines were exposed to minocycline , followed by incubation at hypoxic condition for 3-6 h . Here , we show that minocycline , a second-generation tetracycline , can induce HIF-1α proteasomal degradation under hypoxia by increasing the expression prolyl hydroxylase-2 and HIF-1α/von Hippel-Lindau protein interaction , thereby overcoming hypoxia-induced HIF-1α stabilization . Neither repression of hypoxia-induced phosphatidylinositol-3 kinase/Akt/mammalian target of rapamycin pathway nor inhibition of Hsp90 was required for minocycline-induced HIF-1α degradation . The HIF-1α degradation-enhancing effect of minocycline was evident in both cancerous and primary cells . DB01017 -pretreated , hypoxia-conditioned cells showed a clear reduction in hypoxia response element reporter expression and amelioration of vascular endothelial growth factor C/D ( P49767 /D ) , matrix metalloproteinase 2 , and glucose transporter 1 expression . By decreasing P15692 secretion of cancerous cells , minocycline could suppress endothelial cell neovasculogenesis . These findings suggest a novel application of minocycline in the treatment of tumor angiogenesis as well as hypoxia-related diseases . Depletion of Nrf2 enhances inflammation induced by oxyhemoglobin in cultured mice astrocytes . Q16236 ( Nrf2 ) -antioxidant response element pathway has been proved to be the key regulator in reducing inflammatory damage , which is involved in subarachnoid hemorrhage ( Q53FZ2 ) . Here , in a traditional in vitro Q53FZ2 model , we investigated the effect of Nrf2 depletion on pro-inflammatory cytokines production . Primary cultured astrocytes from Nrf2 wild type ( WT ) or knockout ( KO ) mouse were exposed or not exposed to oxyhemoglobin ( OxyHb ) . Then the DNA-binding activity of transcription factor nuclear factor-κB ( NF-κB ) was detected by EMSA . The expression of P01375 -α , IL-1β , P05231 and P14780 were evaluated . The activity of P14780 was measured by Gelatin zymography . After exposure to OxyHb , NF-κB was activated and the expression of downstream pro-inflammatory cytokines was up-regulated in astrocytes . And such up-regulation was much higher in KO astrocytes than in WT astrocytes , which means more aggravated inflammation in Nrf2 deficient astrocytes . These results suggest that astrocytes participate in inflammatory process after Q53FZ2 and the absence of Nrf2 may induce more aggressive inflammation through activation of NF-κB pathway . Serum amyloid A activates the Q96P20 inflammasome via Q99572 receptor and a cathepsin B-sensitive pathway . Serum amyloid A ( P0DJI8 ) is an acute-phase protein , the serum levels of which can increase up to 1000-fold during inflammation . P0DJI8 has a pathogenic role in amyloid A-type amyloidosis , and increased serum levels of P0DJI8 correlate with the risk for cardiovascular diseases . IL-1β is a key proinflammatory cytokine , and its secretion is strictly controlled by the inflammasomes . We studied the role of P0DJI8 in the regulation of IL-1β production and activation of the inflammasome cascade in human and mouse macrophages , as well as in THP-1 cells . P0DJI8 could provide a signal for the induction of pro-IL-1β expression and for inflammasome activation , resulting in secretion of mature IL-1β . Blocking O60603 and O00206 attenuated P0DJI8 -induced expression of P01584 , whereas inhibition of caspase-1 and the DB00171 receptor P2X(7) abrogated the release of mature IL-1β . Q96P20 inflammasome consists of the Q96P20 receptor and the adaptor protein apoptosis-associated speck-like protein containing CARD ( a caspase-recruitment domain ) ( ASC ) . P0DJI8 -mediated IL-1β secretion was markedly reduced in ASC(-/-) macrophages , and silencing Q96P20 decreased IL-1β secretion , confirming Q96P20 as the P0DJI8 -responsive inflammasome . Inflammasome activation was dependent on cathepsin B activity , but it was not associated with lysosomal destabilization . P0DJI8 also induced secretion of cathepsin B and ASC . In conclusion , P0DJI8 can induce the expression of pro-IL-1β and activation of the Q96P20 inflammasome via P2X(7) receptor and a cathepsin B-sensitive pathway . Thus , during systemic inflammation , P0DJI8 may promote the production of IL-1β in tissues . Furthermore , the P0DJI8 -induced secretion of active cathepsin B may lead to extracellular processing of P0DJI8 and , thus , potentially to the development of amyloid A amyloidosis . P04150 -mediated regulation of P14780 gene expression in human ovarian surface epithelial cells . OBJECTIVE : To obtain proof-of-concept that locally produced anti-inflammatory steroids suppress ovulation-associated extracellular matrix proteases in human ovarian surface epithelial ( OSE ) cells . DESIGN : Primary OSE cell cultures treated with interleukin-1alpha ( IL-1alpha ) ( 500 pg/mL ) as proxy for inflammation , with/without anti-inflammatory steroid ( cortisol or progesterone [ P ] , 0.01-1.0 microM ) . SETTING : Academic medical center . PATIENT(S) : Sixteen premenopausal women ( 29-46 years ) undergoing surgery for nonmalignant gynecological conditions . MAIN OUTCOME MEASURE(S) : Semiquantitative extracellular matrix protease gene expression profiling with verification by real-time quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and gelatinase zymography . RESULT(S) : Treatment with IL-1alpha stimulated messenger RNA ( mRNA ) expression of several ovulation-associated matrix metalloproteinase genes by OSE cell cultures , including gelatinase B ( P14780 ) but not gelatinase A ( P08253 ) . The IL-1alpha-stimulated P14780 mRNA production was suppressed by cortisol but not P. DB00741 but not P also dose-dependently suppressed IL-1alpha-stimulated P14780 gelatinase activity and this effect was blocked by the glucocorticoid receptor antagonist DB00834 . CONCLUSION(S) : In human OSE cells , stimulation of P14780 gene expression and proteolytic activity by IL-1alpha is suppressed by anti-inflammatory cortisol through a glucocorticoid receptor-mediated mechanism . Because IL-1alpha also generates cortisol formation in OSE by stimulating cortisone reductase activity , these results support a role for intracrine cortisol in minimizing proteolytic damage to the OSE at ovulation . Involvement of matrix metalloproteinases-2 and -9 in the formation of a lacuna-like cerebral cavity . We used a modified pial vessel disruption ( PVD ) protocol with adult male Wistar rats to mimic small-vessel stroke in the cerebral cortex . Within 3 weeks , this lesion develops into a single lacuna-like cavity , which is fluid-filled and encapsulated by reactive astrocytes . DB01017 treatment that commences 1 hr after lesion and continues for 6 days prevents the cavitation and causes a filling of the lesion with reactive astrocytes and no barrier . Here , we determined whether inhibition of matrix metalloproteinases-2 and -9 ( MMPs ) mediates this minocycline action . Confocal microscopy revealed increased punctate staining of MMPs inside the lesion sites after 2 days of PVD . Astrocytes lined the lesion border but showed sparse localization inside the lesion . In contrast , increased MMP levels inside the lesion coincided with increased Q92838 or OX-42 immunostaining , suggesting that MMP elevation reflected increased secretions from microglia/macrophages . Imaging analyses also revealed that minocycline administered for 2 days before animal euthanasia , significantly decreased MMP levels within the lesion . Moreover , Western blot analysis of cortical tissue extracts showed a significant 30-40 % upregulation of MMPs 2 days after lesion . DB01017 administered 2 hr before the lesion significantly inhibited both P14780 and P08253 levels by ∼40 % . In contrast , minocycline administered 1 hr after the lesion only decreased P14780 levels by ∼30 % . Because MMP inhibition with batimastat injection also prevented cavity formation at 21 days , we conclude that minocycline prevented the creation of a lacuna-like cyst in the cerebral cortex by inhibiting the MMP secretion from microglia in the affected tissue . A transgenic platform for testing drugs intended for reversal of cardiac remodeling identifies a novel 11βHSD1 inhibitor rescuing hypertrophy independently of re-vascularization . RATIONALE : Rescuing adverse myocardial remodeling is an unmet clinical goal and , correspondingly , pharmacological means for its intended reversal are urgently needed . OBJECTIVES : To harness a newly-developed experimental model recapitulating progressive heart failure development for the discovery of new drugs capable of reversing adverse remodeling . METHODS AND RESULTS : A P15692 -based conditional transgenic system was employed in which an induced perfusion deficit and a resultant compromised cardiac function lead to progressive remodeling and eventually heart failure . Ability of candidate drugs administered at sequential remodeling stages to reverse hypertrophy , enlarged LV size and improve cardiac function was monitored . Arguing for clinical relevance of the experimental system , clinically-used drugs operating on the P00797 -Angiotensin- DB04630 -System ( RAAS ) , namely , the P12821 inhibitor Enalapril and the direct renin inhibitor Aliskerin fully reversed remodeling . Remodeling reversal by these drugs was not accompanied by neovascularization and reached a point-of-no-return . Similarly , the PPARγ agonist Pioglitazone was proven capable of reversing all aspects of cardiac remodeling without affecting the vasculature . Extending the arsenal of remodeling-reversing drugs to pathways other than RAAS , a specific inhibitor of 11β-hydroxy-steroid dehydrogenase type 1 ( 11β HSD1 ) , a key enzyme required for generating active glucocorticoids , fully rescued myocardial hypertrophy . This was associated with mitigating the hypertrophy-associated gene signature , including reversing the myosin heavy chain isoform switch but in a pattern distinguishable from that associated with neovascularization-induced reversal . CONCLUSIONS : A system was developed suitable for identifying novel remodeling-reversing drugs operating in different pathways and for gaining insights into their mechanisms of action , exemplified here by uncoupling their vascular affects . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood-lymphocytes . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood lymphocytes was investigated in vitro . The effects of the antibiotics on the mitotic response of purified lymphocytes stimulated with P01584 varied according to the individual from whom the lymphocytes were obtained . At concentrations above those reported to be present in serum during conventional therapy ( 2-8 mg/l ) , there was a tendency for both minocycline and tetracycline to suppress the mitotic response . DB01017 was superior to tetracycline in this respect . However , at physiological concentrations the antibiotics either had no significant effect , suppressed the mitotic response ( minocycline at 2 mg/l with one of six donors ) , or enhanced the mitotic response ( tetracycline at 2 and 8 mg/l with four of six donors ) . The stimulatory effect of tetracycline was not demonstrated when lymphocytes were cultured in whole blood for up to seven days with the antibiotic alone . Similar effects of the antibiotics were observed when mononuclear cell fractions isolated from six donors were stimulated with an optimal concentration of phytohaemagglutinin ( PHA ) . Stimulation of lymphocytes in whole blood cultures with PHA in the presence of minocycline and tetracycline revealed that , under these culture conditions , the antibiotics could suppress the mitotic response of lymphocytes at physiological doses with cells from a majority of donors . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Delayed minocycline inhibits ischemia-activated matrix metalloproteinases 2 and 9 after experimental stroke . BACKGROUND : Matrix metalloproteinases 2 and 9 ( P08253 and P14780 ) are increased in the brain after experimental ischemic stroke in rats . These two proteases are involved with the degradation of the basal lamina and loss of stability of the blood brain barrier that occurs after ischemia and that is associated with thrombolytic therapy in ischemic stroke . DB01017 is a lipophilic tetracycline and is neuroprotective in several models of brain injury . DB01017 inhibits inflammation , apoptosis and extracellular matrix degradation . In this study we investigated whether delayed minocycline inhibits brain MMPs activated by ischemia in a model of temporary occlusion in Wistar rats . RESULTS : Both P08253 and P14780 were elevated in the ischemic tissue as compared to the contra-lateral hemisphere after 3 hours occlusion and 21 hours survival ( p < 0.0001 for P14780 ) . Intraperitoneal minocycline at 45 mg/kg concentration twice a day ( first dose immediately after the onset of reperfusion ) significantly reduced gelatinolytic activity of ischemia-elevated P08253 and P14780 ( p < 0.0003 ) . Treatment also reduced protein concentration of both enzymes ( p < 0.038 for P14780 and p < 0.018 for P08253 ) . In vitro incubation of minocycline in concentrations as low as 0.1 mug/ml with recombinant P08253 and P14780 impaired enzymatic activity and P14780 was more sensitive at lower minocycline concentrations ( p < 0.05 ) . CONCLUSION : DB01017 inhibits enzymatic activity of gelatin proteases activated by ischemia after experimental stroke and is likely to be selective for P14780 at low doses . DB01017 is a potential new therapeutic agent to acute treatment of ischemic stroke . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . DB01017 modulates cytokine and gene expression profiles in the brain after whole-body exposure to radiation . An effective countermeasure against radiation damage to normal tissues is urgently needed . The major goal of the present study was to determine if minocycline could modify the immunomodulatory effects of radiation on the brain . C57BL/6 mice were treated with minocycline intraperitoneally for 5 days beginning immediately before total-body exposure to 0 , 1 , 2 and 3 Gray ( Gy ) (60)Co γ-rays . Brains were collected on days 4 and 32 post-irradiation for cytokine and gene analyses . DB01017 treatment significantly increased the levels of interleukin ( IL ) -10 , P40933 and vascular endothelial growth factor ( P15692 ) in the brain on day 4 in one or more irradiated groups compared to radiation-alone ( p < 0.05 ) . P22301 is anti-inflammatory , P40933 can prevent apoptosis and P15692 is nuroprotective . On day 32 , the drug decreased IL-1β in the 2- Gy group ( p < 0.05 vs. 2-Gy alone ) ; this cytokine is implicated in immune-related central nervous system pathologies . Microarray analysis of brains on day 32 showed that while radiation increased expression of inflammatory genes such as Il1f10 , Il17 , Tnfrsf11b , Tnfsf12 , Il12b and Il1f8 , these were no longer up-regulated in the minocycline-treated groups . Similarly , the pro-apoptotic gene Bik and nitric oxide synthase producer ( Q8IVI9 ) were no longer up-regulated in the drug-treated groups . Pathway analysis based on gene data suggested that catenin-β1 and tumor suppressor-related transcription regulation were significantly activated by radiation and/or minocycline ( activation z-score > 2.0 ) . Overall , the data warrant further testing of minocycline as a potential neuroprotectant against radiation-induced damage . Effects of minocycline on cocaine sensitization and phosphorylation of GluR1 receptors in P09917 deficient mice . In wild-type ( WT ) mice , the antibiotic minocycline inhibits development of cocaine-induced locomotor sensitization . Some of the actions of minocycline may involve the P09917 ( 5- P28300 ) pathway . We used the model of 5- P28300 -deficient mice to investigate whether 5- P28300 participates in minocycline 's influence on the effects of cocaine . Locomotor sensitization was induced by 4 daily cocaine injections and the phosphorylation status of GluR1 glutamate receptors was assayed in brain samples . DB01017 failed to affect cocaine sensitization in 5- P28300 -deficient mice . In these mice , neither cocaine nor minocycline 4-day treatment altered GluR1 phosphorylation . In WT mice in which minocycline inhibited development of cocaine sensitization , a 4-day cocaine treatment increased GluR1 phosphorylation at both Ser831 and Ser845 sites in the frontal cortex but not the striatum ; further , this effect was prevented by minocycline . Under basal conditions and in response to a single cocaine injection the levels of GluR1 , GluR2 , and GluR3 AMPA receptor subunits did not differ between WT and 5- P28300 -deficient mice , but the response of GluR1 phosphorylation to a single cocaine injection was greater under the 5- P28300 deficiency . Hence , in WT mice GluR1 phosphorylation increased only in the frontal cortex and only at the Ser831 site . In 5- P28300 -deficient mice , acute cocaine injection increased both Ser831 and Ser845 phosphorylation both in the frontal cortex and in the striatum . We suggest that in studying minocycline 's action on cocaine 's effects and/or addiction in humans , it would be important to consider the characterization of the subjects ' 5- P28300 system . This article is part of a Special Issue entitled ' Trends in neuropharmacology : in memory of Erminio Costa ' . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . DB01017 attenuates hypoxia-inducible factor-1α expression correlated with modulation of p53 and AKT/ P42345 /p70S6K/ Q13541 pathway in ovarian cancer : in vitro and in vivo studies . Hypoxia-inducible factor ( HIF ) -1α is the key cellular survival protein under hypoxia , and is associated with tumor progression and angiogenesis . We have recently shown the inhibitory effects of minocycline on ovarian tumor growth correlated with attenuation of vascular endothelial growth factor ( P15692 ) and herein report a companion laboratory study to test if these effects were the result of HIF-1α inhibition . In vitro , human ovarian carcinoma cell lines ( A2780 , OVCAR-3 and SKOV-3 ) were utilized to examine the effect of minocycline on Q9BYW2 and its upstream pathway components to elucidate the underlying mechanism of action of minocycline . Mice harboring OVCAR-3 xenografts were treated with minocycline to assess the in vivo efficacy of minocycline in the context of Q9BYW2 . DB01017 negatively regulated HIF-1α protein levels in a concentration-dependent manner and induced its degradation by a mechanism that is independent of prolyl-hydroxylation . The inhibition of HIF-1α was found to be associated with up-regulation of endogenous p53 , a tumor suppressor with confirmed role in HIF-1α degradation . Further studies demonstrated that the effect of minocycline was not restricted to proteasomal degradation and that it also caused down-regulation of HIF-1α translation by suppressing the AKT/ P42345 /p70S6K/ Q13541 signaling pathway . DB01017 treatment of mice bearing established ovarian tumors , led to suppression of HIF-1α accompanied by up-regulation of p53 protein levels and inactivation of AKT/ P42345 /p70S6K/ Q13541 pathway . These data reveal the therapeutic potential of minocycline in ovarian cancer as an agent that targets the pro-oncogenic factor HIF-1α through multiple mechanisms . Overexpression of Q9BYF1 produces antitumor effects via inhibition of angiogenesis and tumor cell invasion in vivo and in vitro . Angiotensin II ( AngII ) is a multifunctional bioactive peptide in the renin-angiotensin system ( DB01367 ) . Q9BYF1 ( Q9BYF1 ) is a newly identified component of DB01367 . The role of AngII and Q9BYF1 in the metastasis of non-small cell lung cancer ( NSCLC ) and the effects on matrix metalloproteinases ( MMPs ) are still unknown . In the present study , the anti-invasive effect and mechanism of Q9BYF1 were investigated in vitro and in vivo . Results of a transwell assay showed that the overexpression of Q9BYF1 reduces the invasive ability of A549 cells in vitro . According to the results of qRT-PCR and western blot analysis , the inhibitory role of Q9BYF1 was mediated through the down-regulation of P08253 and P14780 . Additionally , we confirmed that the overexpression of Q9BYF1 inhibited cell growth and VEGFa production while simultaneously suppressing P12821 and angiotensin II type 1 receptor ( AT1R ) expression in human lung cancer xenografts . These results suggest that the overexpression of Q9BYF1 may potentially suppress the invasion and angiogenesis of NSCLC .

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Circulating hepatocyte growth factor as an independent prognostic factor of disseminated intravascular coagulation . BACKGROUND : P14210 ( P14210 ) , a pleiotropic factor regulating development and wound healing , is secreted as inactive pro- P14210 and is converted into active P14210 by coagulation serine proteases . P08581 overexpression can cause massive venous thrombi , and factor Xa is reported to release soluble P14210 from granulocytes . We hypothesized that a hypercoagulable condition , such as disseminated intravascular coagulation ( DIC ) , may increase circulating P14210 through active cleavage by coagulation serine proteases . METHODS : In 172 DIC-suspected patients , plasma levels of total and active P14210 , thrombin-antithrombin complex ( TAT ) , plasmin-antiplasmin complex ( PAP ) , and interleukin ( IL ) -6 were measured by ELISA . Active P14210 release in granulocytes was examined in patients with and without overt-DIC . P14210 -induced tissue factor expression in peripheral monocytes was measured by flow cytometry . RESULTS : Circulating levels of total and active P14210 correlated well with coagulopathy severity , including DIC score , D-dimer , TAT and PAP levels . P14210 positively correlated with P05231 and absolute neutrophil count . In contrast to the cancer group , P14210 levels were significantly increased in accordance with increased DIC scores in non-cancer group . Elevated circulating P14210 was an independent prognostic marker in the non-cancer group , while P14210 level failed to predict mortality in the cancer group . Amounts of P14210 released from stimulated granulocytes were not significantly different between overt-DIC and no overt-DIC patients . P14210 potentiated endotoxin-induced tissue factor expression of monocytes in vitro . CONCLUSION : These findings suggest that circulating P14210 is a potential laboratory marker reflecting coagulation activity and DIC prognosis in non-cancer patients and that P14210 may play a role in a vicious cycle of hypercoagulability . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . DB00175 -induced proangiogenic effects depend upon extracellular P09038 . The P04035 inhibitors ( statins ) have been shown to exert several protective effects on the vasculature that are unrelated to changes in the cholesterol profile , and to induce angiogenesis . The proangiogenic effect exerted by statins has been attributed to the activation of the PI3K/Akt pathway in endothelial cells ; however , it is unclear how statins activate this pathway . DB00175 -mediated activation of Akt and MAPK occurs rapidly ( within 10 min. ) and at low doses ( 10 nM ) . Here , we hypothesized that P09038 contributes to the proangiogenic effect of statins . We found that pravastatin , a hydrophilic statin , induced phosphorylation of the FGF receptor ( FGFR ) in human umbilical vein endothelial cells . SU5402 , an inhibitor of FGFR , abolished pravastatin-induced PI3K/Akt and MAPK activity . Likewise , anti- P09038 function-blocking antibodies inhibited Akt and MAPK activity . Moreover , depletion of extracellular P09038 by heparin prevented pravastatin-induced phosphorylation of Akt and MAPK . Treatment with P09038 antibody inhibited pravastatin-enhanced endothelial cell proliferation , migration and tube formation . These observations indicate that pravastatin exerts proangiogenic effects in endothelial cells depending upon the extracellular P09038 . P14210 plays a key role in insulin resistance-associated compensatory mechanisms . P01308 resistance is present in obesity and in type 2 diabetes and is associated with islet cell hyperplasia and hyperinsulinemia , but the driving forces behind this compensatory mechanism are incompletely understood . Previous data have suggested the involvement of an unknown circulating insulin resistance-related β-cell growth factor . In this context , looking for candidates to be a circulating factor , we realized that hepatocyte growth factor ( P14210 ) is a strong candidate as a link between insulin resistance and increased mass of islets/hyperinsulinemia . Our approach aimed to show a possible cause-effect relationship between increase in circulating P14210 levels and compensatory islet hyperplasia/hyperinsulinemia by showing the strength of the association , whether or not is a dose-dependent response , the temporality , consistency , plausibility , and reversibility of the association . In this regard , our data showed : 1 ) a strong and consistent correlation between P14210 and the compensatory mechanism in three animal models of insulin resistance ; 2 ) P14210 increases β-cell mass in a dose-dependent manner ; 3 ) blocking P14210 shuts down the compensatory mechanisms ; and 4 ) an increase in P14210 levels seems to precede the compensatory response associated with insulin resistance , indicating that these events occur in a sequential mode . Additionally , blockages of P08581 ( DB00134 ) worsen the impaired insulin-induced insulin signaling in liver of diet-induced obesity rats . Overall , our data indicate that P14210 is a growth factor playing a key role in islet mass increase and hyperinsulinemia in diet-induced obesity rats and suggest that the P14210 - DB00134 axis may have a role on insulin signaling in the liver . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . DB08865 for the treatment of patients with advanced non-small cell lung cancer . DB08865 is a potent small-molecule inhibitor of Q9UM73 ( anaplastic lymphoma kinase ; Q9UM73 ) and hepatocyte growth factor receptor ( P08581 , proto-oncogene c- DB00134 ) . A range of tumors , including subsets of non-small cell lung cancer ( NSCLC ) , anaplastic large cell lymphoma and inflammatory myofibroblastic tumors harbor an Q9UM73 rearrangement that leads to oncogenic activation of Q9UM73 . DB08865 has demonstrated preclinical and clinical activity against such malignancies through inhibition of Q9UM73 , and patients harboring Q9UM73 - rearranged NSCLC have demonstrated high response rates and prolonged progression-free survival in phase I and II studies . In August 2011 , crizotinib was approved for the treatment of advanced Q9UM73 -positive NSCLC . Gab1 but not Grb2 mediates tumor progression in DB00134 overexpressing colorectal cancer cells . P08581 ( DB00134 ) plays an important role in the progression of multiple cancer types . The overexpression of DB00134 in DLD-1 colon carcinoma cells with kirsten rat sarcoma oncogene homolog ( P01116 ) oncogene activation resulted in enhanced subcutaneous and orthotopic tumor growth rate and increased metastatic potential . To elucidate the mechanism of this effect , we stably expressed kinase-inactive DB00134 (K1110A) , Src homology 2 ( SH2 ) -binding domain-inactive DB00134 (Y1349/1356F) , growth factor receptor-bound protein 2 ( Grb2 ) non-binding DB00134 (N1358H) and mutant receptors with ability to selectively recruit signaling proteins Grb2 , src homology domain c-terminal adaptor homolog ( Shc ) , phospholipase c-gamma ( PLCgamma ) and p85 phosphatidyl inositol 3 kinase . As subcutaneous implants , DLD-1 cells that expressed the majority of these receptor constructs failed to recapitulate the tumor growth-enhancing effect of the wild-type DB00134 receptor . The Grb2- and Shc-recruiting DB00134 mutants demonstrated slight but consistent tumor-suppressive activity , whereas the expression of N1358H mutant stimulated tumor growth rate comparable with the wild-type receptor . This suggests that direct Grb2/Shc binding does not contribute to the tumor progression activity of DB00134 receptor . The tumors expressing Grb2- and Shc-recruiting DB00134 receptors demonstrated a marked loss in Grb2-associated adaptor protein 1 ( Gab1 ) protein levels , which was not observed in the cell lines , consistent with a post-translationally regulated process . Moreover , a moderate level of Gab1 overexpression stimulated tumor growth . The findings suggest a delicate balance for intact Y1349/1356 SH2-binding domain to mediate the tumor progression activity of the coactivated DB00134 -rat sarcoma oncogene homolog ( DB01367 ) pathways . Selectivity for specific adaptor protein involvement may be the key that determines the tissue- and cell-type specificity of DB00134 -mediated tumorigenicity in human cancers . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . HSulf-1 modulates P14210 -mediated tumor cell invasion and signaling in head and neck squamous carcinoma . Recently , we cloned a novel sulfatase domain-containing downregulated gene , HSulf-1 , which modulates heparin-binding growth factor signaling in ovarian cancer . Based on the pilot data showing the loss of HSulf-1 in head and neck squamous cell carcinoma cell lines ( SCCHN ) , we sought to employ SCCHN as a model to define the role of HSulf-1 in the molecular regulation of tumorigenicity . Three SCCHN lines ( 012SCC , WMMSCC , and 015SCC ) had no detectable HSulf-1 mRNA . Clonal lines of HSulf-1-expressing 012SCC attenuated the activation of P29323 /mitogen-activated protein kinase ( MAPK ) signaling mediated by fibroblast growth factor ( P09038 ) and both P29323 /MAPK and Akt signaling mediated by hepatocyte growth factor ( P14210 ) . Consistent with this downregulation , phosphorylation of P08581 , c- DB00134 , which is frequently overexpressed in SCCHN , was also attenuated in HSulf-1 clonal 012SCC cell lines . P14210 markedly enhanced the motility and migration of vector-transfected cells in a transwell invasion chamber . However , P14210 -mediated motility and invasion was attenuated in HSulf-1 clonal 012SCC cell lines . In addition , transfected cells displayed significant growth inhibition concomitant with a decrease in mitogenicity , as measured by thymidine incorporation and increased sensitivity to staurosporine- and cisplatin-induced apoptosis . These data suggest that HSulf-1 normally functions as a negative regulator in cell growth and loss of HSulf-1 in SCCHN potentiates growth factor signaling , enhances motility , invasiveness and inhibits stress-induced apoptosis , with a resulting increase in tumorigenicity . Differentiation and proliferation of primary rat hepatocytes cultured as spheroids . We studied spheroid ( multicellular aggregate ) formation by hepatocytes and the expression of liver-specific functions such as albumin secretion when hepatocytes were cultured with various extracellular matrices . Hepatocytes cultured on Primaria(R) and poly-D-lysine coated dishes , and in the presence of a polymer , Eudragit , formed spheroids , and they also exhibited higher liver-specific functions and poor growth compared to monolayer cultures . The results indicated that the cell morphological change and cell-cell interaction caused by the spheroid formation were key factors promoting the expression of the liver-specific functions . To elucidate the mechanism underlying the poor growth in spheroids , we examined the P14210 signaling pathway . Phosphorylation and down-regulation of the P08581 ( c- DB00134 proto-oncogene product ) were observed for the cells from both monolayer and spheroid cultures , but Ras activation was partly blocked in spheroids . Furthermore , we found that CDK inhibitors , P38936 and p27 , were highly expressed in spheroids . These results suggested that the reduced Ras signaling and high expression of the CDK inhibitors might cause the lower growth in spheroids . We then examined the relationship between liver-enriched transcription factors ( C/EBPalpha and beta ) and liver-specific functions . The results revealed that the high expression of C/EBPalpha was maintained during cultures when hepatocytes formed spheroids . Antisense oligonucleotides of C/EBPalpha repressed albumin secretion and the expression of P38936 , suggesting that the transcription factor , C/EBPalpha , may play a crucial role in the growth and differentiation of hepatocytes in spheroids . [ c- DB00134 signaling pathway participating in the gefitinib resistance of different gene types of non-small cell lung cancer cells induced by P14210 in vitro ] . BACKGROUND AND OBJECTIVE : It has been known that hepatocyte growth factor ( P14210 ) induces gefitinib resistance in non-small cell lung cancer ( NSCLC ) cells . The possible mechanism may be related to the activation of the P08581 c- DB00134 . The aim of this study is to investigate the involvement of c- DB00134 and its downstream signaling pathway in the P14210 -induced gefitinib resistance of NSCLC cells with different epidermal growth factor receptor ( P00533 ) gene types . METHODS : NSCLC cell lines with different P00533 genes ( PC-9 , Q8NBP7 /R , H292 , and A549 ) were selected and induced by P14210 . Cell survival was determined by MTT assay and the expression of DB00134 and downstream signaling proteins were examined by Western blot . RESULTS : Gefitinib inhibited the cell growth of Q8NBP7 , H292 , and A549 cell lines in a dose-dependent manner . The concentration-survival curve notably shifted to the right when induced by P14210 . The apoptotic rate was lower when the cells were treated with P14210 and gefitinib than when these cells were treated with gefitinib alone ( P < 0.05 ) , particularly in Q8NBP7 , H292 , and A549 cells , but not in Q8NBP7 /R . P14210 stimulated the phosphorylation of DB00134 and downstream signaling proteins in Q8NBP7 , H292 , Q8NBP7 /R , and A549 cell lines . p- DB00134 , p-Akt , p-Stat3 , and p-Erk1/2 expressions were higher when the cells were treated with P14210 and gefitinib than when these cells were treated with gefitinib alone , particularly in Q8NBP7 , H292 , and A549 cells , but not in Q8NBP7 /R . CONCLUSIONS : c- DB00134 and its downstream signaling pathway possibly participated in the P14210 -induced gefitinib resistance in NSCLC cells with different P00533 gene types . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Molecular mechanisms of cell proliferation induced by low power laser irradiation . Low power laser irradiation ( LPLI ) promotes proliferation of multiple cells , which ( especially red and near infrared light ) is mainly through the activation of mitochondrial respiratory chain and the initiation of cellular signaling . Recently , the signaling proteins involved in LPLI-induced proliferation merit special attention , some of which are regulated by mitochondrial signaling . P08581 ( c- DB00134 ) , a member of tyrosine protein kinase receptors ( TPKR ) , is phosphorylated during LPLI-induced proliferation , but tumor necrosis factor alpha ( P01375 ) receptor has not been affected . Activated TPKR could activate its downstream signaling elements , like Ras/Raf/MEK/ P29323 , PI3K/Akt/ P06730 , PI3K/Akt/ P29474 and P98160 -gamma/PKC pathways . Other two pathways , DeltaPsim/ DB00171 / DB02527 /JNK/AP-1 and ROS/Src , are also involved in LPLI-induced proliferation . LPLI-induced cell cycle progression can be regulated by the activation or elevated expressions of cell cycle-specific proteins . Furthermore , LPLI induces the synthesis or release of many molecules , like growth factors , interleukins , inflammatory cytokines and others , which are related to promotive effects of LPLI . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . P08581 tyrosine kinase met is a substrate of the receptor protein-tyrosine phosphatase Q12913 . The receptor protein-tyrosine phosphatase ( PTP ) Q12913 ( CD148/PTP-eta ) has been implicated in the regulation of cell growth , differentiation , and transformation , and most recently has been identified as a potential tumor suppressor gene mutated in colon , lung , and breast cancers . We have generated constructs comprising the cytoplasmic segment of Q12913 fused to the maltose-binding protein to identify potential substrates and thereby suggest a physiological function for Q12913 . We have shown that the substrate-trapping mutant form of Q12913 interacted with a small subset of tyrosine-phosphorylated proteins from lysates of the human breast tumor cell lines MDA-MB-231 , T-47D , and T-47D/ DB00134 and have identified the hepatocyte growth factor/scatter factor receptor DB00134 , the adapter protein Gab1 , and the junctional component O60716 as potential substrates . Following ligand stimulation , phosphorylation of specific tyrosyl residues in DB00134 induces mitogenic , motogenic , and morphogenic responses . When co-expressed in 293 cells , the full-length substrate-trapping mutant form of Q12913 formed a stable complex with the chimeric receptor colony stimulating factor 1 ( P04141 ) - DB00134 and wild type Q12913 dephosphorylated P04141 - DB00134 . Furthermore , we observed that Q12913 preferentially dephosphorylated a Gab1 binding site ( DB00135 (1349) ) and a COOH-terminal tyrosine implicated in morphogenesis ( DB00135 (1365) ) , whereas tyrosine residues in the activation loop of DB00134 ( DB00135 (1230) , DB00135 (1234) , and DB00135 (1235) ) were not preferred targets of the PTP . The ability of Q12913 preferentially to dephosphorylate particular tyrosine residues that are required for DB00134 -induced signaling suggests that Q12913 may function in controlling the specificity of signals induced by this PTK , rather than as a simple " off-switch " to counteract PTK activity . P14210 and P04626 /neu downregulate expression of apoptosis-inducing factor in non-small cell lung cancer . Our previous study showed that patients with advanced stages of non-small cell lung cancer ( NSCLC ) were frequently detected with upregulation of hepatocyte growth factor ( P14210 ) . In vitro , P14210 reduced expression of apoptosis-inducing factor ( O95831 ) and cisplatin sensitivity in NSCLC cells . The effect of P14210 was via P08581 ( c-MET ) and the downstream effector , focal adhesion kinase ( Q05397 ) . In this study , we determined the prognostic value of O95831 in NSCLC patients . O95831 expression was determined by immunohistochemistry and immunoblotting . Our data show that O95831 expression was associated with better prognosis . Expression of O95831 inversely correlated with that of positive NSCLC markers , e.g. , dihydrodiol dehydrogenase ( Q04828 ) , c-MET , short oncostatin M receptor ( OSMRs ) , matrix metalloproteinase ( MMP ) -1 , and P04626 /neu , which were closely associated with drug resistance , tumor recurrence , metastasis and poor prognosis . Noteworthy , silence of P04626 /neu gene expression increases O95831 level and drug sensitivity . Addition of P14210 inhibits O95831 expression in P04626 /neu-silenced cells . These results suggested that both P14210 and P04626 /neu affect drug resistance by regulating O95831 expression in NSCLC .

MH_train_1017

MH_train_1017

MH_train_1017

interacts_with DB00163?

multiple_choice

Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . Early vitamin E supplementation attenuates diabetes-associated vascular dysfunction and the rise in protein kinase C-beta in mesenteric artery and ameliorates wall stiffness in femoral artery of Wistar rats . AIMS/HYPOTHESIS : The impact of early vitamin E supplementation on vascular function in diabetes remains unresolved . Therefore , we examined the effects of vitamin E on functional and structural parameters and on chemical markers that are disturbed in diabetes in mesenteric and femoral arteries . METHODS : Segments of both arteries , taken from control and 8-week-old streptozotocin diabetic Wistar rats that were treated or not with vitamin E , were mounted on wire and pressure myographs , after which endothelium-dependent and -independent vasodilation was assessed . Passive mechanical wall properties and the localisation and levels of protein kinase C ( PKC ) -beta(2) and P51606 were evaluated in these vessels . RESULTS : DB00163 supplementation was associated with improved endothelium-dependent and -independent vasodilatation in mesenteric arteries from diabetic rats . Impaired endothelium-dependent vasodilatation in diabetic mesenteric vessels was associated with P05771 (2) up-regulation and this was prevented by vitamin E supplementation . Increased P51606 accumulation and plasma isoprostane levels in diabetic rats were not changed by vitamin E . In the femoral artery , vitamin E supplementation had no effect on endothelium-dependent or -independent vasodilatation , but did prevent the wall stiffening associated with diabetes . CONCLUSIONS/INTERPRETATION : Early vitamin E supplementation has a beneficial effect on diabetes-induced endothelial dysfunction in resistance arteries . This benefit may arise from a direct effect on smooth muscle function , as a result of inhibition of the P05771 (2) isoform by vitamin E . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Flavonoids inhibit the oxidative modification of low density lipoproteins by macrophages . Low density lipoproteins ( LDL ) can be oxidatively modified in vitro by macrophages and certain other cell types so that macrophages will take them up much faster . This process may be important in the formation of cholesterol-laden foam cells derived from macrophages in atherosclerotic lesions . In this study , we have shown that certain flavonoids , plant constituents found in the diet , are potent inhibitors of the modification of 125I-labelled LDL by macrophages , with IC50 values in the micromolar range ( e.g. morin and fisetin 1 microM ; quercetin and gossypetin 2 microM ) . The potencies of individual flavonoids in inhibiting LDL modification did not correlate with their previously determined potencies as inhibitors of P09917 and cyclo-oxygenase . The modification of LDL by macrophages exhibits a lag period of about 4-6 hr before enhanced uptake is detected . During this time , there is a rapid depletion in its content of DB00163 ( an endogenous antioxidant found in lipoproteins ) followed by a large increase in the level of hydroperoxides . The flavonoids conserved the DB00163 content of LDL and delayed the onset of detectable lipid peroxidation . Flavonoids also inhibited the cell-free oxidation of LDL mediated by CuSO4 . These findings raise the possibility that flavonoids may protect LDL against oxidation in atherosclerotic lesions and may therefore be natural anti-atherosclerotic components of the diet , although this will depend to a large extent on their pharmacokinetics . Tocotrienols activate the steroid and xenobiotic receptor , O75469 , and selectively regulate expression of its target genes . DB00163 is an essential nutrient with antioxidant activity . DB00163 is comprised of eight members , alpha- , beta- , gamma- , and delta-tocopherols and alpha- , beta- , gamma- , and delta-tocotrienols . All forms of vitamin E are initially metabolized by omega-oxidation , which is catalyzed by cytochrome P450 enzymes . The steroid and xenobiotic receptor ( O75469 ) is a nuclear receptor that regulates drug clearance in the liver and intestine via induction of genes involved in drug and xenobiotic metabolism . We show here that all four tocotrienols specifically bind to and activate O75469 , whereas tocopherols neither bind nor activate . Surprisingly , tocotrienols show tissue-specific induction of O75469 target genes , particularly P08684 . Tocotrienols up-regulate expression of P08684 but not P22309 ( P22309 ) or multidrug resistance protein-1 ( P08183 ) in primary hepatocytes . In contrast , tocotrienols induce P08183 and P22309 but not P08684 expression in intestinal LS180 cells . We found that nuclear receptor corepressor ( NCoR ) is expressed at relatively high levels in intestinal LS180 cells compared with primary hepatocytes . The unliganded O75469 interacts with NCoR , and this interaction is only partially disrupted by tocotrienols . Expression of a dominant-negative NCoR enhanced the ability of tocotrienols to induce P08684 in LS180 cells , suggesting that NCoR plays an important role in tissue-specific gene regulation by O75469 . Our findings provide a molecular mechanism explaining how vitamin supplements affect the absorption and effectiveness of drugs . Knowledge of drug-nutrient interactions may help reduce the incidence of decreased drug efficacy . Inhibitory effects of a phosphate diester of DB00163 and ascorbic acid ( EPC- P04264 ) on myocardial infarction in rats . The inhibitory effect of a phosphate diester of DB00163 and ascorbic acid ( EPC- P04264 ) was examined in myocardial infarction induced in rats , in comparison with a selective P09917 inhibitor , AA-861 . EPC- P04264 significantly reduced the infarct size at 24 and 48 h after ligation , whereas AA-861 reduced it only at 48 h after ligation . In in-vitro experiments , EPC- P04264 inhibited not only superoxide anion generation ( IC50 = 4.2 x 10(-5) M ) , but also acid phosphatase activity ( IC50 = 2.4 x 10(-5) M ) in rat polymorphonuclear leukocytes in a concentration-dependent manner , while AA-861 showed marginal effects on both actions . These results indicated that EPC- P04264 induced cardioprotective effects by affecting neutrophil functions by inhibition of generation of superoxide-anion generation and acid-phosphatase activity . The mechanism of the reduction of the infarct size by EPC- P04264 differed from that of AA-861 , which latter inhibited P09917 and the formation of leukotriene B4 . Anti-allergic effects of nilotinib on mast cell-mediated anaphylaxis like reactions . DB04868 is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of P11274 - P00519 -positive chronic myelogenous leukemia . However , its effect on mast cell-mediated anaphylactic reaction is still not known . The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action . DB04868 administration prevented systemic anaphylaxis in mice , mediated by compound 48/80 , in a dose- and time-dependent manner . Also , nilotinib significantly inhibited ( P < 0.05 ) allergic paw edema in rats . Furthermore , nilotinib significantly decreased ( P < 0.05 ) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner . In addition , nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin . Moreover , nilotinib attenuated the secretion of pro-inflammatory cytokine , tumor necrosis factor ( P01375 ) -α expression in the rat peritoneal mast cells . These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Construction , expression and characterization of tissue-type plasminogen activator mutants . Three tissue-type plasminogen activator ( t-PA ) mutants were constructed by recombinant and site-directed mutagenesis techniques . They are del(296-302) with deletion of P05121 binding site , N117Q/N184Q with deglycosylation of P04264 and K2 domains , and their combination mutant designated as GGI . Then these three mutants were successfully transiently expressed in COS-7 cells , and GGI was further stably expressed in CHO cells . The biological characterization of the expression products indicated that del(296-302) and GGI possessed the resistance to inhibition by P05121 . In addition , the specific activity of GGI was increased by about 46 % , the plasma half-life was prolonged by about one fold , while its affinity for fibrin was not affected . Screening of hub genes and pathways in colorectal cancer with microarray technology . Here we intend to identify key genes and pathways in the pathogenesis of colorectal cancer ( CRC ) through analyzing microarray data with bioinformatic tools . The gene expression profile dataset GSE23878 was downloaded from Gene Expression Omnibus and differentially expressed genes ( DEGs ) were screened out using Student 's t-test . GO function and KEGG pathway enrichment analyses were performed for these DEGs with the DAVID online tool . Interaction network was constructed among the over-represented pathways based on the protein-protein interactions within the pathways . Besides , the protein interaction information obtained from HPRD database were applied to constructed protein-protein interaction networks among the DEGs and hub genes and function module were screened out . A total of 2,296 DEGs were obtained and they were enriched in 34 pathways . An interaction network was constructed among 32 pathways , in which p53 signaling pathway acted as the hub pathway as it showed the highest node degree . The protein-protein interaction network comprised 1,481 interaction relationships among 332 genes which included 40 DEGs . Further analysis revealed that theses DEGs formed 7 function modules and many genes , such as P09619 , MET , Q14332 , P24385 , P05771 , Q15052 , P14923 , P09544 , P41221 and O96014 were key genes in the networks . The DEGs and disturbed biological functions uncovered in present study may play important roles in the development of CRC and can contribute to the understanding on molecular mechanisms of CRC . Further these DEGs we obtained can be acted as potential biomarkers for diagnosis and therapy of CRC . Determination of ancestral allele for possible human cancer-associated polymorphisms . To determine ancestral allele in possible cancer-associated polymorphisms , DNA samples from 10 chimpanzees ( Pan troglodytes ) were sequenced for alleles corresponding to 17 polymorphisms : 8 short tandem repeats [ P18510 ( alias IL-1RA ) variable number tandem repeat ( VNTR ) ; P04818 ( previously TS ) VNTR ; AR CAG repeat ; dinucleotide repeats of P22309 , IGF1 , P01579 ( alias P01579 ) , P03372 ( alias P03372 ) , and P00533 ] and 9 single nucleotide polymorphisms ( P03956 -1607 1G/2G , P08254 -1171 5A/6A , O15527 Ser326Cys , P05091 Gly487Lys , P04637 Arg72Pro , Q9UNQ0 Gln141Lys , P16455 Leu84Phe , P04179 Ala-9Val , and P42898 Ala222Val ) . No chimpanzee polymorphism corresponded to human P18510 VNTR ; the ancestral allele was a repeat lost in humans . Dinucleotide repeat polymorphisms of IGF1 , P01579 , P03372 , and P00533 were shared by chimpanzees , but the length of repeats tended to be longer in humans than in chimpanzees . This tendency was particularly evident for IGF1 . All of the SNPs tested are human-specific nucleotide changes . The ancestral allele 7A was shown to be lost in P08254 -1171 5A/6A . Thus , all of the possible cancer-associated polymorphisms tested have human-specific alleles , and the ancestral allele is lost in three polymorphisms ( P18510 VNTR , P22309 CA repeat , and P08254 -1171 5A/6A ) , suggesting a possible involvement of human-specific alleles in cancer susceptibility . The 80th anniversary of vitamin E : beyond its antioxidant properties . Molecules provided with an antioxidant function may have additional properties , the latter being sometimes of greater importance than the former . In the last ten years , DB00163 has revealed precise cellular functions , some of which are independent of its antioxidant/radical scavenging ability . At the posttranslational level , DB00163 inhibits protein kinase C and P09917 and activates protein phosphatase 2A and diacylglycerol kinase . Some genes ( P16671 , alpha-TTP , alpha-tropomyosin , and collagenase ) are affected by DB00163 at the transcriptional level . alpha-Tocopherol also induces inhibition of cell proliferation , platelet aggregation and monocyte adhesion . These effects are unrelated to the antioxidant activity of vitamin E , but rather are believed to be a result of specific interactions of vitamin E with components of the cell , e. g. proteins , enzymes and membranes . This review focuses on novel non-antioxidant functions of DB00163 and discusses the possibility that many of the effects previously attributed to the antioxidant functions can also be explained by non-antioxidant mechanisms . DB00163 , P09917 and oxidative stress in haemodialysis patients : facts , not fancies . Gene expression profiles of B-lineage adult acute lymphocytic leukemia reveal genetic patterns that identify lineage derivation and distinct mechanisms of transformation . PURPOSE : To characterize gene expression signatures in acute lymphocytic leukemia ( ALL ) cells associated with known genotypic abnormalities in adult patients . EXPERIMENTAL DESIGN : Gene expression profiles from 128 adult patients with newly diagnosed ALL were characterized using high-density oligonucleotide microarrays . All patients were enrolled in the Italian GIMEMA multicenter clinical trial 0496 and samples had > 90 % leukemic cells . Uniform phenotypic , cytogenetic , and molecular data were also available for all cases . RESULTS : T-lineage ALL was characterized by a homogeneous gene expression pattern , whereas several subgroups of B-lineage ALL were evident . Within B-lineage ALL , distinct signatures were associated with Q03164 / P51825 and P15923 / P40424 gene rearrangements . Expression profiles associated with Q03164 / P51825 and P15923 / P40424 are similar in adults and children . P11274 / P00519 + gene expression pattern was more heterogeneous and was most similar to ALL without known molecular rearrangements . We also identified a set of 83 genes that were highly expressed in leukemia blasts from patients without known molecular abnormalities who subsequently relapsed following therapy . Supervised analysis of kinase genes revealed a high-level P36888 expression in a subset of cases without molecular rearrangements . Two other kinases ( P05771 and Q08345 ) were highly expressed in cases without molecular rearrangements , as well as in P11274 / P00519 -positive ALL . CONCLUSIONS : Genomic signatures are associated with phenotypically and molecularly well defined subgroups of adult ALL . Genomic profiling also identifies genes associated with poor outcome in cases without molecular aberrations and specific genes that may be new therapeutic targets in adult ALL . Lipoxygenase pathway mediates increases of airway resistance and lung inflation induced by exposure to nanotitanium dioxide in rats . Nanotitanium dioxide particle ( nTiO2 ) inhalation has been reported to induce lung parenchymal injury . After inhalation of nTiO2 , we monitored changes in P09917 , endothelial nitric oxide synthase ( P29474 ) , and inducible nitric oxide synthase ( P35228 ) mRNA in rat lung tissue . Lung function parameters include specific airway resistance ( SRaw ) , peak expiratory flow rate ( PEF ) , functional residual capacity ( FRC ) , and lung compliance ( Cchord ) ; blood white blood cell count ( WBC ) , nitric oxide ( NO ) , hydrogen peroxide , and lactic dehydrogenase ( LDH ) ; and lung lavage leukotriene C4 , interleukin 6 ( P05231 ) , tumor necrotic factor α ( TNFα ) , hydroxyl radicals , and NO . Leukotriene receptor antagonist MK571 and P09917 inhibitor MK886 were used for pharmacologic intervention . Compared to control , nTiO2 exposure induced near 5-fold increase in P09917 mRNA expression in lung tissue . P35228 mRNA increased while P29474 mRNA decreased . Lavage leukotriene C4 ; P05231 ; TNFα ; NO ; hydroxyl radicals ; and blood WBC , NO , hydrogen peroxide , and LDH levels rose . Obstructive ventilatory insufficiency was observed . MK571 and MK886 both attenuated the systemic inflammation and lung function changes . We conclude that inhaled nTiO2 induces systemic inflammation , cytokine release , and oxidative and nitrosative stress in the lung . The lipoxygenase pathway products , mediated by oxygen radicals and WBC , play a critical role in the obstructive ventilatory insufficiency induced by nTiO2 . DB09073 : first global approval . DB09073 ( Ibrance® ) is an oral , reversible , selective , small-molecule inhibitor of cyclin-dependent kinases ( CDK ) 4 and Q00534 developed by Pfizer for the treatment of cancer . CDKs are important modulators of cell cycle entry and progression in response to growth signals , and inhibition of these kinases with palbociclib could enhance the activity of other anticancer drugs in tolerable regimens . DB09073 , in combination with letrozole , was recently approved in the US for the first-line treatment of advanced breast cancer . Phase III development is underway worldwide investigating its use as first-line treatment in advanced breast cancer , as well as treatment of recurrent or advanced breast cancer and high-risk , early-stage breast cancer . A phase II trial is underway in the USA for non-small cell lung cancer under a US National Cancer Institute-funded research collaboration , and several phase I and II investigations are being conducted for various other solid tumour types and haematological malignancies . This article summarizes the milestones in the development of palbociclib leading to this first approval for use in postmenopausal women with estrogen-positive , human epidermal growth factor receptor ( HER ) 2-negative advanced breast cancer as initial endocrine-based therapy for their metastatic disease . Cannabinoid receptor activation induces apoptosis through tumor necrosis factor alpha-mediated ceramide de novo synthesis in colon cancer cells . PURPOSE : Cannabinoids have been recently proposed as a new family of potential antitumor agents . The present study was undertaken to investigate the expression of the two cannabinoid receptors , P21554 and CB2 , in colorectal cancer and to provide new insight into the molecular pathways underlying the apoptotic activity induced by their activation . EXPERIMENTAL DESIGN : Cannabinoid receptor expression was investigated in both human cancer specimens and in the DLD-1 and HT29 colon cancer cell lines . The effects of the P21554 agonist arachinodyl-2'-chloroethylamide and the CB2 agonist N-cyclopentyl-7-methyl-1-(2-morpholin-4-ylethyl)-1,8-naphthyridin-4(1H)-on-3-carboxamide ( CB13 ) on tumor cell apoptosis and ceramide and tumor necrosis factor ( P01375 ) -alpha production were evaluated . The knockdown of P01375 mRNA was obtained with the use of selective small interfering RNA . RESULTS : We show that the P21554 receptor was mainly expressed in human normal colonic epithelium whereas tumor tissue was strongly positive for the CB2 receptor . The activation of the P21554 and , more efficiently , of the CB2 receptors induced apoptosis and increased ceramide levels in the DLD-1 and HT29 cells . Apoptosis was prevented by the pharmacologic inhibition of ceramide de novo synthesis . The CB2 agonist CB13 also reduced the growth of DLD-1 cells in a mouse model of colon cancer . The knockdown of P01375 mRNA abrogated the ceramide increase and , therefore , the apoptotic effect induced by cannabinoid receptor activation . CONCLUSIONS : The present study shows that either P21554 or CB2 receptor activation induces apoptosis through ceramide de novo synthesis in colon cancer cells . Our data unveiled , for the first time , that P01375 acts as a link between cannabinoid receptor activation and ceramide production . Effect of the hemoregulatory peptide (pEEDCK)2 (pyroGlu- DB00142 - DB00128 - DB00151 -Lys)2 and MIP-1alpha is reduced in bone marrow cultures from patients with chronic myeloid leukemia ( CML ) . The granulocyte-derived hemoregulatory peptide pyroGlu- DB00142 - DB00128 - DB00151 -Lys = pEEDCK is known to keep hematopoietic cells quiescent . When oxidized to its dimeric form (pEEDCK)2 , it activates growth of hematopoietic progenitors in association with stroma-derived cytokines . (pEEDCK)2 has a DB00151 - DB00151 motif which is also a typical feature of the macrophage inflammatory protein ( MIP-1alpha ) . The present study was designed to analyze differences between the response of normal and leukemic progenitor cells to (pEEDCK)2 or MIP-1alpha . When long-term bone marrow cultures ( LTBMCs ) were incubated with (pEEDCK)2 or MIP-1alpha and/or cytokines , the stimulatory effect on colony-forming units-granulocyte/erythroid/macrophage/megakaryocyte of LTBMC from chronic myeloid leukemia ( CML ) patients was less than 50 % compared to LTBMC from healthy humans . No difference in oncogene expression could be observed in LTBMC from CML patients regarding reduction of Philadelphia chromosome-associated transcription of the P11274 - P00519 gene . With respect to the expression of growth and differentiation-associated genes ( Galpha16 , P09917 , phospholipaseA2 , c-kit , and P28906 ) , which were analyzed from LTBMC by semiquantitative reverse transcriptase-polymerase chain reaction , the same transcription rate was observed in CML patients and healthy donors . However , two isoforms of a key enzyme of oxidative metabolism , carnitine palmitoyltransferase ( P50416 and Q92523 ) , showed 50-fold higher expression rates in LTBMC cells of healthy donors compared to CML patients . It is known that a decrease in oxidative metabolism is associated with an increase in redox equivalents in malignancy . This might result in a reduction of disulphide bonds in (pEEDCK)2 or MIP-1alpha , thus inducing a downregulation of these factors in bone marrow from CML patients . P21554 and CB2 cannabinoid receptors differentially regulate the production of reactive oxygen species by macrophages . AIMS : We investigated the mechanism by which cannabinoid receptors-1 ( P21554 ) and -2 ( CB2 ) modulate inflammatory activities of macrophages . METHODS AND RESULTS : Real-time polymerase chain reaction showed the predominant CB2 expression in freshly isolated human monocytes . PMA , a potent inducer of differentiation , upregulated P21554 and increased P21554 :CB2 transcript ratio from 1:17.5 to 1:3 in 5 days of culture . Immunohistochemistry showed that P21554 protein was colocalized in P34810 - and P16671 -positive macrophages in human atheroma . Through selective expression of P21554 or CB2 to thioglycollate-elicited peritoneal macrophages , we proved that P21554 and CB2 mediate opposing influences on the production of reactive oxygen species ( ROS ) . Flow cytometry showed that cannabinoid-induced ROS production by macrophages was P21554 -dependent . Immunoblotting assays confirmed that macrophage P21554 , not CB2 , induced phosphorylation of p38-mitogen-activated protein kinase , which modulated ROS production and the subsequent synthesis of tumour necrosis factor-alpha and monocyte chemoattractant protein-1 . Pull-down assays showed that the Ras family small G protein , Rap1 was activated by CB2 . Dominant-negative Rap1 profoundly enhanced P21554 -dependent ROS production by macrophages , suggesting CB2 Rap1-dependently inhibits P21554 -stimulated ROS production . CONCLUSION : P21554 promotes pro-inflammatory responses of macrophages through ROS production , which is negatively regulated by CB2 through Rap1 activation . Blocking P21554 together with selective activation of CB2 may suppress pro-inflammatory responses of macrophages . Cytoprotective properties of DB00163 are related to gene regulation in cultured D-galactosamine-treated human hepatocytes . DB00163 ( DB00163 ) has demonstrated antioxidant activity and gene-regulatory properties . d-Galactosamine ( D-GalN ) -induced cell death is mediated by nitric oxide in hepatocytes , and it is associated with hepatic steatosis . The beneficial properties of DB00163 and their relation to oxidative stress and gene regulation were assessed in D-GalN-induced cell death . Hepatocytes were isolated from human liver resections by a collagenase perfusion technique . alpha-Tocopherol ( 50 microM ) was administered at the advanced stages ( 10 h ) of D-GalN-induced cell death in cultured hepatocytes . Cell death , oxidative stress , DB00163 metabolism , and NF-kappaB- , pregnane X receptor ( O75469 ) - , and peroxisome proliferator-activated receptor ( Q07869 ) -associated gene regulation were estimated in the hepatocytes . D-GalN increased cell death and DB00163 metabolism . alpha-Tocopherol exerted a moderate beneficial effect against apoptosis and necrosis induced by D-GalN . Induction ( rifampicin ) or inhibition ( ketoconazole ) of DB00163 metabolism and overexpression of O75469 showed that the increase in O75469 -related P08684 expression caused by DB00163 enhanced cell death in hepatocytes . Nevertheless , the reduction in NF-kappaB activation and inducible nitric oxide synthase expression and the enhancement of Q07869 and carnitine palmitoyl transferase gene expression by DB00163 may be relevant for cell survival . In conclusion , the cytoprotective properties of DB00163 are mostly related to gene regulation rather than to antioxidant activity in toxin-induced cell death in hepatocytes . 25-Hydroxycholesterol is not a ligand for the orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) . The orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) is involved in the transcriptional regulation of all the steroid hydroxylase genes , and also regulates the transcription of the genes for Müllerian Inhibitory substance ( P03971 ) , alpha subunit of glycoprotein hormone , LHbeta , oxytocin , P30968 , Q01718 , prolactin receptor , DAX-1 , and steroidogenic acute regulatory protein . Other members of the nuclear receptor gene family , including steroid hormone , thyroid hormone , retinoic acid , Q07869 , and vitamin D receptors must bind ligand to activate transcription , but Q13285 has been considered to be an orphan nuclear receptor because , when identified , it had no known ligand . A recent publication suggested that transcriptional regulation by Q13285 , expressed in a non-steroidogenic CV-1 cells , could be activated by oxysterols suggesting that these compounds could serve as natural ligands for Q13285 . We now demonstrate that 25-hydroxycholesterol , either added exogenously or synthesized endogenously in steroidogenic mouse Leydig MA-10 cells , did not act as a ligand for Q13285 , as it did not increase transcription from six different Q13285 -dependent DNA sequences . Furthermore , the abundance of these oxysterols in MA-10 cells was much less than concentrations needed for activation of Q13285 in CV-1 cells , indicating that Q13285 is not constitutively bound by ligand in MA-10 cells . Thus , in steroidogenic cells , transcriptional regulation of the steroid hydroxylase genes by Q13285 does not depend upon the presence of 25-hydroxycholesterol , and is not modified by its presence . Dedifferentiated chondrosarcoma mimicking a giant cell tumor . Is this low grade dedifferentiated chondrosarcoma ? We report a very rare case of a dedifferentiated chondrosarcoma mimicking a benign giant cell tumor . A 22-year-old male was admitted to our hospital with a history of mild left wrist pain after a skiing trauma . Radiology revealed an extensive meta-epiphyseal osteolytic lesion in the distal ulna , which appeared to be a giant cell tumor . Histological examination showed a biphasic tumor comprising chondroid and non-chondroid areas with a giant cell-rich lesion resembling a conventional giant cell tumor of the bone . Immunohistochemistry showed no expression of p16(INK4a) , P17948 , P35968 ( P35968 ) , P35916 , cKIT , Q00987 or P11802 . However , high expression of the tyrosine kinases P16234 and P09619 was observed . Molecular analysis showed no amplification of the cMYC gene and no activating mutations in the cKIT ( exons 9 and 11 ) or P16234 ( exon 18 ) genes . He has been on follow-up for ten months , with no evidence of local recurrence or metastatic disease . In summary , this report highlights a very rare case of a dedifferentiated chondrosarcoma in which the dedifferentiated component of the tumor bears histologic resemblance to a conventional giant cell tumor of bone . We suggest that this tumor might be categorized in the group of low-grade dedifferentiated chondrosarcomas . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Boswellic acid exerts antitumor effects in colorectal cancer cells by modulating expression of the let-7 and miR-200 microRNA family . Colorectal cancer ( CRC ) is a complex disease with genetic and epigenetic alterations in many key oncogenes and tumor suppressor genes . The active principle of a gum resin from Boswellia serrata , 3-acetyl-11-keto-β-boswellic acid ( AKBA ) , has recently gained attention as a chemopreventive compound due to its ability to target key oncogenic proteins such as P09917 and nuclear factor-kappaB . AKBA has been shown to inhibit the growth of CRC cells ; however , the precise molecular mechanisms underlying its anticancer activities in CRC remain unclear . We hypothesized that boswellic acids may achieve their chemopreventive effects by modulating specific microRNA ( miRNA ) pathways . We found that AKBA significantly up-regulated expression of the let-7 and miR-200 families in various CRC cell lines . Both let-7 and miR-200 are putative tumor-suppressive miRNAs . AKBA modulated the expression of several downstream targets of the let-7 and miR-200 families , such as Q00534 , vimentin and P12830 . These data were further strengthened by miRNA knockdown studies , which revealed that inhibition of let-7i facilitated enhanced cancer cell proliferation , migration and invasion . In addition , AKBA also induced similar modulation of the let-7 and miR-200 downstream genes in CRC tumors orthotopically implanted in nude mice . These results indicate that AKBA-induced antitumor effects in CRC occur , at least partly through the up-regulation of specific miRNA pathways . Our data provide novel evidence that anticancer effects of boswellic acids are due in part to their ability to regulate cellular epigenetic machinery and further highlight the promise for this phytochemical in the preventative and therapeutic applications of CRC . Protein kinase C activation and the development of diabetic complications . Recent studies have identified that the activation of protein kinase C ( PKC ) and increased diacylglycerol ( DAG ) levels initiated by hyperglycemia are associated with many vascular abnormalities in retinal , renal , and cardiovascular tissues . Among the various PKC isoforms , the beta- and delta-isoforms appear to be activated preferentially in the vasculatures of diabetic animals , although other PKC isoforms are also increased in the renal glomeruli and retina . The glucose-induced activation of PKC has been shown to increase the production of extracellular matrix and cytokines ; to enhance contractility , permeability , and vascular cell proliferation ; to induce the activation of cytosolic phospholipase A2 ; and to inhibit Na+-K+-ATPase . The synthesis and characterization of a specific inhibitor for P05771 isoforms have confirmed the role of PKC activation in mediating hyperglycemic effects on vascular cells , as described above , and provide in vivo evidence that PKC activation could be responsible for abnormal retinal and renal hemodynamics in diabetic animals . Transgenic mice overexpressing P05771 isoform in the myocardium developed cardiac hypertrophy and failure , further supporting the hypothesis that P05771 isoform activation can cause vascular dysfunctions . Interestingly , hyperglycemia-induced oxidative stress may also mediate the adverse effects of P05771 isoforms by the activation of the DAG-PKC pathway , since treatment with D- DB00163 was able to prevent many glucose-induced vascular dysfunctions and inhibit DAG-PKC activation . Clinical studies are now in progress to determine whether P05771 inhibition can prevent diabetic complications . New isoflavonoids as inhibitors of porcine P09917 . The inhibitory activity of new isoflavonoids on P09917 of porcine leukocytes was investigated . Isoflavans ( I ) proved to be stronger inhibitors than isoflavones ( II ) . The isoflavans containing ortho-hydroxy groups in ring A showed the lowest Ki values ( 0.8-50 microM ) . In comparison , isoflavans with meta-dihydroxy groups exhibited Ki values higher than 150 microM . The effect of commercial antioxidants was tested also on porcine P09917 . Butylated hydroxyanisole ( Ki : 25 microM ) and butylated hydroxytoluene ( Ki : 55 microM ) revealed moderate inhibitory activity , whereas L-ascorbic acid , L-ascorbyl palmitate , dl- DB00163 and n-propyl gallate showed weak inhibitory activities ( Ki : 100-260 microM ) . Alpha-tocopherol decreases superoxide anion release in human monocytes under hyperglycemic conditions via inhibition of protein kinase C-alpha . Diabetes is a major risk factor for premature atherosclerosis , and oxidative stress appears to be an important mechanism . Previously , we showed that diabetic monocytes produce increased superoxide anion ( O(2)(-) ) , and DB00163 ( AT ) supplementation decreases this . The aim of this study was to elucidate the mechanism(s) of O(2)(-) release and inhibition by AT under hyperglycemic ( HG ) conditions in monocytes . O(2)(-) release , protein kinase C ( PKC ) activity , and translocation of P17252 and -betaII and p47phox were increased in THP-1 cells ( human monocytic cell line ) under HG ( 15 mmol/l glucose ) conditions , whereas AT supplementation inhibited these changes . AT , NADPH oxidase inhibitors ( apocynin and diphenyleneiodonium chloride [ DPI ] ) , and an inhibitor to P17252 and other isoforms ( 2,2',3,3',4,4'-hexahydroxy-1,1'-biphenyl-6,6'-dimethanol dimethyl ether [ HBDDE ] ) but not P05771 II ( LY379196 ) decreased O(2)(-) release and p47phox translocation . Antisense oligodeoxynucleotides to P17252 and p47phox but not to PKC-betaII inhibited HG-induced O(2)(-) release and p47phox translocation in THP-1 cells . Under HG conditions , reactive oxygen species release from monocytes was not inhibited by agents affecting mitochondrial metabolism but was inhibited in human endothelial cells . We conclude that under HG conditions , monocytic O(2)(-) release is dependent on NADPH oxidase activity but not the mitochondrial respiratory chain ; HG-induced O(2)(-) release is triggered by P17252 , and AT inhibits O(2)(-) release via inhibition of P17252 . The effect of DB00163 on monocyte proatherogenic activity . Atherosclerosis is the leading cause of morbidity and mortality in Westernized populations . The monocyte is a crucial cell in the genesis of the atherosclerotic lesion and is present during all stages of atherosclerosis . alpha-Tocopherol ( AT ) is the most active component of the vitamin E family and is the principal and most potent lipid-soluble antioxidant in plasma and LDL . With regard to monocyte function , AT supplementation ( 1200 IU/d ) has been shown to decrease release of reactive oxygen species , lipid oxidation , release of cytokines such as interleukin-1ss ( IL-1ss ) and tumor necrosis factor-alpha ( P01375 ) and decrease adhesion of monocytes to human endothelium . The mechanism of inhibition of superoxide and lipid oxidation by monocytes appears to be via inhibition of protein kinase C ( PKC ) , the decrease in IL-1ss and P01375 release by inhibition of P09917 and the inhibition of monocyte-endothelial cell adhesion via decrease in adhesion molecules on monocytes , CD11b and VLA-4 and by decreasing DNA-binding activity of nuclear transcription factor kappaB . Thus , in addition to the decrease in oxidative stress resulting from AT supplementation , as evidenced by decreased F(2)-isoprostanes and LDL oxidizability , AT is anti-inflammatory and exerts beneficial antiatherogenic effects on cells crucial in atherogenesis such as monocytes . DB00163 prevents diabetes-induced abnormal retinal blood flow via the diacylglycerol-protein kinase C pathway . We have characterized effects of d- DB00163 ( vitamin E ) on activation of protein kinase C ( PKC ) and diacylglycerol ( DAG ) levels in retinal tissues of diabetic rats and correlated its effects to diabetes-induced changes in retinal hemodynamics . Membrane PKC specific activities were increased by 71 % in streptozocin-induced diabetic rats compared with controls ( P < 0.05 ) . Western blot analysis showed that membrane P05771 II was increased by 133 +/- 5 % ( P < 0.05 ) . Injection of d- DB00163 ( 40 mg/kg ip ) every other day prevented the increases in membrane PKC specific activity and P05771 II protein by immunoblots . Diabetes-induced increases in DAG levels were also normalized by d- DB00163 treatment of 2 wk duration . Physiologically , angiographic abnormalities of retinal hemodynamics based on computerized video-based fluorescein angiography and associated with increases of DAG and membranous PKC levels were also prevented by d- DB00163 treatment in diabetic rats . The effect of d- DB00163 on retinal vascular cells was also studied . Exposure of retinal endothelial cells to 22 mM glucose for 3 days increased total DAG and [3H]palmitate-labeled DAG levels by 35 +/- 8 and 50 +/- 8 % ( P < 0.05 ) , respectively , compared with exposure to 5.5 mM glucose . The presence of d- DB00163 ( 50 micrograms/ml ) prevented the increases in total DAG and [3H]palmitate-labeled DAG levels in cells exposed to 22 mM glucose . These findings suggested that treatment with d- DB00163 can prevent diabetes-induced abnormalities in rat retinal blood flow. ( ABSTRACT TRUNCATED AT 250 WORDS ) Oxidized LDL and lysophosphatidylcholine stimulate plasminogen activator inhibitor-1 expression through reactive oxygen species generation and P27361 /2 activation in 3T3- Q9NUQ9 adipocytes . P00747 activator inhibitor-1 ( P05121 ) is secreted from adipose tissue and is considered to be a risk factor for both atherosclerosis and insulin resistance . Here we report for the first time that P05121 expression is enhanced by oxidized low-density lipoprotein ( OxLDL ) and its lipid component lysophosphatidylcholine ( Q16549 ) in mouse 3T3- Q9NUQ9 adipocytes . In fully differentiated 3T3- Q9NUQ9 cells , OxLDL treatment increased the mRNA expression and protein secretion of P05121 in a dose- and time-dependent manner , whereas native LDL had no effect . The addition of an anti- P16671 antibody suppressed OxLDL-stimulated P05121 expression by 50 % , suggesting that adipose-derived P16671 contributes to roughly half of the P05121 expression stimulated by OxLDL . In addition , pharmacological experiments showed that the OxLDL-stimulated enhancement in P05121 expression was mediated through the generation of reactive oxygen species ( ROS ) and phosphorylation of extracellular signal-regulated kinase 1/2 . Furthermore , Q16549 , a major lipid component of OxLDL , was responsible for the enhanced expression of P05121 as phospholipase A(2)-treated acetyl LDL , which generates Q16549 , strongly stimulated P05121 expression , whereas acetyl LDL itself had no such activity . These data demonstrate that the uptake of OxLDL and , in particular , its lipid component Q16549 into adipocytes triggers aberrant ROS-mediated P05121 expression , which may be involved in the pathogenesis of metabolic syndrome . DB00163 suppresses P09917 -mediated oxidative stress in peripheral blood mononuclear cells of hemodialysis patients regardless of administration route . A number of pathological conditions caused by oxidative stress have been reported in uremic patients undergoing maintenance hemodialysis ( HD ) . Enhanced lipid peroxidation was previously observed in peripheral blood mononuclear cells ( PBMCs ) of HD patients . Upregulation of P09917 ( 5-Lox ) activity and protein content with enhanced production of leukotriene B(4) ( Q06643 (4) ) and membrane lipoperoxides was also shown in PBMCs of HD patients . Administration of free vitamin E specifically inhibited 5-Lox activity without affecting gene expression at the protein level . To assess whether oral or intramuscular ( IM ) administration of vitamin E may suppress 5-Lox in HD patients , PBMCs from 16 subjects on maintenance HD therapy for at least 6 months were investigated before and after a short course of IM or oral administration of vitamin E ( 8 patients per group ) . PBMCs from 13 healthy controls were also evaluated and assumed as the reference standard . DB00163 significantly reduced lipid peroxidation , Q06643 (4) content , and 5-Lox activity in PBMCs , whereas 5-Lox gene expression at the protein level was not affected . There were no significant differences in these parameters between patients treated with IM or oral vitamin E. PBMCs of HD patients showed enhanced membrane lipid peroxidation and release of Q06643 (4) , both linked to upregulation of 5- P28300 : 5-Lox activity and related oxidative stress were significantly ( although not completely ) suppressed by vitamin E regardless of the administration route . Design , synthesis , and biological evaluation of conformationally constrained aci-reductone mimics of arachidonic acid . An efficient and convergent synthesis has been developed for the production of 3,4-dihydroxy-5- [ 4- ( 2- ( ( 2Z ) -hexenyl ) phenyl ) -3-(1Z)-but enyl ] -2 ( 5H ) -furanone ( 12d ) . This hydrophobic antioxidant is a stable conformationally constrained mimic of arachidonic acid ( AA ) ( 1 ) and its respective aci-reductone analogue ( 2 ) . Pd(0)-catalyzed cross-coupling of 5-(3-butynyl)-3,4-dihydroxy-2(5H)-furanone ( 7 ) with 2- ( ( 2Z ) -hexenyl ) iodobenzene ( 8d ) followed by Lindlar catalyzed hydrogenation produces 12d . Butynyl intermediate 7 is prepared from 2-(benzyloxy)-5-deoxyascorbic acid ( 15 ) by iodination ( I2 , PPh3 , Imd ) , iodo substitution with lithium acetylide ethylenediamine complex ( LiAEDA , HMPA , -5 degrees C ) , and benzyl group cleavage ( Ac2O , Pyr , BCl3 ) . The utility of this synthetic method was demonstrated by the synthesis of analogues 10e-k . Biological testing revealed that certain of these antioxidants inhibit both cyclooxygenase ( P36551 ) and P09917 ( P09917 ) with comparable efficacy as reported for aspirin and zileuton , respectively . The antioxidant activity of these aci-reductones , measured as a function of their inhibitory effect on CCl4-induced lipid peroxidation of hepatic microsomes , exceeds that produced by DB00163 . Synthetic routes and initial structure-activity relationships ( SAR ) for these novel mixed functioning antioxidants are presented . Some antioxidants inhibit , in a co-ordinate fashion , the production of tumor necrosis factor-alpha , IL-beta , and P05231 by human peripheral blood mononuclear cells . Some antioxidants , including butylated hydroxyanisole ( BHA ) , tetrahydropapaveroline ( THP ) , nordihydroguiauretic acid , and 10,11-dihydroxyaporphine ( DB01708 ) , were found to be potent inhibitors of the production of tumor necrosis factor ( P01375 ) -alpha , P01584 , and P05231 by human peripheral blood mononuclear cells ( PBMC ) stimulated by lipopolysaccharide ( LPS ) ( IC50s in the low micromolar range ) . Inhibition of cytokine production was gene selective and not due to general effects on protein synthesis . Inhibition of cytokine production by PBMC was observed also when other inducers were used ( staphylococci , silica , zymosan ) . Much higher concentrations of other antioxidants -- including ascorbic acid , trolox , DB00163 , butylated hydroxytoluene , and the P09917 inhibitor zileuton -- did not affect the production of these cytokines . The active compounds did not inhibit IL-1-induced production of P05231 in fibroblasts , showing the cell selectivity of the effect . Antioxidant-mediated inhibition of cytokine production was correlated with low levels of the corresponding messenger RNAs . Nuclear run-on experiments showed that THP inhibited transcription of the P01584 gene . THP decreased the concentration of the transcription factors NF-kappa B and AP-1 detected in nuclear extracts of PBMC cultured in the presence or absence of LPS . THP and DB01708 markedly decreased the levels of P01375 and P01584 in the circulation of mice following LPS injection . Thus antioxidants vary widely in potency as inhibitors of the activation of transcription factors and of the transcription of genes for pro-inflammatory cytokines . Coordinate inhibition of the transcription of genes for inflammatory cytokines could provide a strategy for therapy of diseases with inflammatory pathogenesis and for septic shock . Specific cellular responses to DB00163 . In the last 10 years precise cellular functions of DB00163 , some of which are independent of its antioxidant/radical-scavenging ability , have been revealed . Absorption of DB00163 from the gut is a selective process . Other tocopherols are not absorbed or are absorbed to a lesser extent . At the post-translational level , DB00163 inhibits protein kinase C and P09917 and activates protein phosphatase 2A and diacylglycerol kinase . Some genes [ platelet glycoprotein IV/thrombospondin receptor/class B scavenger receptor ( P16671 ) , DB00163 transfer protein ( alpha-TTP ) , alpha-tropomyosin , connective tissue growth factor and collagenase ] are affected by DB00163 at the transcriptional level . alpha-Tocopherol also inhibits cell proliferation , platelet aggregation , monocyte adhesion and the oxygen burst in neutrophils . Other antioxidants , such as beta-tocopherol and probucol , do not mimic these effects , suggesting a nonantioxidant , DB00163 -specific molecular mechanism . Activation of PKC but not of P29323 is required for vitamin E-succinate-induced apoptosis of HL-60 cells . DB00163 -succinate ( VES ) induced HL-60 human leukemia cells to undergo apoptosis . Treatment with VES induced membrane translocation of Fas ; cleavages of caspase-3 , PARP , and lamin B ; hypophosphorylation of retinoblastoma protein ; and increase of P38936 ( P38936 ) protein level . During the induction of apoptosis , activity of PKC was gradually increased with downregulation of VES-induced P29323 activity and accompanied by activation of caspase-3 . Inhibition of PKC by GF109203X blocked VES-mediated membrane translocation of P17252 and cleavage of caspase-3 cascade , resulting in prevention of VES-induced apoptosis . On the contrary , PKC activation by cotreatment with Q16549 or thapsigargin and VES synergistically increased VES-mediated apoptosis . However , inhibition of P29323 activity by PD98059 showed no significant effect on VES-induced PKC activity and apoptosis . Taken together , our data suggest that VES induces activation of PKC and PKC-dependent hypophosphorylation of retinoblastoma protein , which results in induction of apoptosis , and that VES-induced early activation of P29323 and P29323 -dependent induction of P38936 ( P38936 ) are not required for apoptosis . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . Activation of P09917 and related cell membrane lipoperoxidation in hemodialysis patients . Lipid peroxidation was shown at the membrane level in peripheral blood cells of patients hemodialyzed on cuprophan dialyzers , and was mainly attributable to the generation of conjugated hydroperoxides in the lipid bilayer . The oxidative index ( i.e. , the A234/205 ratio ) of membrane lipids was 3.2-fold higher in hemodialysis patients than in healthy control subjects , and also the level of leukotriene B4 was significantly increased ( up to 1.7-fold over control ) . Both membrane peroxidation and release of leukotriene B4 were linked to upregulation of P09917 activity ( up to 2.4-fold over control ) and expression at the protein level ( up to 1.9-fold ) . DB00163 , the most important lipophilic antioxidant , prevented both membrane peroxidation and release of leukotriene B4 by inhibiting P09917 activity without affecting enzyme expression . Similar results were observed in patients hemodialyzed on polymethylmetacrylate membranes , but in this case the activation of P09917 was less pronounced . The use of a purified P09917 demonstrated that vitamin E was a reversible inhibitor of enzyme activity ( IC50 = 35 +/- 4 microM ) , further characterized as noncompetitive ( Ki = 30 +/- 3 microM ) . Taken together , the results reported here shed some light on the mechanism responsible for the oxidative damage in hemodialysis . Moreover , the beneficial effect of vitamin E described here may have relevance for the therapy of patients with kidney disease . Effect of NC-1900 , an active fragment analog of arginine vasopressin , and inhibitors of arachidonic acid metabolism on performance of a passive avoidance task in mice . In this study , we investigated the effect of administration of inhibitors of each of the arachidonic acid metabolism pathways and the effect of co-administration of these inhibitors with NC-1900 , a fragment analog of arginine vasopressin , on step-through passive avoidance task performance . All drugs were administered just after the acquisition trial in the passive avoidance task . Intracerebroventricular ( i.c.v. ) administration of nordihydroguaiaretic acid ( NDGA , 1 and 10 microg ) , a phospholipase A2 ( P04054 ) and lipoxygenase ( P28300 ) inhibitor , and of arachidonyl trifluoromethyl ketone ( Q06187 , 1 and 10 microg ) , a specific P04054 inhibitor caused reductions in latency on the retention trial . The i.c.v. administration of either of baicalein ( 0.1-10 microg ) , a P16050 inhibitor , or AA-861 ( 0.1-10 microg ) , a 5- P28300 inhibitor , did not influence the latency . Intraperitoneal administration of indomethacin ( 20 mg/kg ) , a non-specific P36551 inhibitor , or NS-398 ( 10 mg/kg ) , a specific P35354 inhibitor , impaired performance on the retention trial in the task , while piroxicam ( 20 mg/kg ) , a specific P23219 inhibitor , did not . Subcutaneous administration of NC-1900 ( 0.1 ng/kg ) ameliorated the reduction of latency caused by NDGA , Q06187 , indomethacin , or NS-398 . These results suggested that the P35354 pathway of arachidonic acid metabolism may be important for learning and/or memory in the passive avoidance task in mice , and that the ameliorating effect of NC-1900 , in part , is due to mimicking of the effects of metabolites of the P35354 pathway . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Smad6s regulates plasminogen activator inhibitor-1 through a protein kinase C-beta-dependent up-regulation of transforming growth factor-beta . P00747 activator inhibitor-1 ( P05121 ) is a serpin class protease inhibitor that plays a central role in the regulation of vascular function and tissue remodeling by modulating thrombosis , inflammation , and the extracellular matrix . A central mediator controlling P05121 is transforming growth factor-beta ( TGF-beta ) , which induces its expression and promotes fibrosis . We have found that a unique member of the Smad family of signal transduction molecules , Smad6s , modulates the expression of P05121 . Overexpression of Smad6s in endothelial cells increases promoter activity and P05121 secretion , and an antisense to Smad6s suppresses the induction of P05121 by TGF-beta . The effect of Smad6s on the P05121 promoter appeared to be the result of increase binding of the forkhead winged helix factor FoxD1 to a TGF-beta-responsive element . Furthermore , the effect of Smad6s on P05121 up-regulation and on FoxD1 binding was found to result from up-regulation of TGF-beta and could be inhibited by the blocking TGF-beta signaling with O15105 . The ability of Smad6s to regulate the TGF-beta promoter and subsequent P05121 induction was suppressed by a selective protein kinase C-beta ( P05771 ) inhibitor . Consistent with the in vitro data , we found that increased Smad6s in diseased vessels correlated with increased TGF-beta and P05121 levels . Overall , our results demonstrate that the level of Smad6s can alter the level of TGF-beta and the subsequent induction of P05121 via a FoxD1 transcription site . Furthermore , our data suggest that this process , which is up-regulated in diseased vessels , can be modulated by the inhibition of P05771 . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Effects of dietary vitamin E on the biosynthesis of P09917 products by rat polymorphonuclear leukocytes ( PMNL ) . Activation of polymorphonuclear neutrophils ( PMNL ) leads to the release of arachidonate from cellular phospholipids via a phospholipase A2 , and conversion of products of the P09917 pathway . Evidence to date indicates the dietary vitamin E ( ( R,R,R ) - DB00163 ) can influence both cyclooxygenase and phospholipase A2 activities and that the effect of this vitamin is cell/tissue specific . The present study was undertaken in order to examine the effects of varying dietary tocopherol on PMNL tocopherol content and P09917 product profile using the ionophore A23187 as stimulant in the presence and absence of exogenous arachidonate . Feeding semi-purified diets containing 0 , 30 or 3000 ppm of ( R,R,R ) - DB00163 acetate to weanling rats for 17 weeks resulted in a dose-related enrichment of PMNL tocopherol . Stimulation of PMNL elicited a significant and rapid loss of tocopherol . When PMNL were stimulated with A23187 alone , the synthesis of 5-HETE , LTB4 and 19-hydroxy-LTB4 was decreased in proportion to increasing dietary tocopherol concentrations . However , when exogenous arachidonate was provided with A23187 , intermediate amounts of dietary tocopherol ( 30 ppm ) still suppressed the formation of P09917 products , but high doses ( 3000 ppm ) did not have any additional inhibitory effect . This differential response to high concentrations of vitamin E in the presence and absence of exogenous arachidonate highly suggest that at these concentrations , tocopherol may act principally at the level of substrate release whereas at lower concentrations , P09917 is inhibited . Data from this study demonstrated that attenuation of the formation of P09917 products in PMNL can be achieved by dietary vitamin E enrichment . Exploring the roles of P22309 and P35503 in oral clearance of GSK2190915 , a P09917 -activating protein inhibitor . Pharmacokinetic variability in drug exposure is a concern for all compounds in development including those for the treatment of asthma and other respiratory disorders . Substantial variability in the oral clearance of GSK2190915 , a P09917 -activating protein inhibitor that attenuates the production of leukotriene B4 and cysteinyl leukotrienes , is largely unaccounted for by clinical variables . A study of 41 patients , 78 % ( 32/41 ) of whom were non-Hispanic whites , with mild to moderate asthma identified an association of P22309 *28 and P35503 *2 with the oral clearance of GSK2190915 ( P=3.8×10⁻⁴ and 1.2×10⁻⁵ , respectively ) . However , in a subsequent replication study of 403 non-Hispanic white patients with asthma , we failed to observe a statistically significant association between oral clearance of GSK2190915 and either P22309 *28 or P35503 *2 ( P > 0.05 ) . Therefore , genetic effects that could explain the systemic exposure level variability of GSK2190915 were not identified . Reduction of human monocytic cell neurotoxicity and cytokine secretion by ligands of the cannabinoid-type CB2 receptor . 1 Two cannabinoid receptors , P21554 and CB2 , have been identified . The P21554 receptor is preferentially expressed in brain , and the CB2 receptor in cells of leukocyte lineage . We identified the mRNA for the P21554 receptor in human neuroblastoma SH-SY5Y cells , and the mRNA and protein for the CB2 receptor in human microglia and THP-1 cells . 2 Delta(9)-and Delta(8)-tetrahydrocannabinol ( THC ) were toxic when added directly to SH-SY5Y neuroblastoma cells . The toxicity of Delta(9)- THC was inhibited by the P21554 receptor antagonist SR141716A but not by the CB2 receptor antagonist SR144528 . The endogenous ligand anandamide was also toxic , and this toxicity was enhanced by inhibitors of its enzymatic hydrolysis . 3 The selective CB2 receptor ligands JWH-015 and indomethacin morpholinylamide ( BML-190 ) , when added to THP-1 cells before stimulation with lipopolysaccharide ( LPS ) and P01579 , reduced the toxicity of their culture supernatants to SH-SY5Y cells . JWH-015 was more effective against neurotoxicity of human microglia than THP-1 cells . The antineurotoxic activity of JWH-015 was blocked by the selective CB2 receptor antagonist SR144528 , but not by the P21554 receptor antagonist SR141716A . This activity of JWH-015 was synergistic with that of the P09917 ( 5- P28300 ) inhibitor REV 5901 . 4 Cannabinoids inhibited secretion of IL-1beta and tumor necrosis factor-alpha ( P01375 ) by stimulated THP-1 cells , but these effects could not be directly correlated with their antineurotoxic activity . 5 Specific CB2 receptor ligands could be useful anti-inflammatory agents , while avoiding the neurotoxic and psychoactive effects of P21554 receptor ligands such as Delta(9)-THC . Inhibition of P09917 by vitamin E . Purified P09917 from potato tubers was inhibited strongly by vitamin E and its analogs . The inhibition by d- DB00163 was found to be irreversible and non-competitive with respect to arachidonic acid . An IC50 of 5 microM was calculated for d- DB00163 . The inhibition appears to be unrelated to its antioxidant function . Binding studies with 14C-labelled d- DB00163 revealed that there is a strong interaction between vitamin E and P09917 . Tryptic digestion and peptide mapping of P09917 -vitamin E complex indicate that vitamin E binds strongly to a single peptide . These studies suggest that cellular vitamin E levels may have profound influence on the formation of leukotrienes . Gamma-tocopherol , but not DB00163 , decreases proinflammatory eicosanoids and inflammation damage in rats . Gamma-tocopherol ( gammaT ) , the major form of vitamin E in U.S. diets , and its physiological metabolite 2 , 7 , 8-trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman ( gamma-CEHC ) , in contrast to DB00163 ( alphaT ) , the primary vitamin E in supplements , inhibit cyclooxygenase-catalyzed synthesis of prostaglandin E2 ( DB00917 ) in activated macrophages and epithelial cells . Here we report that in carrageenan-induced inflammation in male Wistar rats , administration of gammaT ( 33 or 100 mg/kg ) and gamma-CEHC ( 2 mg/pouch ) , but not alphaT ( 33 mg/kg ) , significantly reduced DB00917 synthesis at the site of inflammation . gammaT , but not alphaT , significantly inhibited the formation of leukotriene B4 , a potent chemotactic agent synthesized by the P09917 of neutrophils . Although gammaT had no effect on neutrophil infiltration , it significantly attenuated the partial loss of food consumption caused by inflammation-associated discomfort . Administration of gammaT led consistently to a significant reduction of inflammation-mediated increase in 8-isoprostane , a biomarker of lipid peroxidation . gammaT at 100 mg/kg reduced P01375 ( 65 % ;P=0.069 ) , total nitrate/nitrite ( 40 % ;P=0.1 ) , and lactate dehydrogenase activity ( 30 % ;P=0.067 ) . Collectively , gammaT inhibits proinflammatory DB00917 and LTB4 , decreases P01375 , and attenuates inflammation-mediated damage . These findings provide strong evidence that gammaT shows anti-inflammatory activities in vivo that may be important for human disease prevention and therapy . Prolonged effects of tumor necrosis factor-alpha on anterior pituitary hormone release . We examined the chronic ( 72 h ) effects of 30 ng/ml recombinant murine tumor necrosis factor ( P01375 ) -alpha on release of immunoreactive growth hormone ( GH ) , prolactin ( PRL ) , thyrotropin ( DB00024 ) , and DB00024 glycosylation , as assessed by lectin binding , in cultured rat anterior pituitary cells . In cultured cells from adult female rats , P01375 significantly suppressed basal and GH-releasing hormone ( P01148 ) -stimulated GH release . P01375 also suppressed basal PRL release and completely abolished the PRL response to TRH ( 0.1-10 nM ) . Whereas P01375 reduced basal DB00024 release , it significantly enhanced the maximal DB00024 response to TRH . P01375 did not affect the concanavalin A and lentil lectin binding of DB00024 accumulated in the medium during the 4-day culture , but significantly decreased the lentil lectin binding of DB00024 released in response to acute TRH stimulation . P01375 significantly enhanced the inhibitory effect of somatostatin on stimulated PRL release , but not on GH or DB00024 release . Compared to cell cultures from adult female rats , in anterior pituitary cell cultures from 12-day-old rats the effects of prolonged exposure to P01375 on hormone release were diminished or absent . Pituitary hormone release was unaffected by acute ( 3 h ) exposure to P01375 . These results demonstrate a direct effect of P01375 on anterior pituitary hormone release , which is cell-type specific and age dependent . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Differential neurotrophic regulation of sodium and calcium channels in an adult sympathetic neuron . Adult neuronal phenotype is maintained , at least in part , by the sensitivity of individual neurons to a specific selection of neurotrophic factors and the availability of such factors in the neurons ' environment . Nerve growth factor ( P01138 ) increases the functional expression of Na(+) channel currents ( I(Na) ) and both N- and L-type Ca(2+) currents ( I(Ca,N) and I(Ca,L) ) in adult bullfrog sympathetic ganglion ( BFSG ) B-neurons . The effects of P01138 on I(Ca) involve the mitogen-activated protein kinase ( MAPK ) pathway . Prolonged exposure to the ganglionic neurotransmitter luteinizing hormone releasing hormone ( P01148 ) also increases I(Ca,N) but the transduction mechanism remains to be elucidated as does the transduction mechanism for P01138 regulation of Na(+) channels . We therefore exposed cultured BFSG B-neurons to chicken II P01148 ( 0.45 microM ; 6-9 days ) or to P01138 ( 200 ng/ml ; 9-10 days ) and used whole cell recording , immunoblot analysis , and ras or rap-1 pulldown assays to study effects of various inhibitors and activators of transduction pathways . We found that 1 ) P01148 signals via ras-MAPK to increase I(Ca,N) , 2 ) this effect is mediated via protein kinase C-beta ( P05771 -IotaIota ) , 3 ) protein kinase A ( PKA ) is necessary but not sufficient to effect transduction , 4 ) P01138 signals via phosphatidylinositol 3-kinase ( PI3K ) to increase I(Na) , and 5 ) long-term exposure to P01148 fails to affect I(Na) . Thus downstream signaling from P01148 has access to the ras-MAPK pathway but not to the PI3K pathway . This allows for differential retrograde and anterograde neurotrophic regulation of sodium and calcium channels in an adult sympathetic neuron . Population pharmacokinetic study of memantine : effects of clinical and genetic factors . BACKGROUND AND OBJECTIVE : Memantine , a frequently prescribed anti-dementia drug , is mainly eliminated unchanged by the kidneys , partly via tubular secretion . Considerable inter-individual variability in plasma concentrations has been reported . We aimed to investigate clinical and genetic factors influencing memantine disposition . METHODS : A population pharmacokinetic study was performed including data from 108 patients recruited in a naturalistic setting . Patients were genotyped for common polymorphisms in renal cation transporters ( O15245 /2/5 , Q96FL8 , P08183 ) and nuclear receptors ( O75469 , Q14994 , RXR , Q07869 ) involved in transporter expression . RESULTS : The average clearance was 5.2 L/h with a 27 % inter-individual variability ( percentage coefficient of variation ) . Glomerular filtration rate ( p = 0.007 ) and sex ( p = 0.001 ) markedly influenced memantine clearance . O75469 rs1523130 was identified as the unique significant genetic covariate for memantine clearance ( p = 0.006 ) , with carriers of the O75469 rs1523130 CT/TT genotypes presenting a 16 % slower memantine elimination than carriers of the CC genotype . CONCLUSION : The better understanding of inter-individual variability of memantine disposition might be beneficial in the context of individual dose optimization . Splenic marginal zone lymphoma : proposal of new diagnostic and prognostic markers identified after tissue and cDNA microarray analysis . Splenic marginal zone lymphoma ( SMZL ) is a newly recognized lymphoma type whose precise molecular pathogenesis is still essentially unknown . This hampers differential diagnosis with other small B-cell malignancies . With the aim of characterizing this tumor more comprehensively , and of identifying new diagnostic and prognostic markers , we performed cDNA microarray expression profiling and tissue microarray ( TMA ) immunohistochemical studies in a relatively large series of 44 SMZLs . The results were related to immunoglobulin heavy chain variable region ( IgV(H) ) mutational status and clinical outcome . SMZLs display a largely homogenous signature , implying the existence of a single molecular entity . Of the genes deregulated in SMZLs , special mention may be made of the genes involved in B-cell receptor ( P11274 ) signaling , tumor necrosis factor ( P01375 ) signaling and nuclear factor-kappaB ( NF-kappaB ) activation , such as P43405 , Q06187 , Q13489 , Q13114 , and Q06643 . Other genes observed were P14151 and O60711 , which were highly expressed in spleen , and lymphoma oncogenes , such as Q15669 and TCL1 . In contrast , the genes Q03135 , P51636 , and P61952 located in 7q31 , a commonly deleted area , were down-regulated in the entire series . A comparison with the genes comprising the signature of other small B-cell lymphomas identified 3 genes whose expression distinguishes SMZL , namely Q01167 , SENATAXIN , and P25942 . Shorter survival was associated with P28907 expression , naive IgV(H) genes , and the expression of a set of NF-kappaB pathway genes , including O00463 , Q04864 , and P17252 . The role of protein kinase C activation in the pathogenesis of diabetic vascular complications . Many vascular diseases in diabetes are known to be associated with the activation of the diacylglycerol ( DAG ) -protein kinase C ( PKC ) pathway . The major source of DAG that is elevated in diabetes is de novo synthesis from glycolytic intermediates . Among the various PKC isoforms , the beta-isoform has been shown to be persistently activated in diabetic animals . Multiple lines of evidence have shown that many vascular alterations in diabetes -- such as a decrease in the activity of Na+-K+-adenosine triphosphatase ( Na+-K+-ATPase ) , and increases in extracellular matrix , cytokines , permeability , contractility , and cell proliferation -- are caused by activation of PKC . Inhibition of PKC by two different kinds of PKC inhibitors , LY333531 , a selective P05771 -isoform inhibitor , and d- DB00163 , were able to prevent or reverse the various vascular dysfunctions in diabetic rats . These results have also provided in vivo evidence that DAG-PKC activation could be responsible for the hyperglycemia-induced vascular dysfunctions in diabetes . Clinical studies are now being performed to clarify the pathogenic roles of the DAG-PKC pathway in developing vascular complications in diabetic patients . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Periadventitial adipose tissue impairs coronary endothelial function via P05771 -dependent phosphorylation of nitric oxide synthase . Endogenous periadventitial adipose-derived factors have been shown to contribute to coronary vascular regulation by impairing endothelial function through a direct inhibition of endothelial nitric oxide synthase ( P29474 ) . However , our understanding of the underlying mechanisms remains uncertain . Accordingly , this study was designed to test the hypothesis that periadventitial adipose tissue releases agents that attenuate coronary endothelial nitric oxide production via a protein kinase C ( PKC ) -beta-dependent mechanism . Isometric tension studies were conducted on isolated canine circumflex coronary arteries with and without natural amounts of periadventitial adipose tissue . Adipose tissue significantly diminished coronary endothelial-dependent vasodilation and nitric oxide production in response to bradykinin and acetylcholine . The selective inhibition of endothelial P05771 with ruboxistaurin ( 1 microM ) abolished the adipose-induced impairment of bradykinin-mediated coronary vasodilation and the endothelial production of nitric oxide . Western blot analysis revealed a significant increase in P29474 phosphorylation at the inhibitory residue DB00156 (495) in arteries exposed to periadventitial adipose tissue . This site-specific phosphorylation of P29474 was prevented by the inhibition of P05771 . These data demonstrate that periadventitial adipose-derived factors impair coronary endothelial nitric oxide production via a P05771 -dependent , site-specific phosphorylation of P29474 at DB00156 (495) . Luteinizing Hormone-Releasing Hormone ( P01148 ) -I antagonist cetrorelix inhibits myeloma cell growth in vitro and in vivo . The objective of this study was to determine the effects of an luteinizing hormone-releasing hormone ( P01148 ) -I antagonist , DB00050 , on human multiple myeloma ( MM ) cells and to elucidate the mechanisms of action . We showed that P01148 -I and P22888 -I genes were expressed in MM cell lines and primary MM cells . Treatment with DB00050 inhibited growth and colony-forming ability of myeloma cells , including cell lines resistant to arsenic trioxide , bortezomib , or lenalidomide . DB00050 induced apoptosis in myeloma cells including primary myeloma cells . In addition , DB00050 inhibited the growth of human myeloma cells xenografted into mice without any apparent side effects . DB00050 downregulated the nuclear factor-kappa B ( NF-κB ) pathway activity and the expression of cytokines , including interleukin 6 , insulin-like growth factor 1 , P15692 , and stromal-derived factor 1 , important for myeloma cell growth and survival in myeloma cells and/or marrow stromal cells from myeloma patients . DB00050 decreased the phosphorylation of extracellular signal regulated kinase 1/2 and P40763 in myeloma cells , two crucial pathways for myeloma cells growth and survival . Moreover , the expression of P38936 and p53 was increased , whereas that of antiapoptotic proteins Bcl-2 and Bcl-x(L) was reduced by DB00050 . Our findings indicate that DB00050 induces cytotoxicity in myeloma cells through various mechanisms and provide a rationale for investigating DB00050 for the treatment of MM . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 .

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Novel path to P05231 trans-signaling through thrombin-induced soluble P05231 receptor release by platelets . Interleukin ( IL ) -6 is a multifunctional cytokine with a critical role in inflammatory , immunoregulatory and haemopoietic responses . Its receptor consists of an ubiquitously expressed membrane transducing element ( P40189 ) and of the specific element P08887 ( P08887 ) , present only on hepatocytes and some leukocyte subsets . P08887 also exists as soluble protein ( sIL-6R ) that , in the presence of P05231 , forms a complex able to bind P40189 and , thanks to the mechanism called trans-signaling , transduces P05231 effect through tyrosine phosphorylation and activation of the signal transducer and transcription activator ( P35610 ) -3 . The aim of this study was to analyze the bidirectional relationships between platelet aggregation and P05231 -dependent effects . While platelets do not produce P05231 , we found that resting platelets express P40189 , but not P08887 , on their membranes . Upon activation by thrombin or calcium ionophore A23187 , but not by ADP , the P08887 is released in soluble form , while cangrelor , the specific inhibitor of Q9H244 receptor , can partially inhibit sIL-6R release . This sIL-6R is biologically active and , in the presence of P05231 , can trigger P05231 trans-signaling , inducing an autocrine activation loop ( as measured by an increase in P08887 and P40189 content ) and P40763 phosphorylation . On the other hand , P05231 trans-signaling has no effect on platelet degranulation or aggregation by itself , nor on thrombin-induced platelet aggregation . Our data add an important piece to the puzzle of thrombosis and inflammation : in the presence of P05231 , which can be produced by stressed endothelial cells , the platelet-derived P05231 trans-signaling could be crucial for the evolution of inflammation within a damaged vessel . State of the art of new Q9H244 antagonists . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : ( a ) it is a pro-drug that needs to be metabolized to its active metabolite ; ( b ) it has a delayed onset and offset of action and ( c ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and a more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG-related bleeding complications . Two direct and reversible Q9H244 antagonists , DB06441 and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 is not superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 is superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but , like prasugrel , is associated with a higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery is necessary in ticagrelor-treated patients compared to clopiodgrel-treated patients to limit the severity of post-surgical bleeding . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . DB06441 : review of the drug and the CHAMPION programme ( including PHOENIX ) . Platelet inhibition is the main goal of ancillary pharmacologic therapy during percutaneous coronary interventions ( P05154 ) . Thienopyridines and ticagrelor are oral drugs developed for this purpose . DB06441 is an intravenous , non-thienopyridine antagonist of the Q9H244 receptor with a rapid , potent , predictable , and quickly reversible effect . DB06441 has been studied in a broad population intended to receive P05154 in the CHAMPION program , where it was compared with different clopidogrel regimens . The first two trials , CHAMPION P05154 and PLATFORM , failed their primary objective , likely for challenges in the adjudication of P05154 -related myocardial infarction . In a third trial that implemented the universal definition of MI , CHAMPION PHOENIX , a reduction of thrombotic events , including stent thrombosis , was observed . In the BRIDGE trial cangrelor has been studied in patients who had to prematurely interrupt antiplatelet therapy for surgery . DB06441 appears a promising agent in patients who require P05154 or when a rapid reversal is needed . Transitioning patients from cangrelor to clopidogrel : pharmacodynamic evidence of a competitive effect . BACKGROUND : DB06441 is a direct , parenteral , and reversible inhibitor of the platelet Q9H244 receptor currently undergoing Phase III testing . As many individuals treated acutely with cangrelor will often be treated long-term with a thienopyridine , it is important to determine the effects of concurrent cangrelor and clopidogrel administration . METHODS AND RESULTS : Ten healthy volunteers received a 600 mg oral loading dose of clopidogrel and then underwent serial platelet function monitoring for 6 h . Two weeks later these same individuals received a 600 mg clopidogrel loading dose simultaneously with a cangrelor IV bolus ( 30 microg/kg ) and a 2-hour infusion ( 4 microg/kg/min ) . A separate group of ten volunteers received a 600 mg clopidogrel loading dose after administration of a cangrelor bolus and a 1-hour infusion . The effects on ADP-induced platelet activation and aggregation were evaluated by flow cytometry , whole-blood electrical impedance , and light-transmittance aggregometry . DB06441 and clopidogrel alone achieved the expected levels of platelet inhibition . However , the sustained platelet inhibition anticipated for clopidogrel treatment did not occur when cangrelor was initiated simultaneously . No such effect was found when clopidogrel was started upon completion of the cangrelor infusion . CONCLUSION : To achieve sustained platelet Q9H244 inhibition in patients treated with cangrelor , clopidogrel administration should be started when the cangrelor infusion is terminated . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . Purine receptor Q15077 mediates cellular response to γ-ray-induced DNA damage . We previously showed that nucleotide P2 receptor agonists such as DB00171 and UTP amplify γ-ray-induced focus formation of phosphorylated histone H2A variant P16104 ( γ P16104 ) , which is considered to be an indicator of DNA damage so far , by activating purine Q15077 and Q9H244 receptors . Therefore , we hypothesized that these P2 receptors play a role in inducing the repair response to γ-ray-induced DNA damage . In the present study , we tested this idea by using human lung cancer A549 cells . First , reverse-transcription polymerase chain reaction ( RT-PCR ) showed that Q15077 receptor is highly expressed in A549 cells , but Q9H244 receptor is only weakly expressed . Next , colony formation assay revealed that Q15077 receptor antagonist MRS2578 markedly reduced the survival rate of γ-ray-exposed A549 cells . The survival rate was also significantly reduced in Q15077 -knock-down cells , compared with scramble siRNA-transfected cells . Since it has reported that phosphorylation of P27361 /2 after activation of P00533 via Q15077 and Q9H244 receptors is involved in the repair response to γ-ray-induced DNA damage , we next examined whether γ-ray-induced phosphorylation of P27361 /2 was also inhibited by MRS2578 in A549 cells . We found that it was . Taken together , these findings indicate that purinergic signaling through Q15077 receptor , followed by P27361 /2 activation , promotes the cellular repair response to γ-ray-induced DNA damage . New frontiers in the management of acute coronary syndromes : cangrelor and elinogrel . The activation and aggregation of platelets at sites of vascular injury or near to implanted stent are pivotal in the development of thrombotic events during and after an acute coronary syndrome ( ACS ) or a percutaneous coronary intervention ( P05154 ) . For that reason , an exclusively oral dual antiplatelet treatment regimen with platelet Q9H244 receptor antagonists in addition to the cyclooxygenase inhibitor aspirin has become the cornerstone of treatment in that contest . However , every trial underlines the same problem : if maximizing antiplatelet therapy significantly attenuates ischemic events in patients with coronary artery disease , on the other side it may also increase bleeding phenomena . These limitations have prompted a search for novel antiplatelet agents with a more favorable risk-benefit ratio . Moreover , an early onset of action is desirable during P05154 and an early offset after bleeding events . Two novel antiplatelet agents , DB06441 and Elinogrel , are available in intravenous form ( Elinogrel also in oral form ) and expand this context . Recent trials have tested them against DB00758 regarding efficacy and safety outcomes.This review aimed at providing an overview on intravenous emerging compounds and recent patents in the setting of ACS and P05154 . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . DB06441 : a review on pharmacology and clinical trial development . Dual antiplatelet therapy with aspirin and an oral ADP Q9H244 receptor antagonist is the standard-of-care for the prevention of ischemic events in patients with acute coronary syndrome or undergoing percutaneous coronary intervention ( P05154 ) . However , currently available ADP Q9H244 receptor antagonists have several limitations , such as interindividual response variability , drug-drug interactions , slow onset/offset and only oral availability . DB06441 is a reversible , potent , intravenous , competitive inhibitor of the ADP Q9H244 receptor that rapidly achieves near complete and predictable platelet inhibition . Along with reversible binding to the receptor cangrelor also has a very short half-life ( 3-5 min ) , which in turn results in a rapid offset of action . These properties make cangrelor a promising drug for clinical use in patients undergoing P05154 or patients waiting for major surgery but still require antiplatelet protection . This manuscript provides an update of the current status of knowledge on cangrelor , focusing on its pharmacologic properties and clinical trial development , including the BRIDGE and CHAMPION-PHOENIX trials . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . DB00619 inhibition effect on KITAsn822Lys-mediated signal transduction cascade . OBJECTIVE : Alterations in growth factor signaling pathways may be a frequent collaborating event in Q01196 - Q06455 -mediated leukemogenesis . Gain-of-function P10721 receptor mutations have been reported in adult AML patients , especially those with core binding factor leukemia ( CBFL ) . We have previously reported a new gain-of-function P10721 (Asn822Lys) mutation that is constitutively expressed in the Kasumi-1 CBFL cell line , and has recently been described in two childhood AML patients . To explore the molecular basis of the effects of this mutation in the appropriate context of hemopoietic dysregulation , we investigated P10721 downstream signaling in the Kasumi-1 cell line by means of DB00619 ( Imatinib , Gleevec ) pharmacological inhibition . MATERIALS AND METHODS : We investigated P10721 (Asn822Lys) mutant-initiated signaling in Kasumi-1 cell line , and characterized the inhibitory effect of the DB00619 protein tyrosine kinase inhibitor on downstream signaling . RESULTS : The use of DB00619 -mediated inhibition impaired the tyrosine phosphorylation of P10721 (Asn822Lys) and its association with the p85 subunit of phosphatidylinositol 3'-kinase ( p85PI3K ) . The downstream constitutive phosphorylation of P45983 /2 and P40763 was also significantly inhibited , but DB00619 had no effect on the constitutive activation of Akt , thus suggesting that it is due to other signaling in Kasumi-1 cells . DB00619 inhibited the P10721 -mediated proliferation of Kasumi-1 cells in a dose-dependent manner . CONCLUSIONS : These findings show the role of PI3K in P10721 (Asn822Lys)-mediated constitutive activation through the Akt-independent downstream signaling pathway of JNK , and also demonstrate the mutant 's susceptibility to DB00619 , which may therefore have therapeutic potential in CBFL patients with susceptible P10721 mutations . DB06441 : a novel Q9H244 receptor antagonist . Antiplatelet therapy is critical in the prevention of thrombotic complications of acute coronary syndrome and percutaneous coronary interventions . Current antiplatelet agents ( aspirin , clopidogrel and glycoprotein IIb/IIIa antagonists ) have demonstrated the capacity to reduce major adverse cardiac events . However , these agents have limitations that compromise their clinical utility . The platelet Q9H244 receptor plays a central role in platelet function and is a focus in the development of antiplatelet therapies . DB06441 is a potent , competitive inhibitor of the Q9H244 receptor that is administered by intravenous infusion and rapidly achieves near complete inhibition of ADP-induced platelet aggregation . This investigational drug has been studied for use during coronary procedures and the management of patients experiencing acute coronary syndrome and is undergoing evaluation for use in the prevention of perioperative stent thrombosis . P42892 and β-arrestins exert spatiotemporal control of DB05875 -induced inflammatory signals . Although the intracellular trafficking of G protein-coupled receptors controls specific signaling events , it is unclear how the spatiotemporal control of signaling contributes to complex pathophysiological processes such as inflammation . By using bioluminescence resonance energy transfer and superresolution microscopy , we found that DB05875 ( SP ) induces the association of the neurokinin 1 receptor ( P25103 ) with two classes of proteins that regulate SP signaling from plasma and endosomal membranes : the scaffolding proteins β-arrestin ( βARRs ) 1 and 2 and the transmembrane metallopeptidases ECE-1c and ECE-1d . In HEK293 cells and non-transformed human colonocytes , we observed that G protein-coupled receptor kinase 2 and β P49407 /2 terminate plasma membrane Ca(2+) signaling and initiate receptor trafficking to endosomes that is necessary for sustained activation of ERKs in the nucleus . βARRs deliver the SP- P25103 endosomes , where P42892 associates with the complex , degrades SP , and allows the P25103 , freed from βARRs , to recycle . Thus , both P42892 and βARRs mediate the resensitization of P25103 Ca(2+) signaling at the plasma membrane . Sustained exposure of colonocytes to SP activates NF-κB and stimulates P10145 secretion . This proinflammatory signaling is unaffected by inhibition of the endosomal P29323 pathway but is suppressed by P42892 inhibition or β P32121 knockdown . Inhibition of protein phosphatase 2A , which also contributes to sustained P25103 signaling at the plasma membrane , similarly attenuates P10145 secretion . Thus , the primary function of βARRs and P42892 in SP-dependent inflammatory signaling is to promote resensitization , which allows the sustained P25103 signaling from the plasma membrane that drives inflammation . Central Q9H244 receptor blockade alleviates inflammatory and neuropathic pain and cytokine production in rodents . In this study the role of Q9H244 receptors ( P2Y12R ) was explored in rodent models of inflammatory and neuropathic pain and in acute thermal nociception . In correlation with their activity to block the recombinant human P2Y12R , the majority of P2Y12R antagonists alleviated mechanical hyperalgesia dose-dependently , following intraplantar O75347 injection , and after partial ligation of the sciatic nerve in rats . They also caused an increase in thermal nociceptive threshold in the hot plate test . Among the six P2Y12R antagonists evaluated in the pain studies , the selective Q9H244 receptor antagonist PSB-0739 was most potent upon intrathecal application . P2Y12R mRNA and IL-1β protein were time-dependently overexpressed in the rat hind paw and lumbar spinal cord following intraplantar O75347 injection . This was accompanied by the upregulation of P01375 -α , P05231 and P22301 in the hind paw . PSB-0739 ( 0.3mg/kg i.t. ) attenuated O75347 -induced expression of cytokines in the hind paw and of IL-1β in the spinal cord . Subdiaphragmatic vagotomy and the α7 nicotinic acetylcholine receptor antagonist MLA occluded the effect of PSB-0739 ( i.t. ) on pain behavior and peripheral cytokine induction . Denervation of sympathetic nerves by 6-OHDA pretreatment did not affect the action of PSB-0739 . PSB-0739 , in an analgesic dose , did not influence motor coordination and platelet aggregation . Genetic deletion of the P2Y12R in mice reproduced the effect of P2Y12R antagonists on mechanical hyperalgesia in inflammatory and neuropathic pain models , on acute thermal nociception and on the induction of spinal IL-1β . Here we report the robust involvement of the P2Y12R in inflammatory pain . The anti-hyperalgesic effect of P2Y12R antagonism could be mediated by the inhibition of both central and peripheral cytokine production and involves α7-receptor mediated efferent pathways . [ Recent advances on the studies of the platelet 's inhibition and aggregation. State of the art of new Q9H244 antagonists ] . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : i ) it is a pro-drug that needs to be metabolized to its active metabolite ; ii ) it has a delayed onset and offset of action ; iii ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG ( Coronary Artery Bypass Grafting ) related bleeding complications . Two direct and reversible Q9H244 antagonists , cangrelor and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 did not prove superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 proved to be superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but was , like prasugrel , was associated with higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery was necessary in ticagrelor-treated patients compared to clopidogrel-treated patients to limit the severity of post-surgical bleeding . Contribution of the Q9H244 receptor-mediated pathway to platelet hyperreactivity in hypercholesterolemia . BACKGROUND : In hypercholesterolemia , platelets demonstrate increased reactivity and promote the development of cardiovascular disease . OBJECTIVE : This study was carried out to investigate the contribution of the ADP receptor Q9H244 -mediated pathway to platelet hyperreactivity due to hypercholesterolemia . METHODS : P01130 -deficient mice and C57Bl/6 wild-type mice were fed on normal chow and high-fat ( Western or Paigen ) diets for 8 weeks to generate differently elevated cholesterol levels . Q9H244 receptor-induced functional responses via G(i) signaling were studied ex vivo when washed murine platelets were activated by 2MeSADP and PAR4 agonist AYPGKF in the presence and absence of indomethacin . Platelet aggregation and secretion , α(IIb)β(3) receptor activation and the phosphorylation of extracellular signal-regulated protein kinase ( P29323 ) and Akt were analyzed . RESULTS : Plasma cholesterol levels ranged from 69 ± 10 to 1011 ± 185 mg dL(-1) depending on diet in mice with different genotypes . Agonist-dependent aggregation , dense and α-granule secretion and JON/A binding were gradually and significantly ( P < 0.05 ) augmented at low agonist concentration in correlation with the increasing plasma cholesterol levels , even if elevated thromboxane generation was blocked . These functional responses were induced via increased levels of G(i) -mediated P29323 and Akt phosphorylation in hypercholesterolemic mice vs. normocholesterolemic animals . In addition , blocking of the Q9H244 receptor by AR-C69931MX ( DB06441 ) resulted in strongly reduced platelet aggregation in mice with elevated cholesterol levels compared with normocholesterolemic controls . CONCLUSIONS : These data revealed that the Q9H244 receptor pathway was substantially involved in platelet hyperreactivity associated with mild and severe hypercholesterolemia . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . P01308 / P05019 signaling pathways enhances tumor cell invasion through bisecting GlcNAc N-glycans modulation . an interplay with P12830 . Changes in glycosylation are considered a hallmark of cancer , and one of the key targets of glycosylation modifications is P12830 . We and others have previously demonstrated that P12830 has a role in the regulation of bisecting GlcNAc N-glycans expression , remaining to be determined the P12830 -dependent signaling pathway involved in this N-glycans expression regulation . In this study , we analysed the impact of P12830 expression in the activation profile of receptor tyrosine kinases such as insulin receptor ( IR ) and P08069 ( IGF-IR ) . We demonstrated that exogenous P12830 expression inhibits IR , IGF-IR and P29323 1/2 phosphorylation . Stimulation with insulin and P05019 in MDA-MD-435 cancer cells overexpressing P12830 induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on P12830 cellular localization . Concomitantly , IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability . Altogether , these results demonstrate an interplay between P12830 and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression , with important effects in the modulation of epithelial characteristics and tumor cell invasion . Here we provide new insights into the role that P01308 / P05019 signaling play during cancer progression through glycosylation modifications . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . New Q9H244 blockers . A number of new antiplatelet agents currently in development are anticipated to improve clinical outcomes and safety benefits in patients with acute coronary syndrome ( ACS ) . This article reviews the pharmacology and clinical development of three of these agents : prasugrel , cangrelor , and ticagrelor . Prasugrel , a third-generation , oral thienopyridine , has been shown to be superior to clopidogrel , the current gold standard , in preventing ischemic events in patients with ACS undergoing percutaneous coronary intervention ( P05154 ) , although the bleeding rate was higher . DB06441 , a chemical analog of adenosine triphosphate , is a potent direct platelet Q9H244 antagonist . In development as an intravenous agent , cangrelor is currently being evaluated in two phase III studies in patients requiring P05154 . DB08816 is the first of a new class of orally available antiplatelet agents antagonizing the effects of ADP mediated by Q9H244 ; it is currently being studied in a phase III trial in patients with ACS . DB06441 for treatment of arterial thrombosis . INTRODUCTION : Percutaneous coronary intervention ( P05154 ) is a highly effective treatment for obstructive coronary artery disease . Oral platelet Q9H244 receptor antagonists reduce ischemic events in patients treated with P05154 . However , there are several limitations to their use , including variable pharmacodynamics , a slow onset and offset , and in those patients who are pretreated but subsequently require cardiac surgery , increased bleeding . DB06441 is an intravenous agent that provides rapid and intensive inhibition of the Q9H244 receptor that quickly dissipates after discontinuation . A recent , Phase III randomized clinical trial of P05154 patients demonstrated that cangrelor bolus and infusion reduced ischemic events compared with conventional clopidogrel therapy without increasing major bleeding . AREAS COVERED : This review outlines the pharmacodynamics , pharmacokinetics , and the safety and efficacy of cangrelor for the acute treatment of patients undergoing planned P05154 . EXPERT OPINION : DB06441 is an important addition to the current armamentarium of platelet inhibitors as it significantly reduces periprocedural myocardial infarction and stent thrombosis in a broad spectrum of patients , without increasing major bleeding or the need for transfusion . DB06441 will have particular benefit in clopidogrel-naïve patients with high anatomical complexity and/or increased clinical risk ( where the absolute risk for thrombotic and ischemic complications of P05154 is greatest ) . Knockdown of receptor-interacting serine/threonine protein kinase-2 ( O43353 ) affects EMT-associated gene expression in human hepatoma cells . BACKGROUND : Receptor-interacting serine/threonine protein kinase-2 ( O43353 ) has been reported to be an important regulator of tumor proliferation , differentiation and wound repair . We investigated the effects of O43353 knockdown in human hepatoma cells on epithelial-to-mesenchymal transition ( EMT ) -associated gene expression . MATERIALS AND METHODS : HepG2 cells stably expressing O43353 -shRNA ( HepG2-shRIPK2 ) were generated after puromycin selection . Total RNAs from HepG2-shRIPK2 and from HepG2-shcontrol cells were isolated and PCR-based arrays were performed to compare the 84 EMT-associated gene expressions . RESULTS : We observed that knockdown of O43353 down-regulated mRNA expression of jagged 1 ( P78504 ) ; plasminogen activator inhibitor-1 ( P05121 ) ; regulator of G-protein signalling 2 , 24 kDa ( P41220 ) ; P12830 ( CDH1 ) ; fibroblast growth factor binding protein 1 ( Q14512 ) ; snail homolog 2 ( O43623 ) ; protein tyrosine phosphatase type IVA , member 1 ( Q93096 ) ; keratin 19 ( P08727 ) ; vimentin ( P08670 ) ; and survival of motor neuron protein-interacting protein 1 ( SIP1 ) . CONCLUSION : We found that knockdown of O43353 down-regulated nuclear factor kappa B ( NF-κB ) -dependent P05121 and P08670 gene expressions . O43353 might play an important role in hepatic cell migration . These findings could shed new light on carcinogenesis and on liver regeneration . DB03843 - or adjuvant-induced peripheral inflammation increases neurokinin-1 receptor gene expression in the mouse . Substance P ( SP ) has been widely studied as a mediator of nociception . The release of SP from primary afferent neurons is increased during nociception , and SP activates neurokinin-1 ( NK-1 ) receptors in the spinal cord and periphery . Nociception-evoked alterations in P25103 gene expression have been studied in rat models of persistent pain but have not been characterized in any murine models of peripheral inflammation . This study assessed behavioral responses and P25103 mRNA gene expression in mice receiving formalin or Freund 's complete adjuvant ( O75347 ) as an inflammatory stimulus . Mechanical withdrawal thresholds were measured before injection of formalin or O75347 and hind paw licking/biting timed during the late-phase of the formalin response . Two and 24 hours after formalin or O75347 injection , mechanical withdrawal thresholds were measured and the mice euthanized . Solution hybridization-nuclease protection assays were used to quantify P25103 mRNA levels . Results demonstrated that inflamed hind paws were edematous , and the withdrawal thresholds of the inflamed hind paws were significantly lower after formalin or O75347 injection . Neurokinin-1 receptor mRNA levels in the ipsilateral dorsal spinal cords of mice were higher at 24 h after formalin injection or 4 days after O75347 injection . These results confirm that mice are hyperalgesic at late time points after formalin or adjuvant injection when P25103 gene expression is elevated in the dorsal spinal cord . This supports the hypothesis that increased P25103 gene expression contributes to the development and maintenance of a hyperalgesic state . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Interaction of murine peritoneal leukocytes and mesothelial cells : in vitro model system to survey cellular events on serosal membranes during inflammation . All serosal cavities including peritoneum are lined with a simple squamous mesothelium . Primary culture of murine mesothelial cells has been established to study their cellular interactions with peritoneal leukocytes . The mesothelial character was determined by the cytokeratin and vimentin expression . The mesothelial cells expressed P05362 and P16070 molecules . The expression of P05362 , but not P16070 , was significantly enhanced by the treatment with P01375 ( 100 U/ml ) . We have also investigated possible influence of transforming growth factors , TGF-alpha ( 20 ng/ml ) and TGF-beta ( 2 ng/ml ) , and epidermal growth factor ( 20 ng/ml ) . These factors were not found to modulate P05362 or P16070 expression in vitro . During coculture experiments unstimulated mesothelial cells were almost nonadherent for both resident and elicited peritoneal mononuclear leukocytes for several hours . P01375 or P01133 pretreatment of mesothelial cells greatly enhanced their adhesive affinity to peritoneal mononuclear leukocytes , while TGF-beta pretreatment even reduced the low basal adhesion . Prolonged coculture for 3 weeks resulted in remarkable proliferation and differentiation of both resident and elicited monocytes/macrophages on the mesothelial surface . The stimulation of mesothelial cell culture with P01133 resulted in the macrophage colony-stimulating activity ( M- Q13216 ) production . M- Q13216 was mainly due to P09603 as confirmed with anti P09603 monoclonal antibody ; the residual M- Q13216 was not formed by GM- P04141 . After several passages the mesothelial cells started to produce M- Q13216 spontaneously . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . DB06441 in percutaneous coronary intervention . DB06441 is a novel , intravenous Q9H244 receptor antagonist in development for use in percutaneous coronary intervention . Currently in Phase III testing , the reversible platelet inhibitor provides several inherent advantages over other Q9H244 receptor antagonists in this setting for the prevention of adverse cardiac events . Unlike the class of thienopyridines ( ticlopidine , clopidogrel and potentially soon to be available , prasugrel ) , cangrelor has nearly immediate onset after a bolus dose and a short half-life , and achieves maximal inhibition of ADP-mediated platelet function . DB06441 's distinct mechanism of action allows for intravenous administration and avoids both hepatic and renal metabolism . These unique characteristics make cangrelor a promising agent for use in cardiovascular patients undergoing percutaneous coronary intervention . Purinergic receptors involved in the immunomodulatory effects of DB00171 in human blood . We recently showed that the physiological compound DB00171 simultaneously inhibited P01375 and stimulated P22301 release in LPS-PHA stimulated blood . The purpose of the present study was to determine the mechanism involved in the concerted modulatory effect of DB00171 on P01375 and P22301 . Incubation of blood with DB00171 in the presence of selective P2 receptor antagonists showed that the stimulatory effect of DB00171 on P22301 release was completely annihilated by both 2-MeSAMP ( a Q9H244 /13 receptor antagonist ) and PSB-0413 ( a Q9H244 receptor antagonist ) . On the other hand , the inhibitory effect of DB00171 on P01375 release was completely reversed by 5'- P30566 ( a Q96G91 receptor antagonist ) as well as by H-89 , an inhibitor of DB02527 -activated PKA . The concerted inhibition by DB00171 of P01375 release via Q96G91 activation and stimulation of P22301 release via Q9H244 activation implicates a novel approach towards immunomodulation by altering the balance among pro- and anti-inflammatory cytokines . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Pharmacokinetics and pharmacodynamics of a bolus and infusion of cangrelor : a direct , parenteral Q9H244 receptor antagonist . The purpose of this study is to evaluate the safety , tolerability , pharmacokinetics , and pharmacodynamics of cangrelor administered as an intravenous bolus plus a continuous infusion in healthy volunteers . Twenty-two healthy volunteers are randomized to receive 1 of 2 intravenous cangrelor dosing regimens : a 15-microg/kg bolus followed by a 2-microg/kg/min infusion or a 30-microg/kg bolus followed by a 4-microg/kg/min infusion . The infusion is continued for 60 minutes , and serial blood samples are obtained for evaluation of pharmacokinetic and pharmacodynamic parameters . Administration of an intravenous bolus followed by a continuous infusion rapidly achieves maximum concentrations of cangrelor that are associated with extensive platelet inhibition within 2 minutes . Moreover , extensive platelet inhibition is maintained throughout the infusion period with near-full recovery of platelet function within 60 to 90 minutes of terminating the infusion . The effect of high-dose cangrelor is more consistent and demonstrates a greater level of inhibition on adenosine diphosphate-induced P16109 expression ; how ever , no significant differences are observed between the 2 dosing regimens with regard to platelet aggregation or time to recovery of platelet function . DB06441 administered as an intravenous bolus followed by a continuous infusion in healthy volunteers offers rapid and reversible inhibition of platelet function . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone .

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interacts_with DB00831?

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In vivo silencing of aquaporin-1 by RNA interference inhibits angiogenesis in the chick embryo chorioallantoic membrane assay . P29972 ( P29972 ) is a water channel protein mainly expressed in endothelial and epithelial cells of many tissues , including the vasculature where it serves to increase cell membrane water permeability . Previous studies in active multiple myeloma patients and in P29972 KO mice indicated an involvement of P29972 in physiological and tumor angiogenesis . To understand the physiological role of P29972 in angiogenesis , we used a 21-nucleotide small interfering RNA duplexes ( siRNA ) to knockdown P29972 in the chick embryo chorioallantoic membrane ( P62158 ) , a commonly used in vivo assay to study both angiogenic and angiostatic molecules . Chicken P29972 sequence was identified and utilized to synthesize a siRNA directed to the P29972 sequence . We then tested the efficiency of the siRNA in vitro , using an P29972 transfected cell line . The level of P29972 protein reduction obtained using siRNA was 98 % and 92 % after 1 and 2 day transfection respectively . RNA interference experiments were then performed in vivo by using the P62158 assay . Results showed that after 4 days of treatment , P29972 siRNA was able to strongly inhibit angiogenesis . This is the first study showing the in vivo use of RNA interference technique in the P62158 assay . Our results strongly support the hypothesis that P29972 could have a key role in physiological and pathological angiogenesis . Skinned coronary smooth muscle : calmodulin , calcium antagonists , and DB02527 influence contractility . The effects of Ca2+ , calmodulin , DB02527 , the catalytic subunit of DB02527 -dependent protein kinase ( CSU ) and some Ca2+ antagonists were studied in chemically ( Triton X-100 ) skinned coronary smooth muscle . P62158 increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration . DB00831 , a calmodulin antagonist , inhibited Ca2+-calmodulin-induced contraction . Both DB02527 and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration . DB08980 , a Ca2+-antagonist , at 2 x 10(-4) M produced a significant inhibitory effect , which was reduced by increasing the Ca2+ concentration . From other Ca2+ antagonists tested , W-7 , but not D600 and verapamil , produced some inhibitory effect . The data indicate that the response of skinned coronary smooth muscle to Ca2+ , calmodulin and DB02527 are similar to those obtained with other skinned smooth muscles . Furthermore , skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle . Gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides by cDNA microarrays . To investigate the gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides ( ODNs ) , Q15761 antisense , mismatched ODNs or vehicle was intracerebroventricularly injected and cDNA microarrays were undertaken . Central administration of Q15761 antisense ODNs decreased food intake , body weight and serum insulin compared with both vehicle and mismatched ODNs . The average area of adipocytes both at retroperitoneal and epididymal adipose tissue were fall in antisense group while only the weight of the retroperitoneal fat pats was reduced in antisense group . cDNA microarrays containing 18,000 genes/Ests were used to investigate gene expression of adipose tissue . Autoradiographic analysis showed that 404 , 81 , and 34 genes were differently expressed over twofold , threefold , and fivefold , respectively . The analysis of gene expression profiles indicated that 332 genes were up-regulated and 187 genes were down-regulated in response to Q15761 antisense ODNs treatment . Different clusters of genes associated with apoptosis , signal transduction , energy metabolism , lipid metabolism , etc. , such as P51114 , Q8WV24 , Q7L5Y9 , P27986 , P13598 , Q00169 , P62158 , Q13557 , P61925 , P14416 , O95258 , CKB , P22760 , P38571 , O15254 , O60427 , were concerned . Analysis of differentially expressed genes will help to understand the effects of Q15761 antisense ODNs therapy . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . [ P62158 -dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium ] . Highly purified plasma membrane ( PM ) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein . Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3 % of the original level . The activity of Ca , Mg-ATPase is thereby decreased by 40 % . Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour . The maximal activation of Ca , Mg-ATPase is observed with 10(-7) M calmodulin . P62158 increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity . DB00831 ( 20 microM ) diminishes the activating effect of exogenous calmodulin on Ca , Mg-ATPase . P62158 stimulates Ca , Mg-ATPase at low concentrations of Ca2+ ( 10(-8)-10(-6) M ) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax -- from 0,8 to 1.42 mumol Pl/mg protein/hour . It is supposed that the activating effect of calmodulin on Ca , Mg-ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . Signaling cascade that mediates endothelial nitric oxide synthase activation induced by atrial natriuretic peptide . Atrial natriuretic peptide ( P01160 ) induces activation of nitric oxide-synthase ( NOS ) . AIMS : to identify the isoform of NOS involved in P01160 effects , to study whether P01160 modifies NOS expression and to investigate the signaling pathways and receptors involved in NOS stimulation . NOS activation induced by P01160 would be mediated by endothelial NOS ( P29474 ) since neuronal or inducible NOS inhibition did not alter P01160 effect . The peptide induced no changes in P29474 protein expression . NOS activity stimulated by P01160 , in the kidney , aorta and left ventricle , was partially abolished by the P16066 /B antagonist , as well as PKG inhibition , but no difference in atria was observed . 8-Br-cGMP partially mimicked the effect of P01160 on NOS in all tissues . NOS stimulation by P01160 in atria disappeared when G protein was inhibited , but this effect was partial in the other tissues . P62158 antagonist abolished NOS stimulation via P01160 . Inhibition of the P98160 , PKC or P19957 kinase/Akt pathway failed to alter NOS activation induced by P01160 . P01160 would activate P29474 in the aorta , heart and kidney without modifying the expression of the enzyme . P01160 would interact with P17342 coupled via G proteins leading to the activation of Ca(2+)-calmodulin-dependent NOS in atria ; while in ventricle , aorta and kidney , P01160 could also interact with P16066 /B , increasing cGMP , which in turns activates PKG to stimulate P29474 . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Epitope-specific antibody response to Mel- P62158 induced by mimotope immunization . Peptide mimotopes of tumor antigen epitopes have been proposed as components of tumor vaccines . In this study , we determined the immunogenicity of melcam mim1 and melcam mim2 , peptide mimics of an epitope of the melanoma cell-adhesion molecule ( Mel- P62158 ) . BALB/c mice were vaccinated either with mimotopes or mimotopes coupled to tetanus toxoid ( TT ) . The antibody responses of mice to melcam mim1 , melcam mim2 , and recombinant Mel- P62158 were analyzed by an ELISA and immunoblot analyses . TT-coupled mimotopes led to high titers of IgG mainly of the IgG2a subclass to melcam mim1 and melcam mim2 . Immunization with each of the mimotope formulations induced antibodies that cross-reacted with recombinant Mel- P62158 . Uncoupled mimotopes induced lymphocyte proliferation and cytokine production in spleen cell cultures indicating that both peptide mimotopes also contained T cell epitopes . TT-coupled mimotopes induced T helper (Th)1 ( interleukin ( IL ) -2 , interferon-gamma ) and Th2 ( P05112 , P05113 ) cytokines , whereas uncoupled mimotopes induced a Th1-biased T cell response . Our results suggest that mimotopes potentially represent a novel vaccine approach to induce a tumor antigen-specific humoral and cellular response . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Establishment and phenotypic characterization of human U937 cells with inducible P210 P11274 / P00519 expression reveals upregulation of P13688 ( CD66a ) . Chronic myeloid leukemia ( CML ) is characterized by the expression of the P210 P11274 / P00519 fusion protein . The molecular mechanisms behind this oncogene-mediated hematological disease are , however , not fully understood . Here , we describe the establishment and phenotypic characterization of U937 cells in which P210 P11274 / P00519 can be conditionally expressed using tetracycline . The induction of P11274 / P00519 in the obtained clones resulted in a rapid phosphorylation of the P42224 , P40763 and P42229 molecules , consistent with the findings in other model systems . Phenotypic characterization of the clones revealed that P11274 / P00519 induces a slight decrease in the proliferation and viability , without a marked effect on cell cycle distribution , the rate of apoptosis or on cellular differentiation , as judged by several cell surface markers and capacity to reduce nitro blue tetrazolium . Interestingly , P11274 / P00519 was found to upregulate the expression of carcinoembryonic-related antigen ( P06731 ) P62158 ( CD66a ) , which is a plasma membrane-linked glycoprotein belonging to the CEAs and involved in signal transduction and cellular adhesion . The expression of P13688 was reversible upon imatinib treatment in P11274 / P00519 -expressing U937 cells as well as in P11274 / P00519 -positive K562 cells . The established cell lines may prove useful in further modeling and dissection of P11274 / P00519 -induced leukemogenesis . Effects of pulsed electromagnetic fields on human osteoblastlike cells ( MG-63 ) : a pilot study . BACKGROUND : Although pulsed electromagnetic fields ( PEMFs ) are used to treat delayed unions and nonunions , their mechanisms of action are not completely clear . However , PEMFs are known to affect the expression of certain genes . QUESTIONS/PURPOSES : We asked ( 1 ) whether PEMFs affect gene expression in human osteoblastlike cells ( MG63 ) in vitro , and ( 2 ) whether and to what extent stimulation by PEMFs induce cell proliferation and differentiation in MG-63 cultures . METHODS : We cultured two groups of MG63 cells . One group was treated with PEMFs for 18 hours whereas the second was maintained in the same culture condition without PEMFs ( control ) . Gene expression was evaluated throughout cDNA microarray analysis containing 19,000 genes spanning a substantial fraction of the human genome . RESULTS : PEMFs induced the upregulation of important genes related to bone formation ( P31260 , P31749 ) , genes at the transductional level ( P62158 , Q99572 ) , genes for cytoskeletal components ( P02751 , P18206 ) , and collagenous ( P08123 ) and noncollagenous ( P09486 ) matrix components . However , PEMF induced downregulation of genes related to the degradation of extracellular matrix ( P24347 , Q13115 ) . CONCLUSIONS AND CLINICAL RELEVANCE : PEMFs appear to induce cell proliferation and differentiation . Furthermore , PEMFs promote extracellular matrix production and mineralization while decreasing matrix degradation and absorption . Our data suggest specific mechanisms of the observed clinical effect of PEMFs , and thus specific approaches for use in regenerative medicine . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Mite antigens enhance P05362 and induce P19320 expression on human umbilical vein endothelium . Although sublingual allergen-specific immunotherapy has been proved to be effective in the treatment of allergic diseases , controversy surrounds the means by which such a local therapy can induce systemic immunological changes . Adhesion molecules are critical in the regulation of leukocyte traffic . It has been hypothesized that allergenic extract , administered locally , may induce an up-regulation of the mucosal vessel vascular adhesion molecules ( CAMs ) resulting in local recruitment of circulating inflammatory cells . In the present study we investigated whether the mite antigens , Der p1 and Der p2 , can modulate P62158 expression of human endothelial cells ( O14777 ) . To do this , slices of whole human umbilical cord vein underwent short-term ( 8 hours ) cultures in the presence or absence of mite antigen ( baseline , unstimulated controls ) . Cryostatic sections of the specimens were then evaluated immunohistochemically for expression of intercellular adhesion molecule ( P05362 ) and vascular cell adhesion molecule ( P19320 ) molecules . The results revealed that while Der p1 is capable of significantly up-regulating P05362 and P19320 on O14777 , Der p2 antigen moderately up-regulates P05362 expression but is ineffective in modulating P19320 . Although preliminary , these results clearly support the hypothesis that at least some of the effects of sublingual immunotherapy may derive from inflammatory cell recruitment at the site of allergen release . Antitumor activity and molecular effects of the novel heat shock protein 90 inhibitor , IPI-504 , in pancreatic cancer . Targeting Hsp90 is an attractive strategy for anticancer therapy because the diversity and relevance of biological processes are regulated by these proteins in most cancers . However , the role and mode of action of Hsp90 inhibitors in pancreatic cancer has not been studied . This study aimed to assess the antitumor activity of the Hsp90 inhibitor , IPI-504 , in pancreatic cancer and to determine the biological effects of the agent . In vitro , we show that pharmacologic inhibition of Hsp90 by IPI-504 exerts antiproliferative effects in a panel of pancreatic cancer cells in a dose- and time-dependent manner . In pancreatic cancer xenografts obtained directly from patients with pancreas cancer , the agent resulted in a marked suppression of tumor growth . Although known Hsp90 client proteins were significantly modulated in IPI-504-treated cell line , no consistent alteration of these proteins was observed in vivo other than induction of Hsp70 expression in the treated xenografted tumors . Using a proteomic profiling analysis with isotope tags for relative and absolute quantitation labeling technique , we have identified 20 down-regulated proteins and 42 up-regulated proteins on IPI-504 treatment.tumor growth Identical changes were observed in the expression of the genes coding for these proteins in a subset of proteins including P0DMV9 , P17931 , P62158 , Q96KN1 , P14324 , Q8NBJ4 , P69905 , P16403 , HLA-B , and P29966 . The majority of these proteins belong to the functional class of intracellular signal transduction , immune response , cell growth and maintenance , transport , and metabolism . In summary , we show that IPI-504 has potent antitumor activity in pancreatic cancer and identify potential pharmacologic targets using a proteomics and gene expression profiling . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity .

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interacts_with DB08870?

multiple_choice

[ Establishment of hematological malignancy model in ex vivo cell culture ] . The aim of this study was to develop an ex vivo cell culture system for establishing the hematological malignancy model . Mouse bone marrow cells were transfected with GFP-expressed retroviral vectors encoding various leukemia/lymphoma-associated fusion proteins ( P41212 - P09619 , Rabaptin5- P09619 , p210BCR- P00519 , Q01196 - Q06455 , P06748 - Q9UM73 ) . After transfection , the cells were cultured in IMDM containing 10 % FCS without growth factors , or with one of the following growth factor combinations : ( 1 ) murine c-kit ligand ( KL ) plus human flt3 ligand ( FL ) ; ( 2 ) P08700 , thrombopoietin , DB00099 , and hyper- P05231 ( 3/T/G/ Q9NP08 ) ; ( 3 ) KL/FL plus 3/T/G/ Q9NP08 . The flow cytometry was used to detect the ability of combinations of growth factors to complement the oncogene fusion protein to support self-renewal of the transfected cells . The results showed that the transfected cells could be amplified sustainably in the logarithmic growth way . The indicated combination of P21583 ( KL ) with flt-3 ligand ( FL ) supported the self-renewal of the marrow cells transfected with vectors encoding P41212 - P09619 , Rabaptin5- P09619 , Q01196 - Q06455 and P06748 - Q9UM73 , in addition to KL/FL , the self-renewal of Q92817 P11274 - P00519 transfected-marrow cells also required P08700 . The morphology of cells emerged from culture can be the predictor of the corresponding oncogene-associated malignancy . It is concluded that this study establishes a culture system ex vivo which provides a generalized method for studying hematological malignancies , and may facilitate the screening for therapeutic agents . DB08870 : its role in the treatment of anaplastic large cell and Hodgkin 's lymphoma . DB08870 is being developed in a joint collaboration between Seattle Genetics and Millennium : The Takeda Oncology Company . In August 2011 , it was approved by the FDA for the treatment of patients with Hodgkin 's lymphoma ( HL ) and anaplastic large cell lymphoma ( ALCL ) . DB08870 is an antibody-drug conjugate that specifically targets the P01375 receptor superfamily member 8 ( P28908 ) antigen on the surface of cancer cells to induce cell death . DB08870 has shown efficacy in inducing apoptosis in HL and ALCL cell lines that express P28908 and reducing tumor size in preclinical models . DB08870 is under clinical evaluation for the treatment of relapsed or refractory HL and ALCL in both adults and children . It is being investigated for use as a combination agent with pre-existing frontline chemotherapies and as a stand-alone salvage therapy for use prior to autologous stem cell transplant . Treatment with brentuximab vedotin is generally well tolerated although it is associated with grade 1-2 adverse reactions such as neutropenia and there have been reports of grade 3-4 serious adverse events . In particular its use with chemotherapy regimens that include bleomycin is contraindicated because of adverse pulmonary effects . Anti-inflammatory effect of parenteral fish oil lipid emulsion on human activated mononuclear leukocytes . BACKGROUND & AIM : To compare the effect of fish oil-based ( FO ) lipid emulsions ( LE ) for parenteral administration with standard LE and a new FO containing LE composed of four different oils on the antigen presentation and inflammatory variables . METHODS : Phytohemagglutinin ( PHA ) activated human mononuclear leukocytes were cultured with different LE - Control : without LE ; SO : soybean oil ; SO/FO : soybean and FO ( 4:1 ) ; Q8IVS2 /SO : medium chain triglycerides and SO ( 1:1 ) ; Q8IVS2 /SO/FO : Q8IVS2 /SO and FO ( 4:1 ) and SMOF : a new LE containing FO . Cytokine production was evaluated by ELISA , the expression of antigen-presenting and co-stimulatory surface molecules were analyzed by flow cytometry and lymphocyte proliferation was assessed by H(3)- DB04485 incorporation , after tetanus toxoid-induced activation . RESULTS : All LE decreased the HLA-DR and increased P10747 and P16410 expression on monocytes/macrophages and lymphocytes surface ( p < 0.05 ) . SO/FO and Q8IVS2 /SO/FO decreased lymphocyte proliferation ( p < 0.05 ) . All LE decreased P60568 production , but this effect was enhanced with Q8IVS2 /SO/FO and SMOF ( p < 0.05 ) . Q8IVS2 /SO/FO decreased P05231 and increased P22301 , whereas SO had the opposite effect ( p < 0.05 ) . CONCLUSION : FO LE inhibited lymphocyte proliferation and had an anti-inflammatory effect . These effects seem to be enhanced when FO is mixed with Q8IVS2 /SO . SMOF had a neutral impact on lymphocyte proliferation and P05231 and P22301 production . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells . Antibody-drug conjugates ( ADC ) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer . DB08870 represents a first-in-class ADC directed against P28908 (+) malignancies . We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response . In this study , we demonstrate that the dolastatin family of microtubule inhibitors , from which the cytotoxic component of brentuximab vedotin is derived , comprises potent inducers of phenotypic and functional dendritic cell ( DC ) maturation . In addition to the direct cytotoxic effect on tumor cells , dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes . Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells . Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins , the antitumor effect was far less pronounced in immunocompromised mice . We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the P18621 - Q9NZQ7 and P16410 coinhibitory pathways . Ultimately , treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients . Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies , such as brentuximab vedotin , with immune-based therapies . Human lung fibroblasts increase P01730 (+)CD25(+)Foxp3(+) T cells in co-cultured P01730 (+) lymphocytes . Aim of this study was to evaluate functional modifications induced by human lung fibroblasts in co-cultured P01730 (+) T lymphocytes . P01730 (+) T cells , resting or stimulated with ionomycin/PMA for 6h , were co-cultured with fibroblasts isolated from pulmonary biopsies , in contact or separated by a semi-permeable membrane . The expression of CD25 , P16410 , TGF-β , IFNγ , P60568 , P05112 , P22301 and Foxp3 was evaluated by flow cytometric analysis . Fibroblasts induced a significant increment in CD25(+) cells in co-cultured activated P01730 (+) T lymphocytes separated by a membrane . Moreover , fibroblasts treatment with a P35354 inhibitor abrogated the increment in CD25(+) cells whereas exogenous DB00917 restored it . The CD25(+) subpopulation was characterized by increased presence of Fox- P09131 , P16410 , P22301 and TGF-β positive cells while IFN-γ and P60568 positive cells were diminished . Proliferative response of P01730 (+) to the anti CD3/ P10747 -Abs was abrogated in P01730 (+) co-cultured with fibroblasts thus demonstrating a suppressive feature of the expanded CD25(+) subpopulation . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . T cell signaling targets for enhancing regulatory or effector function . To respond to infection , resting or naïve T cells must undergo activation , clonal expansion , and differentiation into specialized functional subsets of effector T cells . However , to prevent excessive or self-destructive immune responses , regulatory T cells ( T(regs) ) are instrumental in suppressing the activation and function of effector cells , including effector T cells . The transcription factor Forkhead box P09131 ( Foxp3 ) regulates the expression of genes involved in the development and function of T(regs) . Foxp3 interacts with other transcription factors and with epigenetic elements such as histone deacetylases ( HDACs ) and histone acetyltransferases . T(reg) suppressive function can be increased by exposure to HDAC inhibitors . The individual contributions of different HDAC family members to T(reg) function and their respective mechanisms of action , however , remain unclear . A study showed that Q9UBN7 , Q9UKV0 , and Sirtuin-1 had distinct effects on Foxp3 expression and function , suggesting that selectively targeting HDACs individually or in combination may enhance T(reg) stability and suppressive function . Another study showed that the receptor programmed death 1 ( P18621 ) , a well-known inhibitor of T cell activation , halted cell cycle progression in effector T cells by inhibiting the transcription of the gene encoding the substrate-recognition component ( Skp2 ) of the ubiquitin ligase P21583 (Skp2) . Together , these findings reveal new signaling targets for enhancing T(reg) or effector T cell function that may be helpful in designing future therapies , either to increase T(reg) suppressive function in transplantation and autoimmune diseases or to block P18621 function , thus increasing the magnitude of antiviral or antitumor immune responses of effector T cells . Augmentation of immune response by chicoric acid through the modulation of P10747 / P16410 and Th1 pathway in chronically stressed mice . This study demonstrates the protective effect of chicoric acid ( CA ) on chronic restraint stress-induced altered T lymphocyte subset distribution and corresponding cytokine secretion patterns in experimental Swiss albino mice . CA has the potential to restore diminished immune response and Th1/Th2 homeostasis in chronically stressed mice as evident by significant increase in lymphocyte proliferation and CD3(+) , P01730 (+) and CD8(+) T cell population . Interestingly , chicoric acid imparted immunostimulation mainly by upregulating the expression of P10747 and P33681 and downregulating P16410 . It exerted stimulatory effect on IL-12 , P01579 and P60568 and suppressed the increased P22301 levels in chronically stressed mice . It also exhibited a significant lowering effect on raised corticosterone levels and reversed the chronic stress-induced hypertrophy of adrenal glands and atrophy of thymus and spleen , thereby showing its normalizing effect on Q9Y251 axis . Our results reveal that CA has the potential to reverse the impact of chronic restraint stress on immune status by normalizing corticosterone levels and augmenting Th1 cytokine profile along with the co-stimulatory molecules particularly P10747 / P16410 pathway that plays a very important role in generation of an effective immune response in immune compromised situations . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . DB00099 -induced stem cell mobilization in chronic myeloid leukaemia patients during imatinib therapy : safety , feasibility and evidence for an efficient in vivo purging . Therapy with imatinib mesylate is limited by cellular resistance in chronic myeloid leukaemia ( CML ) . Further , the limited availability of matching stem cell donors or an unfavourable risk profile for allogeneic stem cell transplantation ( P09683 ) reduces the number of therapeutic options in a number of patients . To assess the possibility of stem cell mobilization ( DB00919 ) during imatinib therapy we performed granulocyte colony-stimulating factor (filgrastim)-induced DB00919 and subsequent aphaeresis in 15 chronic phase and three accelerated phase CML patients . Aphaeresis was successful in 13 patients ( 72 % ) ( > or =2.0 x 10(6) P28906 + cells/kg body weight ) and five ( 28 % ) harvests could be obtained , which were negative for P11274 / P00519 mRNA as assessed by nested-reverse transcription polymerase chain reaction ( RT-PCR ) . All harvests , except one , were negative after first round RT-PCR , implicating a low level of CML cell contamination . There was no significant change in peripheral P11274 / P00519 transcript load after DB00919 as assessed by quantitative real-time RT-PCR . Fifteen patients remained stable in complete cytogenetic remission ( CCR ) during a median observation period of 9.3 months . One patient achieved a molecular remission shortly after DB00919 . Another patient who exhibited rising P11274 / P00519 mRNA levels before DB00919 achieved CCR after autologous P09683 with the generated harvest . One patient with a Philadelphia chromosome-negative , P11274 / P00519 -positive CML showed a cytogenetic relapse 6 months after DB00919 . We conclude that filgrastim-induced P28906 + cell aphaeresis under simultaneous imatinib medication is safe and feasible in CML patients . Additionally , we found evidence that this procedure could generate stem cell harvests that exhibit non-detectable levels of P11274 / P00519 mRNA . In vivo cytokine responses to interleukin-2 immunotherapy after autologous stem cell transplantation in children with solid tumors . The potent immunostimulatory cytokine interleukin-2 ( P60568 ) has been extensively investigated for its potential to induce anti-tumor immunity in a number of tumor models . Only recently the complex interplay of mutually suppressive or supportive cytokines of the P60568 -induced network of cytokines has been better characterized . The aim of this study was to assess which of these in vitro findings are reproducible in vivo in recipients of stem cell transplants ( P09683 ) , since in these patients long- lasting impairments in cytokine inducibility have been described . We have therefore studied the kinetics of putative modulators and mediators of P60568 -induced immune activation , namely IL-1beta , P05112 , P05113 , P22301 , IL-12 , soluble P48023 ( sFasL ) , and GM- P04141 during P60568 therapy . All patients were children or adolescents suffering from solid tumors with poor prognosis who received three 5-day courses of high-dose intravenous P60568 as an adjuvant to their radio-chemotherapy and autologous P09683 . While IL-1beta , P05112 and IL-12 were not , and sFasL was only mildly affected by the P60568 therapy , we observed a consistent and early rise of P22301 , P05113 , and GM- P04141 . These increases were rapidly reversible after discontinuation of P60568 therapy . The inducibility of P22301 , P05113 and GM- P04141 was more pronounced with increasing time from the P09683 , and in the third cycle reached an order of magnitude as in high-dose P60568 patients without P09683 . Together with the abundant in vitro data , these findings may help devise a combination immunotherapy permitting stronger anti-tumor effects , but lesser adverse effects . Altered placental development in pregnancies resulting in sudden infant death syndrome ( P22304 ) . BACKGROUND : Sudden infant death syndrome ( P22304 ) is postulated to be a developmental disorder originating during fetal life in utero . Knowledge regarding the intrauterine environment in which P22304 infants develop is , however , inadequate and how the placenta develops prior to a P22304 event has not been studied . AIM : To investigate the morphological development of the placenta obtained from full-term infants who subsequently succumbed to P22304 . STUDY DESIGN : To estimate the percentage and total volumes of the chorionic villi and villous trophoblast membrane using stereological techniques . SUBJECTS : Placentas were obtained retrospectively from normal birthweight ( P22304 -NBW n=18 ) and small-for-gestational age ( P22304 -SGA , n=14 ) infants who had succumbed to P22304 , and compared to either control ( n=8 ) or SGA placentas ( n=7 ) , respectively . RESULTS : P22304 -NBW placentas displayed evidence of augmented villous growth shown by significantly greater volumes of placental chorionic villi ( gas-exchanging ( GE ) villi ) in comparison to controls ; this was not observed for P22304 -SGA placentas . However , both P22304 -NBW and P22304 -SGA placentas displayed significantly greater volumes of the cytotrophoblast ( CT ) ( P22304 -NBW only ) , syncytiotrophoblast ( P22304 -SGA only ) and syncytial knots ( P09683 -K ) and those displaying apoptotic syncytial nuclei ( AP P09683 -K ) . In contrast , SGA placentas displayed significantly reduced volumes of chorionic villi , GE villi and the villous trophoblast indicating a P22304 -specific effect associated with augmented placental growth . CONCLUSIONS : Our findings provide initial evidence that placental abnormality , although not necessarily causative , may precede a subset of P22304 cases supporting the hypothesis that the origins of P22304 begin during fetal life in utero . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . Aerosol vaccination with AERAS-402 elicits robust cellular immune responses in the lungs of rhesus macaques but fails to protect against high-dose Mycobacterium tuberculosis challenge . Development of a vaccine against pulmonary tuberculosis may require immunization strategies that induce a high frequency of Ag-specific P01730 and CD8 T cells in the lung . The nonhuman primate model is essential for testing such approaches because it has predictive value for how vaccines elicit responses in humans . In this study , we used an aerosol vaccination strategy to administer AERAS-402 , a replication-defective recombinant adenovirus ( rAd ) type 35 expressing Mycobacterium tuberculosis Ags Ag85A , Ag85B , and TB10.4 , in bacillus Calmette-Guérin ( BCG ) -primed or unprimed rhesus macaques . Immunization with BCG generated low purified protein derivative-specific P01730 T cell responses in blood and bronchoalveolar lavage . In contrast , aerosolized AERAS-402 alone or following BCG induced potent and stable Ag85A/b-specific P01730 and CD8 effector T cells in bronchoalveolar lavage that largely produced IFN-γ , as well as P01375 and P60568 . Such responses induced by BCG , AERAS-402 , or both failed to confer overall protection following challenge with 275 CFUs M. tuberculosis Erdman , although vaccine-induced responses associated with reduced pathology were observed in some animals . Anamnestic T cell responses to Ag85A/b were not detected in blood of immunized animals after challenge . Overall , our data suggest that a high M. tuberculosis challenge dose may be a critical factor in limiting vaccine efficacy in this model . However , the ability of aerosol rAd immunization to generate potent cellular immunity in the lung suggests that using different or more immunogens , alternative rAd serotypes with enhanced immunogenicity , and a physiological challenge dose may achieve protection against M. tuberculosis . The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice . Electrolyzed reduced water ( ERW ) is widely used for drinking by people in Asia . The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae . In the non-infected groups , interleukin ( IL ) -4 ( p < 0.001 ) , P05113 , P22301 , IL-1beta , tumor necrosis factor ( P01375 ) -alpha and immunoglobulin ( Ig ) A expression of the group fed ERW ( ERW group ) increased in small intestine compared with the normal control group . In the case of infected groups , the group fed ERW ( ERW+E. hortense group ) showed the result that P05112 , P05113 , P22301 and Ig A expression increased , but IL-1beta and P01375 ( p < 0.001 ) decreased , and the number of goblet cells ( p < 0.001 ) and helix pomatia agglutinin ( Q9Y251 ) positive cells increased compared with the group without feeding ERW . However , adult worm recovery rate was markedly increased ( p < 0.05 ) . On the other hand , the expression of all the cytokines except P22301 in spleen was mildly increased but not significant statistically , and there was no significant difference in the numerical changes of white blood cell ( WBC ) . These results indicate that feeding ERW may have influence on the local immune response ( Th-1 type cytokines such as IL-1beta , P01375 ) in the small intestine but not on the systemic immune response . Results of a pivotal phase II study of brentuximab vedotin for patients with relapsed or refractory Hodgkin 's lymphoma . PURPOSE : DB08870 is an antibody-drug conjugate ( ADC ) that selectively delivers monomethyl auristatin E , an antimicrotubule agent , into P28908 -expressing cells . In phase I studies , brentuximab vedotin demonstrated significant activity with a favorable safety profile in patients with relapsed or refractory P28908 -positive lymphomas . PATIENTS AND METHODS : In this multinational , open-label , phase II study , the efficacy and safety of brentuximab vedotin were evaluated in patients with relapsed or refractory Hodgkin 's lymphoma ( HL ) after autologous stem-cell transplantation ( auto- P09683 ) . Patients had histologically documented P28908 -positive HL by central pathology review . A total of 102 patients were treated with brentuximab vedotin 1.8 mg/kg by intravenous infusion every 3 weeks . In the absence of disease progression or prohibitive toxicity , patients received a maximum of 16 cycles . The primary end point was the overall objective response rate ( ORR ) determined by an independent radiology review facility . RESULTS : The ORR was 75 % with complete remission ( CR ) in 34 % of patients . The median progression-free survival time for all patients was 5.6 months , and the median duration of response for those in CR was 20.5 months . After a median observation time of more than 1.5 years , 31 patients were alive and free of documented progressive disease . The most common treatment-related adverse events were peripheral sensory neuropathy , nausea , fatigue , neutropenia , and diarrhea . CONCLUSION : The ADC brentuximab vedotin was associated with manageable toxicity and induced objective responses in 75 % of patients with relapsed or refractory HL after auto- P09683 . Durable CRs approaching 2 years were observed , supporting study in earlier lines of therapy . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 ) -induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 -Px , MDA , NO and P35228 , and the activities of serum ALT and Q9NRA2 caused by DB00316 . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 ) , pro-inflammatory mediators ( IL-1β , P05112 , P05231 , P01375 -α , P35228 , Bax , HMGB-1 and P35354 ) , pro-inflammatory transcription factors ( NF-κB and AP-1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase-3 , caspase-9 , p-JNK , p-p38 and p- P29323 ) , and increased the protein expressions of Bcl-2 and Bcl-xL . Moreover , the gene expression of P22301 , and the proteins including LC3 , Q14457 and Atg5 induced by DB00316 were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 -induced liver damage in the future . Proteomics and phosphoproteomics provide insights into the mechanism of action of a novel pyrazolo[3,4-d]pyrimidine Src inhibitor in human osteosarcoma . Osteosarcoma ( OS ) is a highly malignant bone tumour , affecting mainly children and young adults between 10 and 20 years of age . It represents the most frequent primitive malignant tumour of the skeletal system and is characterized by an extremely aggressive clinical course , with rapid development of lung metastases . In the last few years , targeting Src in the treatment of OS has become one of the major challenges in the development of new drugs , since an elevated Src kinase activity has been associated with the development and the maintenance of the OS malignant phenotype . Recently , SI-83 , a novel pyrazolo[3,4-d]pyrimidine derivate Src inhibitor , was selected as a promising OS therapeutic drug because of its elevated anti-tumour effects toward human OS . In the present study , gel-based proteomics and phosphoproteomics revealed significant changes in proteins involved in many cancer related processes . We got insight into SI-83 proapoptotic and antiproliferative properties ( overrepresentation of P42261 , P11021 , and P27797 and underrepresentation of P06748 , Q15293 , and P07237 ) . Nevertheless , the most significant findings of our work are the SI-83 induced dephosphorylation of Q9BPX5 , a subunit of the actin related Arp2/3 complex , and the decrease of other cytoskeleton proteins . These data , together with a dramatic impairment of SaOS-2 cell migration and adhesion , suggest that SI-83 may have antimetastatic features that enhance its use as a potent OS chemotherapeutic drug . Niacin reduces plasma P11597 levels by diminishing liver macrophage content in P11597 transgenic mice . The anti-dyslipidemic drug niacin has recently been shown to reduce the hepatic expression and plasma levels of P11597 . Since liver macrophages contribute to hepatic P11597 expression , we investigated the role of macrophages in the P11597 -lowering effect of niacin in mice . In vitro studies showed that niacin does not directly attenuate P11597 expression in macrophages . Treatment of normolipidemic human P11597 transgenic mice , fed a Western-type diet with niacin for 4 weeks , significantly reduced the hepatic cholesterol concentration ( -20 % ) , hepatic P11597 gene expression ( -20 % ) , and plasma P11597 mass ( -30 % ) . Concomitantly , niacin decreased the hepatic expression of P34810 ( -44 % ) and P45844 ( -32 % ) , both of which are specific markers for the hepatic macrophage content . The decrease in hepatic P11597 expression was significantly correlated with the reduction of hepatic macrophage markers . Furthermore , niacin attenuated atherogenic diet-induced inflammation in liver , as evident from decreased expression of P01375 ( -43 % ) . Niacin similarly decreased the macrophage markers and absolute macrophage content in hyperlipidemic P02649 *3-Leiden. P11597 transgenic mice on a Western-type diet . In conclusion , niacin decreases hepatic P11597 expression and plasma P11597 mass by attenuating liver inflammation and macrophage content in response to its primary lipid-lowering effect , rather than by attenuating the macrophage P11597 expression level . A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death . BACKGROUND : Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine . We present a family in which a common polymorphism ( single nucleotide polymorphism ) inherited from the father , combined with a stop codon mutation inherited from the mother ( both asymptomatic ) , led to 2 cases of sudden infant death . METHODS AND RESULTS : P51787 , Q12809 , Q14524 , P15382 , Q9Y6J6 , CACNA1c , CACNB2b , and P63252 genes were amplified and analyzed by direct sequencing . Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary ( CHO- P04264 ) and COS-1 cells . An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis . The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc ( 560 ms ) . The mother was asymptomatic but displayed a prolonged QTc . Genetic screening of the mother revealed a heterozygous nonsense mutation ( P926AfsX14 ) in Q12809 , predicting a stop codon . The father was asymptomatic with a normal QTc but had a heterozygous polymorphism ( K897T ) in Q12809 . The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14 . Heterologous coexpression of K897T and P926AfsX14 led to loss of function of Q12809 current much greater than expression of K897T or P926AfsX14 alone . CONCLUSIONS : Our data suggest that a common polymorphism ( K897T ) can markedly accentuate the loss of function of mildly defective Q12809 channels , leading to long-QT syndrome-mediated arrhythmias and sudden infant death . Protein disulfide isomerases are antibody targets during immune-mediated tumor destruction . The identification of cancer antigens that contribute to transformation and are linked with immune-mediated tumor destruction is an important goal for immunotherapy . Toward this end , we screened a murine renal cell carcinoma cDNA expression library with sera from mice vaccinated with irradiated tumor cells engineered to secrete granulocyte macrophage colony-stimulating factor ( GM- P04141 ) . Multiple nonmutated , overexpressed proteins that function in tumor cell migration , protein/nucleic acid homeostasis , metabolism , and stress responses were detected . Among these , the most frequently recognized clone was protein disulfide isomerase ( P07237 ) . High titer antibodies to human P07237 were similarly induced in an acute myeloid leukemia patient who achieved a complete response after vaccination with irradiated , autologous GM- P04141 -secreting tumor cells in the setting of nonmyeloablative allogeneic bone marrow transplantation . Moreover , Q15084 , a closely related disulfide isomerase involved in major histocompatibility complex ( MHC ) class I chain-related protein A ( Q29983 ) shedding , also evoked potent humoral reactions in diverse solid and hematologic malignancy patients who responded to GM- P04141 -secreting tumor cell vaccines or antibody blockade of cytotoxic T lymphocyte-associated antigen 4 ( P16410 ) . Together , these findings reveal the unexpected immunogenicity of PDIs and raise the possibility that these gene products might serve as targets for therapeutic monoclonal antibodies . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Kinetics of Th1/Th2 cytokines and lymphocyte subsets to predict chronic GVHD after allo- P09683 : results of a prospective study . The role of different cytokines and cells of immune system in the pathogenesis of chronic GVHD ( cGVHD ) is still controversial . Earlier studies , which were either retrospective or analysed one or a few factors , did not show unequivocal results . We prospectively evaluated cytokine levels and lymphocyte subsets in 30 patients who underwent Allo- P09683 to investigate their possible correlation with cGVHD . Levels of P05112 , P05231 , P22301 , P01579 , tumour necrosis factor-alpha ( P01375 ) and its soluble receptors were assessed by ELISA in 30 patients at different times after P09683 . Lymphocyte subsets were evaluated by flow cytometry in peripheral blood at the same times as cytokines . A multivariate analysis was performed using principal component analysis and multi-factor Q9UNW9 ( analysis of variance ) . Eighteen patients developed cGVHD at a median time of 6 months ( range , 5-9 ) after P09683 . In multivariate analysis , we observed a correlation between cGVHD and clusters of cytokines and lymphocyte subsets from the third to the sixth month after P09683 . These clusters changed their composition over time , but they constantly included natural killer ( NK ) and P16410 + T cells as negative predictors of cGVHD . P01375 prevailed among other cytokines before the onset of cGVHD . This prevalence could be related partly to the defect of immunoregulatory cells . 3,4-Dimethoxyphenyl bis-benzimidazole derivative , mitigates radiation-induced DNA damage . Radiation-induced DNA damage initiates a series of overlapping responses that include DNA damage recognition and repair , induction of cell cycle checkpoints , senescence and/or apoptosis . This study assessed the DNA damage response and whole genome expression profile in two mammalian cell lines ( P29320 and U87 ) in response to ( 5-{4-methylpiperazin-1-yl}-2-[2'-(3,4-dimethoxyphenyl)-5'-benzimidazolyl] benzimidazole ) P28067 and ionizing radiation . P28067 has been shown to act as a potent radiation protector , yielding significant levels of protection , i.e. , 20.9 % in P29320 cells and 21.2 % in U87 cells . Our findings revealed treatment with P28067 significantly reduced γ- P16104 , Q12888 and Rad51 foci formation after irradiation . Q96HU1 kinase , WNT signaling and p53 pathways were found to be activated in P28067 -treated cells . In addition , the DNA damage response genes , HSP70 , P10809 , Q06830 , Q9BXM0 , P27797 , P06748 , P0CG48 , and Q01105 showed differential regulation in P28067 , P28067 + radiation and radiation-treated cells . The data suggest that P28067 -influenced repertoire of repair proteins , which are an indispensable part of the cell , interplay with each other to reduce DNA damage and maintain the genomic integrity of the cell . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Q9UM73 ( Q9UM73 ) -induced malignancies : novel mechanisms of cell transformation and potential therapeutic approaches . Among the many oncogenic variants of the anaplastic lymphoma kinase ( Q9UM73 ) , nucleophosmin 1 ( P06748 ) / Q9UM73 fusion protein expressed in the subset of T-cell lymphoma ( Q9UM73 (+) Q9H4E5 ) is currently the best characterized . P06748 / Q9UM73 activates several signal transduction pathways , including PI3K/AKT , MEK/ P29323 , mTORC1 , P40763 , and STAT5b . In turn , the pathways modulate expression and function of many genes and proteins involved in the key cellular functions such as proliferation , growth , survival , metabolism , and angiogenesis . Recent data indicate that P06748 / Q9UM73 also promotes immune evasion of the Q9UM73 (+) Q9H4E5 by inducing through P40763 activation the expression of immunosuppressive cytokines interleukin-10 ( P22301 ) and transforming growth factor-beta ( TGFss ) and cell surface protein Q9NZQ7 ( Q9NZQ7 , Q9NZQ7 ) . In addition , P06748 / Q9UM73 protects its own expression by mediating via P40763 and at least one member of the DNA methyltransferase family P26358 epigenetic silencing of the Q15466 -1 and STAT5a genes . In Q9UM73 + Q9H4E5 cells , Q15466 -1 and STAT5a proteins act as potent tumor suppressors by promoting degradation of the P06748 / Q9UM73 protein and inhibiting expression of the P06748 / Q9UM73 gene , respectively . These findings provide further rationale to therapeutically target Q9UM73 and its effector proteins , foremost P40763 . They also suggest that immunotherapeutic approaches to Q9UM73 (+) Q9H4E5 and , possibly , other Q9UM73 -driven malignancies may require inhibition of Q9UM73 and P40763 to achieve the optimal clinical efficacy . DB08870 followed by allogeneic transplantation as salvage regimen in patients with relapsed and/or refractory Hodgkin 's lymphoma . Patients with relapsed or refractory Hodgkin lymphoma ( RR-HL ) have poor outcomes . DB08870 ( BV ) , an antibody-drug conjugate comprising an anti- P28908 antibody conjugated to the potent anti-microtubule agent , monomethyl auristatin E , induces high tumour responses with moderate adverse effects . In a retrospective study , we describe objective response rates and subsequent allogeneic stem cell transplantation ( allo- P09683 ) in patients with RR-HL treated by BV in a named patient program in two French institutions . Twenty-four adult patients with histologically proven P28908 (+) RR-HL treated with BV were included from July 2009 to November 2012 . Response to BV treatment was evaluated after four cycles . Eleven patients were in complete response ( 45.8 % ) , while five patients were in partial response ( 20.8 % ) , with an overall response rate of 66.6 % . Eight patients failed to respond to BV ( 33.3 % ) . All of the responding patients could receive consolidation treatment after BV : three patients underwent autologous stem cell transplantation ( auto- P09683 ) , three patients received a tandem auto- P09683 /allo- P09683 , nine patients received allo- P09683 and one patient was treated with donor lymphocyte infusion . We found no treatment-related mortality at day 100 among the 12 patients who underwent BV following by allogeneic transplantation . With a median follow-up of 20 months ( range 10.5-43.2 ) , none of them relapsed or died . BV followed by allo- P09683 represents an effective salvage regimen in patients with RR-HL . Hepatic stellate cells undermine the allostimulatory function of liver myeloid dendritic cells via P40763 -dependent induction of P14902 . Hepatic stellate cells ( HSCs ) are critical for hepatic wound repair and tissue remodeling . They also produce cytokines and chemokines that may contribute to the maintenance of hepatic immune homeostasis and the inherent tolerogenicity of the liver . The functional relationship between HSCs and the professional migratory APCs in the liver , that is , dendritic cells ( DCs ) , has not been evaluated . In this article , we report that murine liver DCs colocalize with HSCs in vivo under normal , steady-state conditions , and cluster with HSCs in vitro . In vitro , HSCs secrete high levels of DC chemoattractants , such as MΙP-1α and P13500 , as well as cytokines that modulate DC activation , including P01375 -α , P05231 , and IL-1β . Culture of HSCs with conventional liver myeloid ( m ) DCs resulted in increased P05231 and P22301 secretion compared with that of either cell population alone . Coculture also resulted in enhanced expression of costimulatory ( P33681 , P42081 ) and coinhibitory ( Q9NZQ7 ) molecules on mDCs . P19526 -induced mDC maturation required cell-cell contact and could be blocked , in part , by neutralizing MΙP-1α or P13500 . P19526 -induced mDC maturation was dependent on activation of P40763 in mDCs and , in part , on P19526 -secreted P05231 . Despite upregulation of costimulatory molecules , mDCs conditioned by HSCs demonstrated impaired ability to induce allogeneic T cell proliferation , which was independent of Q9NZQ7 , but dependent upon P19526 -induced P40763 activation and subsequent upregulation of P14902 . In conclusion , by promoting P14902 expression , HSCs may act as potent regulators of liver mDCs and function to maintain hepatic homeostasis and tolerogenicity . Proteomic , cellular , and network analyses reveal new P51452 interactions with nucleolar proteins in HeLa cells . P51452 ( or Vaccinia virus phosphatase P28562 -related ; P51452 ) is a small dual-specificity phosphatase known to dephosphorylate c-Jun N-terminal kinases and extracellular signal-regulated kinases . In human cervical cancer cells , P51452 is overexpressed , localizes preferentially to the nucleus , and plays a key role in cellular proliferation and senescence triggering . Other P51452 functions are still unknown , as illustrated by recent and unpublished results from our group showing that this enzyme mediates DNA damage response or repair processes . In this study , we sought to identify new interactions between P51452 and proteins directly or indirectly involved in or correlated with its biological roles in HeLa cells exposed to gamma or UV radiation . By using Q86UG4 -DUSP as bait , we pulled down interacting proteins and identified them by LC-MS/MS . Of the 46 proteins obtained , six hits were extensively validated by immune techniques ; the proteins P06748 , HnRNP C1/ P06681 , and P19338 were the most promising targets found to directly interact with P51452 . We then analyzed the P51452 interactomes using physical protein-protein interaction networks using our hits as the seed list . The validated hits as well as unvalidated hits fluctuated on the P51452 interactomes of HeLa cells , independent of the time post radiation , which confirmed our proteomic and experimental data and clearly showed the proximity of P51452 to proteins involved in processes intimately related to DNA repair and senescence , such as P12956 and Tert , via interactions with nucleolar proteins , which were identified in this study , that regulate DNA/RNA structure and functions . Association between high interleukin-6 levels and adverse outcome after autologous haemopoietic stem cell transplantation . We studied interleukin-6 ( P05231 ) levels on the day of transplantation in 31 patients undergoing autologous haemopoietic stem cell transplantation ( P09683 ) ( either peripheral blood stem cell transplantation ( PBSCT ) or bone marrow transplantation ( BMT ) ) for neoplastic diseases to determine if there was a relationship between P05231 level and rate of haemopoietic recovery , length of stay in hospital , and survival . There was no apparent delay in post-transplant recovery associated with elevated P05231 levels . However , increased values of P05231 tended to be associated with an increased length of stay in hospital ( P = 0.083 ) . There was a highly significant adverse association between higher P05231 levels and survival following transplantation ( P = 0.0001 ) . This association remained significant ( P = 0.013 ) in the uniform subgroup of patients with malignant lymphoma with chemosensitive disease who had undergone BMT ( that is , excluding patients who had undergone PBSCT ) ( n = 13 ) . Knowledge of P05231 levels on the day of transplant has the potential to provide valuable prognostic information in patients undergoing autologous haemopoietic P09683 . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Malignant transformation of P01730 + T lymphocytes mediated by oncogenic kinase P06748 / Q9UM73 recapitulates P60568 -induced cell signaling and gene expression reprogramming . Q9UM73 ( Q9UM73 ) , physiologically expressed only by nervous system cells , displays a remarkable capacity to transform P01730 (+) T lymphocytes and other types of nonneural cells . In this study , we report that activity of nucleophosmin ( P06748 ) / Q9UM73 chimeric protein , the dominant form of Q9UM73 expressed in T cell lymphomas ( TCLs ) , closely resembles cell activation induced by P60568 , the key cytokine supporting growth and survival of normal P01730 (+) T lymphocytes . Direct comparison of gene expression by Q9UM73 (+) Q9H4E5 cells treated with an Q9UM73 inhibitor and P60568 -dependent Q9UM73 (-) Q9H4E5 cells stimulated with the cytokine revealed a very similar , albeit inverse , gene-regulation pattern . Depending on the analysis method , up to 67 % of the affected genes were modulated in common by P06748 / Q9UM73 and P60568 . Based on the gene expression patterns , Jak/ P35610 - and P60568 -signaling pathways topped the list of pathways identified as affected by both P60568 and P06748 / Q9UM73 . The expression dependence on P06748 / Q9UM73 and P60568 of the five selected genes-CD25 ( IL-2Rα ) , Egr-1 , Fosl-1 , O14543 , and Irf-4-was confirmed at the protein level . In both Q9UM73 (+) Q9H4E5 and P60568 -stimulated Q9UM73 (-) Q9H4E5 cells , CD25 , O14543 , and Irf-4 genes were activated predominantly by the P42229 and P40763 transcription factors , whereas transcription of Egr-1 and Fosl-1 was induced by the MEK- P29323 pathway . Finally , we found that Egr-1 , a protein not associated previously with either P60568 or Q9UM73 , contributes to the cell proliferation . These findings indicate that P06748 / Q9UM73 transforms the target P01730 (+) T lymphocytes , at least in part , by using the pre-existing , P60568 -dependent signaling pathways . DB08870 in anaplastic large cell lymphoma . INTRODUCTION : DB08870 , a novel anti- P28908 antibody-drug conjugate , delivers a cytotoxic agent into P28908 (+) cells . P28908 expression is characteristic of anaplastic large cell lymphoma ( ALCL ) and Hodgkin lymphoma ( HL ) . AREAS COVERED : We reviewed data on brentuximab vedotin , focusing on ALCL and discuss pharmacology , clinical trials leading to approval and future research directions . Systemic ALCL , 3 % of adult Q9NZ71 , is characterized by large anaplastic P28908 (+) cells . The fusion protein P06748 - Q9UM73 , when present in systemic ALCL , confers better prognosis , although even Q9UM73 - patients with IPI score ≥ 3 are high-risk . For patients with systemic ALCL , 25 - 45 % relapse after frontline therapy , and > 50 % of patients will relapse following high-dose chemotherapy with autologous stem-cell support . There has been no standard therapy for relapsed/refractory systemic ALCL . DB08870 , combines a monoclonal antibody targeted to P28908 with a microtubule disrupting agent and was recently approved for treatment of patients with systemic ALCL that is refractory or relapsed after at least one multiagent chemotherapy regimen . EXPERT OPINION : DB08870 provides targeted therapy to P28908 (+) lymphomas , including ALCL and HL , with high response rates and manageable toxicity , predominantly myelosuppression and peripheral neuropathy . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . The polymorphism IL-1beta T-31C is associated with a longer overall survival in patients with multiple myeloma undergoing auto- P09683 . Proinflammatory cytokines are suspected to play a role in the pathogenesis of multiple myeloma ( MM ) . Therefore , it is possible that inborn genetic variations leading to a modified expression of these cytokines will influence the outcome for these patients . We investigated 348 MM patients undergoing high-dose melphalan treatment followed by Auto- P09683 and examined the influence of single nucleotide polymorphisms ( SNPs ) in genes involved in the inflammatory response . We found that the polymorphism IL-1beta T-31C significantly influenced overall survival ( OS ; P=0.02 ) and that carriers of the variant C-allele had a significantly longer survival than homozygous wild-type allele TT-carriers ( relative risk 0.6 ( 95 % CI=0.5-0.9 ) ; P=0.008 ) . The polymorphisms P05231 G-174C , P22301 C592A , PPARgamma2 Pro(12)Ala , P35354 A-1195G , P35354 T8473C and P19838 ins/del did not influence the OS in this group of patients . Furthermore , homozygous carriers of the variant allele of IL-1beta T-31C were at 1.37-fold ( CI=1.05-1.80 ) increased risk of MM as compared with population-based controls ( P=0.02 ) . Our results indicate that IL-1beta is involved in the pathogenesis of MM . Design and expression of soluble P16410 variable domain as a scaffold for the display of functional polypeptides . We have designed and engineered the human cytotoxic T-lymphocyte associated protein-4 ( P16410 ) variable ( V-like ) domain to produce a human-based protein scaffold for peptide display . First , to test whether the P16410 CDR-like loops were permissive to loop replacement/insertion we substituted either the P51861 or CDR3 loop with somatostatin , a 14-residue intra-disulfide-linked neuropeptide . Upon expression as periplasmic-targeted proteins in Escherichia coli , molecules with superior solubility characteristics to the wild-type V-domain were produced . These mutations in P16410 ablated binding to its natural ligands P33681 and P42081 , whereas binding to a conformation-dependent anti- P16410 monoclonal antibody showed that the V-domain framework remained correctly folded . Secondly , to develop a system for library selection , we displayed both wild-type and mutated P16410 proteins on the surface of fd-bacteriophage as fusions with the geneIII protein . P16410 displayed on phage bound specifically to immobilized P33681 -Ig and P42081 -Ig and in one-step panning enriched 5,000 to 2,600-fold respectively over wild-type phage . Bacteriophage displaying P16410 with somatostatin in CDR3 ( CTLA-4R-Som3 ) specifically bound somatostatin receptors on transfected CHO- P04264 cells pre-incubated with 1 microg/ml tunicamycin to remove receptor glycosylation . Binding was specific , as 1 microM somatostatin successfully competed with CTLA-4R-Som3 . CTLA-4R-Som3 also activated as well as binding preferentially to non-glycosylated receptor subtype Sst4 . The ability to substitute CDR-like loops within P16410 will enable design and construction of more complex libraries of single V-like domain binding molecules . Proteins 1999;36:217-227 . Engagement of CD153 ( P32971 ) by P28908 + T cells inhibits class switch DNA recombination and antibody production in human IgD+ IgM+ B cells . CD153 ( P32971 ) is a member of the P01375 ligand/cytokine family expressed on the surface of human B cells . Upon exposure to P05112 , a critical Ig class switch-inducing cytokine , Ag-activated T cells express P28908 , the CD153 receptor . The observation that dysregulated IgG , IgA , and/or IgE production is often associated with up-regulation of T cell P28908 prompted us to test the hypothesis that engagement of B cell CD153 by T cell P28908 modulates Ig class switching . In this study , we show that IgD+ IgM+ B cells up-regulate CD153 in the presence of CD154 ( P29965 ) , P05112 , and B cell Ag receptor engagement . In these cells , CD153 engagement by an agonistic anti-CD153 mAb or T cell P28908 inhibits S mu --> Sgamma , Smu --> Salpha , and S mu --> Sepsilon class switch DNA recombination ( CSR ) . This inhibition is associated with decreased TNFR-associated factor-2 binding to P25942 , decreased NF-kappaB binding to the P25942 -responsive element of the Cgamma3 promoter , decreased Igamma3-Cgamma3 germline gene transcription , and decreased expression of P12956 , P13010 , DNA protein kinase , switch-associated protein-70 , and Msh2 CSR-associated transcripts . In addition , CD153 engagement inhibits IgG , IgA , and IgE production , and this effect is associated with reduced levels of B lymphocyte maturation protein-1 transcripts , and increased binding of B cell-specific activation protein to the Ig 3' enhancer . These findings suggest that P28908 + T cells modulate CSR as well as IgG , IgA , and IgE production by inducing reverse signaling through B cell CD153 . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Block of Q12809 channels by berberine : mechanisms of voltage- and state-dependence probed with site-directed mutant channels . DB04115 prolongs the duration of cardiac action potentials without affecting resting membrane potential or action potential amplitude . Controversy exists regarding whether berberine exerts this action by preferential block of different components of the delayed rectifying potassium current , I(Kr) and I(Ks) . Here we have studied the effects of berberine on hERG ( I(Kr) ) and P51787 / P15382 ( I(Ks) ) channels expressed in P29320 -293 cells and Xenopus oocytes . In P29320 -293 cells , the IC50 for berberine was 3.1 +/- 0.5 microM on hERG compared with 11 +/- 4 % decreases on P51787 / P15382 channels by 100 microM berberine . Likewise in oocytes , hERG channels were more sensitive to block by berberine ( IC50 = 80 +/- 5 microM ) compared with P51787 / P15382 channels ( approximately 20 % block at 300 microM ) . hERG block was markedly increased by membrane depolarization . Mutation to Ala of Y652 or F656 located on the S6 domain , or V625 located at the base of the pore helix of hERG decreased sensitivity to block by berberine . An inactivation-deficient mutant hERG channel ( G628C/S631C ) was also blocked by berberine . Together these findings indicate that berberine preferentially blocks the open state of hERG channels by interacting with specific residues that were previously reported to be important for binding of more potent antagonists . Interaction of cyclooxygenase-2 variants and smoking in pancreatic cancer : a possible role of nucleophosmin . BACKGROUND & AIMS : Overexpression of cyclooxygenase-2 ( P35354 ) is implicated in cancer development . This study examined the functional relevance of genetic polymorphisms in the P35354 promoter and evaluated their associations with susceptibility to pancreatic cancer . METHODS : Genotypes and haplotypes of P35354 -765G/C , -1195G/A , and -1290A/G were analyzed in 393 pancreatic cancer patients and 786 controls . Odds ratios ( ORs ) and 95 % confidence intervals ( CIs ) were computed by logistic regression.The function of the -765G --> C polymorphism was examined by a set of biochemical assays . RESULTS : The -1195AA or -765GC genotype carriers had a 1.34-fold ( 95 % CI : 1.12-1.60 ) or 1.63-fold ( 95 % CI : 1.25-2.10 ) excess risk for developing pancreatic cancer . These 2 variants showed a cooperative effect in context of haplotype , with the ORs for the A(-1195)-C(-765)- containing haplotypes being significantly greater than those for the G(-1195)-G(-765)-containing haplotypes . The -765C allele and smoking displayed a multiplicative joint effect , with an OR of 3.72 ( 95 % CI : 1.70-8.14 ) for heavy smokers carrying the -765GC genotype . Biochemical assays suggest that the -765G --> C change creates a binding site for nucleophosmin ( P06748 ) and phosphorylated P06748 ( p- P06748 ) , which acts as a transcriptional inhibitor . Cigarette smoke remarkably increased P35354 promoter activity , and this effect was more pronounced for the -765C allele compared with the -765G allele . Cigarette smoke reduced nuclear p- P06748 levels , which was reversely associated with P35354 expression . CONCLUSIONS : Functional P35354 polymorphisms are associated with susceptibility to pancreatic cancer and tobacco smoke specifically increases -765C promoter activity , which might be mediated by p- P06748 . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared .

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Anti-angiogenic effects of Hypericin-photodynamic therapy in combination with DB00482 in the treatment of human nasopharyngeal carcinoma . Photodynamic therapy ( PDT ) is being investigated as an alternative treatment modality in cancer treatment . It has been shown to induce tumor hypoxia and upregulation of cyclooxygenase-2 ( P35354 ) and vascular endothelial growth factor ( P15692 ) . The objective of this study was to improve in vivo tumor growth control of nasopharyngeal carcinoma ( NPC ) , treated at a subcurative dosage by using a combination of Hypericin-PDT and P35354 inhibitor , DB00482 ( CX ) . The effect of an initial CX dose at 6- and 24-h post-PDT was investigated simultaneously . It was observed that hypoxic NPC/CNE2 cells upregulate both P35354 and P15692 A genes in vitro . In vivo studies , down-regulation of P35354 and hypoxia inducible factor-1alpha ( HIF-1alpha ) genes at 24-h post-PDT and bulk tumor ablation at 48-h post-PDT was observed . However , 24-28 days later regrowth was observed . In a combination treatment , 1st CX dose at 6-h post-PDT had the highest tumor control in which tumors were < or=0.52 cm(3) ( 64.29 % , P < 0.05 ) . However , the tumors administered with a initial dose of CX at 24-h post-PDT had no tumor control . Co-suppression of P35354 , HIF-1alpha and P15692 A genes were observed in tumors with tumor control . Mice without tumor control that were subjected to therapy had increased human P15692 in serum compared to Hypericin-PDT mice . Thus , suggesting that the time of initial administration of CX post-PDT is an important factor for effective tumor control . [ Selective inhibitors of type 2 cyclooxygenase : less renal effects than the classical non-steroidal anti-inflammatory agents ] . Prostaglandins play an important role in the regulation of renal hemodynamics and sodium excretion . Thus , the administration of non-steroidal anti-inflammatory drugs ( NSAIDs ) induces a renal vasoconstriction and sodium and potassium retention . In some high risk patients , this may lead to acute renal failure . The anti-inflammatory and renal effects of conventional NSAIDs are mediated by the non-selective inhibition of two cyclo-oxygenases , P23219 and P35354 . Recently , highly selective P35354 inhibitors have been developed such as celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . These drugs were designed to preserve the analgesic and anti-inflammatory properties of NSAIDs while reducing their gastro-intestinal and renal side effects . Selective P35354 inhibitors have indeed less gastro-intestinal side-effects . However , their renal profile is comparable to that of non-selective inhibitors as they induce a decrease in glomerular filtration rate and a sodium and potassium retention . Thus , despite the good gastro-intestinal safety profile of selective P35354 , one should be careful with the use of these agents in high risk patients as they may induce similar renal complications as non-selective NSAIDs . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Inhibition of delayed rectifier potassium channels and induction of arrhythmia : a novel effect of celecoxib and the mechanism underlying it . Selective inhibitors of cyclooxygenase-2 ( P35354 ) , such as rofecoxib ( Vioxx ) , celecoxib ( DB00482 ) , and valdecoxib ( Bextra ) , have been developed for treating arthritis and other musculoskeletal complaints . Selective inhibition of P35354 over P23219 results in preferential decrease in prostacyclin production over thromboxane A2 production , thus leading to less gastric effects than those seen with nonselective P36551 inhibitors such as acetylsalicylic acid ( aspirin ) . Here we show a novel effect of celecoxib via a mechanism that is independent of P35354 inhibition . The drug inhibited the delayed rectifier ( Kv2 ) potassium channels from Drosophila , rats , and humans and led to pronounced arrhythmia in Drosophila heart and arrhythmic beating of rat heart cells in culture . These effects occurred despite the genomic absence of cyclooxygenases in Drosophila and the failure of acetylsalicylic acid , a potent inhibitor of both P23219 and P35354 , to inhibit rat Kv2.1 channels . A genetically null mutant of Drosophila Shab ( Kv2 ) channels reproduced the cardiac effect of celecoxib , and the drug was unable to further enhance the effect of the mutation . These observations reveal an unanticipated effect of celecoxib on Drosophila hearts and on heart cells from rats , implicating the inhibition of Kv2 channels as the mechanism underlying this effect . Genetic tools to tailor cancer prevention by NSAIDs . It was shown that NSAIDs , such as aspirin or DB00482 , are effective cancer preventive agents when taken regularly . However , the long-term use of NSAIDs , the cyclooxygenase ( P36551 ) inhibitors , may have significant adverse effects - primarily on the gastrointestinal ( inhibiting P23219 ) and cardiovascular ( inhibiting P35354 ) systems . Genetic analysis of enzymes ( including P36551 ) involved in the prostaglandin synthesis should reveal and predict a person 's benefits vs. toxicity resulting from the NSAID treatment . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . P35354 inhibition attenuates antibody responses against human papillomavirus-like particles . Vaccination to generate protective humoral immunity against infectious disease is becoming increasingly important due to emerging strains of virus , poorly immunogenic vaccines , and the threat of bioterrorism . We demonstrate that cyclooxygenase-2 ( Cox-2 ) is crucial for optimal Ab responses to a model vaccine , human papillomavirus type 16 virus-like particles ( HPV 16 VLPs ) . Cox-2-deficient mice produce 70 % less IgG , 50 % fewer Ab-secreting cells , and 10-fold less neutralizing Ab to HPV 16 VLP vaccination compared with wild-type mice . The reduction in Ab production by Cox-2(-/-) mice was partially due to a decrease in class switching . SC-58125 , a structural analog of the Cox-2-selective inhibitor DB00482 reduced by approximately 70 % human memory B cell differentiation to HPV 16 VLP IgG-secreting cells . The widespread use of nonsteroidal anti-inflammatory drugs and Cox-2-selective inhibitory drugs may therefore reduce vaccine efficacy , especially when vaccines are poorly immunogenic or the target population is poorly responsive to immunization . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . P35354 inhibitors and DB00482 : safe or suspect ? Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . P35354 promotes early atherosclerotic lesion formation in ApoE-deficient and C57BL/6 mice . Cyclooxygenase ( P36551 ) 2 is expressed in atherosclerotic lesions . We have previously reported that selective inhibition of P35354 reduces early atherosclerosis in P01130 deficient mice . To examine the role of P35354 in atherosclerosis in other mouse models , we studied the effects of selective P35354 inhibition ( by rofecoxib and NS-398 ) and nonselective P36551 inhibition ( by indomethacin ) on early atherosclerotic lesion formation in apolipoprotein E-deficient ( apoE(-/-) ) mice . Selective P35354 and nonselective P36551 inhibition reduced atherosclerosis in female apoE(-/-) mice by 35-38 % and 38-51 % in the proximal and en face aortas , respectively . Next we investigated the role of macrophage P35354 by transplanting P35354 (-/-) fetal liver cells into C57BL/6 mice and challenging the mice with an atherogenic diet . Genetic deletion of P35354 from hematopoietic cells reduced atherosclerosis by 51 % . In addition , LPS activated P35354 (-/-) macrophages had decreased expression of monocyte chemoattractant protein-1 ( P13500 ) and tumor necrosis factor-alpha ( TNFalpha ) . The results demonstrate that selective inhibition of P35354 and elimination of P35354 from macrophages significantly reduces early atherosclerotic lesion formation in apoE-deficient and C57BL/6 mice . These results are compatible with P35354 expression by macrophages having a proatherogenic role , and support the potential of anti-inflammatory therapeutic approaches for atherosclerosis . Compositions for treatment of cancer and inflammation . Celecoxib ( DB00482 , Pfizer , NY , USA ) is a worldwide top branded P35354 -specific inhibitor . It was shown to provide relief of arthritic pain and inflammation and has recently been under investigation for the prevention and treatment of cancer . However , recent studies showed that long term use of high doses of celecoxib is associated with an increased cardiovascular toxicity . We discovered that the addition of curcumin , a natural P35354 inhibitor , to celecoxib synergistically ( up to 1000 % ) augments the growth inhibitory effects of celecoxib in in-vitro and in-vivo models of arthritis and cancer , thus rendering effective action of the drug at up to tenfold lower dose . This may pave the way for a novel strategy to treat arthritis and cancer because its effect [ 1 ] can be achieved in the serum of patients receiving standard anti inflammatory or anti-neoplastic dosages of celecoxib , and [ 2 ] involves a regimen with a very low profile of side effects . Preliminary data suggest that the combination is not limited only to celecoxib and that addition of curcumin to other NSAIDs such as sulindac , synergistically augments neoplastic cell growth inhibition . Based on these finding we received an IRB approval to evaluate celecoxib+curcumin in patients with osteoarthritis , pancreatic cancer and metastatic CRC . We hope to complete these novel human clinical trials , in 12-18 months . Phospholipase A₂ activities in skin physiology and pathology . Skin inflammatory diseases are most commonly treated with corticosteroids , especially topical preparations , benefitting from high potency and unparalleled formulation flexibility . However , these benefits are limited due to side effects , especially under long-term use . Non-steroidal anti-inflammatory drugs ( NSAIDs ) which block the P36551 pathways have been used as safer alternatives to corticosteroids , and much effort and resources have been invested in developing P36551 inhibitors . However , synthetic NSAIDs are less potent than steroids , have limited formulation flexibility and have their own safety issues , thereby yielding unsatisfactory results , with some high-profile drugs ( e.g. , the P35354 inhibitors Vioxx , DB00482 ) being withdrawn from the market due to safety concerns . The potency and safety challenges of NSAIDs are related to inter-eicosanoid dynamics , pertaining to their pro-versus anti-inflammatory action , homeostatic functions and tissue-specific activities . Instead , the upstream control of phospholipase A2 ( P04054 ) enzymatic activity , which hydrolyzes cell membrane phospholipids to initiate the eicosanoid production , has been considered for inhibiting eicosanoid activation while maintaining the intricate balance needed for their homeostatic functions . Yet , PLA(2) inhibitors have hardly been tested for treating skin inflammatory/allergic conditions . In this article we review the involvement of PLA(2)s in skin physiology and pathology , and discuss the prospect of PLA(2) inhibition for the treatment of dermatological diseases . Celecoxib and radiation therapy in non-small-cell lung cancer . Overexpression of cyclooxygenase-2 ( P35354 ) is frequently present in lung cancer and may play a significant role in carcinogenesis , invasion , and metastasis . It has been associated with shortened survival in patients with resected early-stage adenocarcinoma of the lung . P35354 inhibition decreases tumor cell proliferation in vivo and has been shown to enhance tumor radiosensitivity . Additionally , P35354 inhibition may protect normal pulmonary tissue from radiation fibrosis . Clinical studies are under way to assess the potential benefits and risks of P35354 inhibition in the treatment of lung cancer . The rationale for P35354 inhibitors in the treatment of lung cancer will be reviewed . The results of a phase II study assessing the acute toxicity of concurrent celecoxib ( DB00482 ) and thoracic irradiation in patients with non-small-cell lung cancer ( NSCLC ) are reported , and an ongoing Radiation Therapy Oncology Group study using celecoxib and concurrent radiation therapy for NSCLC in patients with intermediate prognostic factors is reviewed . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity . Drug insight : cyclo-oxygenase 2 inhibitors and cardiovascular risk -- where are we now ? Cyclo-oxygenase ( P36551 ) 1 mediates the production of thromboxane A2 in platelets , leading to platelet aggregation and vasoconstriction . Conversely , P35354 catalyzes endothelial prostacyclin synthesis , which effectively counteracts thromboxane A2 , triggering vasodilation and platelet inhibition . Selective P35354 inhibitors decrease prostacyclin production , potentially disrupting homeostasis and creating a prothrombotic state . The VIGOR study findings of increased cardiovascular risk with rofecoxib were subsequently confirmed by large meta-analyses , observational studies and recent APPROVe trial publication . The P25054 trial findings of increased cardiovascular risk with DB00482 ( celecoxib ) conflict with those in the ADAPT trial , the upcoming PreSAP publication , a case-control study by Graham et al. and prior large clinical trials , meta-analyses and observational studies of this drug . Therefore , while an adverse class effect is a possibility for P35354 inhibitors , the published data are inconsistent . Baseline cardiovascular risk in patients might contribute significantly to these findings . In light of the negative Vioxx ( rofecoxib ) publicity , however , P35354 inhibitors might forever remain underinvestigated . The relative selectivity of these compounds for P35354 is extremely variable , casting significant doubt on the class-effect hypothesis . Improved endothelial function has also been reported with celecoxib , leading to endothelium-dependent vasodilation , and associated decreases in P02741 and LDL cholesterol . The addition of meloxicam to low-dose aspirin and heparin has improved clinical outcomes after acute coronary syndromes . These are the first studies suggesting improvement in endothelial function and reduction of inflammation with P35354 inhibition . Thus , more randomized controlled trials are needed to study the relative cardiovascular effects of different P35354 inhibitors , alone and in combination with aspirin . The new P35354 inhibitors : rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) . Celecoxib : a novel treatment for lung cancer . Lung cancer is by far the leading cause of cancer-related deaths . Overall survival is poor and has not improved substantially over the last 50 years . Therefore , it is clear that novel and more effective treatments are needed to improve the outcome of therapy . Recent attention has been drawn to the role of cyclooxygenase ( P36551 ) -2 in the pathogenesis of cancer , and it has been considered as an attractive target for therapeutic and chemopreventive strategies in lung cancer patients . Celecoxib ( DB00482 ) , Pfizer ) , a selective P35354 inhibitor and potent anti-inflammatory agent , has been approved for the treatment of osteoarthritis and rheumatoid arthritis . This orally administered agent is generally well tolerated and has almost no gastrointestinal or renal toxicity . Phase II clinical trials suggest that P35354 inhibition by celecoxib would enhance response to cytotoxic chemotherapy or radiation therapy through interference with cellular proliferation and tumor angiogenic processes , promotion of apoptosis and immune surveillance , or other possible mechanisms . Celecoxib has shown promising antitumor efficacy in lung cancer and a large variety of solid tumors that rely on P35354 -related mechanisms for growth and survival . This article reviews the profile of celecoxib and evidence supporting its role in the therapy of lung cancer . P01308 generates free radicals in human fibroblasts ex vivo by a protein kinase C-dependent mechanism , which is inhibited by pravastatin . P01308 can generate oxygen free radicals . Statins , P04035 inhibitors , exert a powerful antioxidant effect . The present study aimed to clarify the mechanisms through which insulin generates free radicals and to assess whether pravastatin modulates such effects . In cultured skin fibroblasts from human volunteers exposed to high insulin concentration , either in the presence or in the absence of pravastatin , insulin induced translocation of the p47(phox) subunit of NAD(P)H oxidase from the cytosol to the membrane and generation of radical oxygen species through a PKC delta-dependent mechanism . The insulin-induced translocation of p47(phox) was PKC delta dependent and attenuated by pravastatin , but independent of the activation of Akt and Rac1 . P01308 -induced Akt phosphorylation was increased by pravastatin and P27361 /2 phosphorylation attenuated . The present study demonstrates a novel mechanism by which insulin stimulates the generation of free radicals in human fibroblasts , ex vivo . It involves phosphatidylinositol 3-kinase , PKC delta , and p47(phox) translocation and promotes P27361 /2 phosphorylation . DB00175 inhibited radical oxygen species production by inhibiting PKC delta . These observations offer a robust explanation for the positive effects of pravastatin treatment in patients with insulin resistance syndrome . Inflammation induces mitochondrial dysfunction and dopaminergic neurodegeneration in the nigrostriatal system . Evidence suggests that chronic inflammation , mitochondrial dysfunction , and oxidative stress play significant and perhaps synergistic roles in Parkinson 's disease ( PD ) , where the primary pathology is significant loss of the dopaminergic neurons in the substantia nigra . The use of anti-inflammatory drugs for PD treatment has been proposed , and inhibition of cyclo-oxygenase-2 ( P35354 ) or activation of peroxisome proliferator-activated receptor gamma ( P37231 ) yields neuroprotection in MPTP-induced PD . Lipopolysaccharide ( LPS ) induces inflammation-driven dopaminergic neurodegeneration . We tested the hypothesis that celecoxib ( DB00482 , P35354 inhibitor ) or pioglitazone ( Actos , P37231 agonist ) will reduce the LPS-induced inflammatory response , spare mitochondrial bioenergetics , and improve nigral dopaminergic neuronal survival . Rats were treated with vehicle , celecoxib , or pioglitazone and were intrastriatally injected with LPS . Inflammation , mitochondrial dysfunction , oxidative stress , decreased dopamine , and nigral dopaminergic neuronal loss were observed post-LPS . Celecoxib and pioglitazone provided neuroprotective properties by decreasing inflammation and restoring mitochondrial function . Pioglitazone also attenuated oxidative stress and partially restored striatal dopamine as well as demonstrated dopaminergic neuroprotection and reduced nigral microglial activation . In summary , intrastriatal LPS served as a model for inflammation-induced dopaminergic neurodegeneration , anti-inflammatory drugs provided protective properties , and pioglitazone or celecoxib may have therapeutic potential for the treatment of neuro-inflammation and PD . [ Pharma-clinics. The drug of the month. Celecoxib ( DB00482 ) ] . Celecoxib ( DB00482 , Pharmacia ) is a potent and selective inhibitor of the P35354 isoform of cyclooxygenase which is used as nonsteroidal anti-inflammatory drug ( NSAID ) . Its current indications are osteoarthritis and rheumatoid arthritis . The usual recommended daily dosage of celecoxib is 200 mg ( in one or two intakes per day ) , to be increased up to 400 mg ( two intakes per day ) if necessary . Its clinical efficacy seems to be similar to that of other NSAIDs . However , its safety profile , especially gastro-intestinal tolerance and perhaps renal safety , is much better because of the P35354 selectivity . DB00917 produced by lung cancer suppresses immune function through T-regulatory cells and can be blocked by the P35354 inhibitor DB00482 . Plasma levels of vascular endothelial growth factor during and after radiotherapy in combination with celecoxib in patients with advanced head and neck cancer . DB00482 and radiotherapy in advanced head and neck cancer . This phase I dose-escalation study seeks to determine the phase II recommended dose of cyclooxygenase type 2 ( P35354 ) inhibitor in patients with locally advanced squamous cell head and neck ( H & N ) cancer , treated with accelerated radiotherapy . Anti-vasculogenic effect of this treatment on serum vascular endothelial growth factor ( P15692 ) is examined . Patients were irradiated with curative intent ( 72Gy in 6weeks ) . Celecoxib was administered throughout the radiotherapy course . Serum P15692 level were tested during radiotherapy and in follow-up . Tumor specimens were stained to quantify the P35354 expression . Thirty-two patients completed the treatment . The dose of celecoxib was escalated ( 200 , 400 and 800mg bid , then de-escalated to 600mg bid ) . The acute toxicity related to the treatment in the first and second cohort did not reach grade III ; in the third cohort three patients had grade III radiation toxicity and one had celecoxib-related toxicity . In the last fourth cohort the toxicity was acceptable . Significant P15692 level drop ( p=0.011 ) was found between radiation day 1 and post-treatment visit . Significant decrease ( p=0.022 ) of the P15692 level was shown in patients with high P35354 expression in the tumor . Phase II recommended dose of celecoxib combined with accelerated radiotherapy in advanced H & N cancer was 600mg bid . A significant decrease of the post-treatment serum P15692 level compared to the initial level was noticed only in patients with high P35354 expression in tumors . The role of vascular endothelial growth factor and matrix metalloproteinases in canine lymphoma : in vivo and in vitro study . BACKGROUND : Canine lymphoma represents the most frequent haematopoietic cancer and it shares some similarities with human non-Hodgkin lymphoma . Matrix metalloproteinases ( MMPs ) and vascular endothelial growth factor ( P15692 ) play a coordinated role during invasion and proliferation of malignant cells ; however , little is known about their role in canine haematologic malignancies . The aim of this study was to investigate the mRNA and protein expression of P15692 and the most relevant MMPs in canine lymphoma . Lymph node aspirates from 26 B-cell and 21 T-cell lymphomas were collected . The protein expression levels of P14780 , P08253 and P15692 were evaluated by immunocytochemistry , and the mRNA levels of P08253 , P14780 , P50281 , P01033 , P16035 , O95980 , P15692 and P15692 -164 were measured using quantitative RT-PCR . RESULTS : P50281 , P01033 and O95980 mRNA levels were significantly higher in T-cell lymphomas than in B-cell lymphomas . Higher mRNA and protein levels of P14780 and P15692 were observed in T-cell lymphomas than in B-cell lymphomas and healthy control lymph nodes . A positive correlation was found between P14780 and P15692 in T-cell lymphomas . Moreover , P14780 , P50281 , P01033 and P15692 were expressed at the highest levels in high-grade T-cell lymphomas . CONCLUSIONS : This study provides new information on the expression of different MMPs and P15692 in canine lymphoma , suggesting a possible correlation between different MMPs and P15692 , immunophenotype and prognosis . Are rofecoxib and celecoxib safer NSAIDS ? NSAIDs work by inhibiting the enzyme cyclo-oxygenase ( P36551 ) , responsible for prostaglandin synthesis . This enzyme exists in two isoforms , P23219 and P35354 . Inhibition of P23219 is thought to be the main cause of the gastrointestinal unwanted effects of NSAIDs , whilst inhibition of P35354 results in anti-inflammatory effects . [ symbol : see text ] DB00533 ( Vioxx -- MSD ) and [ symbol : see text ] celecoxib ( DB00482 -- Searle ) have been developed as selective inhibitors of P35354 . DB00533 is licensed for the symptomatic treatment of osteoarthritis , but not for rheumatoid arthritis . The manufacturer claims that " in clinical studies rofecoxib inhibits P35354 but not P23219 " , has " the power of high-dose NSAIDs -- diclofenac and ibuprofen " and " superior GI safety profile compared to conventional NSAIDs " . Celecoxib is licensed for symptom relief in osteoarthritis and rheumatoid arthritis . The manufacturer claims that celecoxib has " comparable efficacy and superior GI tolerability when compared to diclofenac or naproxen " . Here , we review rofecoxib and celecoxib and consider whether they are safer than conventional NSAIDs . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Celecoxib exhibits the greatest potency amongst cyclooxygenase ( P36551 ) inhibitors for growth inhibition of P35354 -negative hematopoietic and epithelial cell lines . P35354 ( P35354 ) is an important cellular target for both therapy and/or prevention of inflammatory disorders and cancer . The advent of selective P35354 inhibitors now allows a more precise and safer treatment approach . The screening of an array of cancer cell lines for growth inhibitory effects of P35354 -selective and -nonselective inhibitors , including celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) , produced two unanticipated findings . Firstly , the antiproliferative effects of celecoxib were noted to be of very similar magnitude for both hematopoietic and epithelial cancer cell lines . Most hematopoietic cell lines had no detectable P35354 expression by reverse transcription-PCR , and none expressed P35354 protein . In addition , P35354 -negative epithelial lines were found to have IC50s for celecoxib that were very similar to their P35354 + counterparts . Thus , important antiproliferative effects were observed that were independent of both the cell lineage and P35354 status . Secondly , it was also observed that P35354 inhibitor drugs , celecoxib and rofecoxib , with similar P35354 -selectivity and clinical efficacy for inflammatory indications , differed significantly in their in vitro antiproliferative effects on cancer cell lines . IC50s of 35-65 microM were observed for celecoxib across this entire panel of cell lines . Finally , no difference in the mode or degree of cytotoxicity was apparent between cell lines , because similar levels of apoptosis were observed in P35354 + and -negative cell lines after treatment with celecoxib , with correspondingly lower levels after rofecoxib treatment . These data are important in that they provide the first direct comparison of epithelial and hematopoietic cancer cell lines , as well as a direct comparison of the in vitro anticancer effects of the two clinically available P35354 inhibitors . Gemcitabine/ DB00762 /celecoxib in pancreatic cancer . Unresectable pancreatic cancer has few therapeutic options and a dismal prognosis . P35354 ( P35354 ) expression is increased at the RNA and protein levels in most human pancreatic cancers . The purpose of this trial was to determine whether the addition of a P35354 inhibitor to chemotherapy was beneficial . To date , 11 patients with inoperable pancreatic cancer have been treated with the combination of gemcitabine ( Gemzar ) , irinotecan ( Camptosar ) , and celecoxib ( DB00482 ) at 400 mg orally twice daily . Encouraging pain relief , improvement in performance status , and decreases in CA 19-9 and carcinoembryonic antigen levels have been observed . Quantification and characterisation of cyclo-oxygenase and lipid peroxidation inhibitory anthocyanins in fruits of Amelanchier . The levels of bioactive anthocyanins in the fruits of Amelanchier alnifolia , A. arborea and A. canadensis have been determined by HPLC . Cyanidin 3-galactoside ( 1 ) was present in the fresh fruit of the three species at concentrations of 155 , 390 and 165 mg/100 g , respectively . Cyanidin 3-glucoside ( 2 ) was present only in A. alnifolia and A. canadensis at concentrations of 54 and 48 mg/100 g , respectively . The anthocyanins were confirmed by LC- P19957 /MS and NMR studies . At 100 ppm , anthocyanin mixtures from the three species inhibited cyclo-oxygenase ( P36551 ) -1 and -2 enzymes at 66 and 67 % , 60 and 72 % , and 51 and 76 % , respectively . The positive controls used in the P36551 assays were aspirin , DB00482 and Vioxx at 180 , 1.67 and 1.67 ppm , respectively , and showed 74 and 69 % , 5 and 82 % and 0 and 85 % P23219 and P35354 inhibition , respectively . Anthocyanins 1 and 2 and cyanidin ( 3 ) inhibited P23219 enzyme 50.5 , 45.62 and 96.36 % , respectively , at 100 ppm , whereas P35354 inhibition was the highest for 3 at 75 % . In the lipid peroxidation inhibitory assay , anthocyanin mixtures at 10 ppm from the three species showed activities of 72 , 73 and 68 % , respectively , compared with 89 , 87 and 98 % for commercial anti-oxidants butylated hydoxyanisole , butylated hydroxytoluene , and tert-butylhydroxyquinone at 1.67 , 2.2 and 1.67 ppm , respectively . At 10 ppm , compounds 1-3 inhibited lipid peroxidation by 70 , 75 and 78 % , respectively . Epstein-Barr virus Zta-induced immunomodulators from nasopharyngeal carcinoma cells upregulate interleukin-10 production from monocytes . During lytic infection with Epstein-Barr virus ( EBV ) , several viral lytic proteins function to evade immune recognition or to actively suppress immune cells . An EBV lytic transactivator , Zta , induces an immunosuppressive cytokine interleukin 10 ( P22301 ) in B cells , but whether it regulates P22301 in the context of epithelial cells is unclear . In this study , we tested nasopharyngeal carcinoma ( NPC ) cell lines and found that Zta did not induce P22301 in these epithelial cells . Interestingly , the conditioned medium of Zta-expressing NPC cells enhanced P22301 production from monocytes . We further revealed that the P22301 -inducing effect involved at least two immunomodulators that were upregulated by Zta and secreted from NPC cells : granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) and prostaglandin E(2) ( PGE(2) ) . Zta was recruited to and activated the GM- P04141 promoter , thus upregulating GM- P04141 expression . Zta also activated the promoter of cyclooxygenase-2 ( P35354 ) , and Zta-induced P35354 increased downstream PGE(2) production . Cotreatment with GM- P04141 and PGE(2) synergistically induced P22301 production from monocytes . The P22301 -inducing effect of the Zta-conditioned medium was reduced when GM- P04141 or the P35354 /PGE(2) pathway was blocked . The conditioned medium of NPC cells with EBV lytic infection showed a similar increase of GM- P04141 and PGE(2) levels as well as the P22301 -inducing effect on monocytes , and knockdown of Zta abolished all the effects . Therefore , through Zta-induced immunomodulators , EBV lytic infection in NPC cells can direct bystander monocytes to produce P22301 , which may be a novel way of EBV to promote local immunosuppression . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . Functional interaction between p90Rsk2 and Emi1 contributes to the metaphase arrest of mouse oocytes . Vertebrate eggs arrest at metaphase of the second meiotic division before fertilization under the effect of a cytostatic factor ( P04141 ) . This arrest is established during oocyte maturation by the MAPK kinase module , comprised of Mos , MEK , MAPKs and p90Rsk . Maintenance of P04141 arrest at metaphase requires inhibitors of the anaphase-promoting complex ( P25054 ) like Emi1 , which sequesters the P25054 activator Cdc20 . Although it was proposed that the Mos pathway and Emi1 act independently , neither one alone is sufficient to entirely reproduce P04141 arrest . Herein we demonstrate that p90Rsk2 associates with and phosphorylates Emi1 upstream of the binding region for Cdc20 , thus stabilizing their interaction . Experiments in transfected cells and two-cell embryos indicate that Emi1 and p90Rsk2 cooperate to induce the metaphase arrest . Moreover , oocyte maturation was impaired by interfering with the interaction between p90Rsk2 and Emi1 or by RNA interference of Emi1 . Our results indicate that p90Rsk2 and Emi1 functionally interact during oocyte maturation and that the Mos pathway establishes P04141 activity through stabilization of an P25054 -inhibitory complex composed by Emi1 and Cdc20 before fertilization . Docking studies on NSAID/ P35354 isozyme complexes using contact statistics analysis . The selective inhibition of P35354 isozymes should lead to a new generation of NSAIDs with significantly reduced side effects ; e.g. celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . To obtain inhibitors with higher selectivity it has become essential to gain additional insight into the details of the interactions between P36551 isozymes-and NSAIDs . Although X-ray structures of P35354 complexed with a small number of ligands are available , experimental data are missing for two well-known selective P35354 inhibitors ( rofecoxib and nimesulide ) and docking results reported are controversial . We use a combination of a traditional docking procedure with a new computational tool ( Contact Statistics analysis ) that identifies the best orientation among a number of solutions to shed some light on this topic . A triple combination of atorvastatin , celecoxib and tipifarnib strongly inhibits pancreatic cancer cells and xenograft pancreatic tumors . Because K-Ras mutation and cyclooxygenase-2 ( P35354 ) overexpression are hallmarks of majority of pancreatic cancer patients , an approach to inhibit the progression and growth of pancreatic cancer using the simultaneous administration of agents that inhibit the function of both targets , should be considered . In the present study , we assessed the effects of atorvastatin ( Lipitor ) , celecoxib ( DB00482 ) and tipifarnib ( DB04960 ) on the growth of human pancreatic cancer . In the in vitro studies , we found that treatment of human pancreatic tumor cells with a combination of atorvastatin , celecoxib and tipifarnib had a stronger inhibitory effect on growth and a stronger stimulatory effect on apoptosis than each drug alone or for any combination of two drugs . We also found that treatment of Panc-1 cells with a combination of all three drugs strongly decreased the levels of phosphorylated Erk1/2 and Akt . In an animal model of xenograft tumors in severe combined immunodeficient ( SCID ) mice , we found that daily i.p. injections of a combination of atorvastatin , celecoxib and tipifarnib had a stronger inhibitory effect on the growth of the tumors in mice than each drug alone or for any combination of two drugs . The results of our study indicate that a combination of atorvastatin , celecoxib and tipifarnib may be an effective strategy for the treatment of pancreatic cancer . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . P19957 kinase activation and response to DB00072 Therapy : what 's neu with herceptin resistance ? DB00072 is an established therapy for women with breast cancers that overexpress P04626 . Despite its proven benefit in treating breast cancer , not all women derive benefit from this monoclonal antibody , and resistant disease can develop . In this issue of Cancer Cell , Berns et al. present evidence that activation of the P19957 kinase pathway , either via loss of the tumor suppressor P60484 or through oncogenic stimulation of P42336 , can mediate trastuzumab resistance . This study extends important work of others and forms the rationale for future investigations combining inhibitors of the P19957 kinase pathway in conjunction with trastuzumab therapy . Docosahexaenoic acid reduces amyloid-β(1-42) secretion in human AβPP-transfected CHO-cells by mechanisms other than inflammation related to PGE₂ . The effect of supplementation with the omega 3 polyunsaturated fatty acid ( n3 PUFA ) docosahexaenoic acid ( DB01708 ) on membrane composition and amyloid-β₁₋₄₂ ( Aβ₄₂ ) secretion was studied in human amyloid-β protein precursor-transfected Chinese Hamster Ovary ( CHO ) cells . Twenty-four hour incubation with a range of DB01708 concentrations resulted in a dose-dependent increase in membrane DB01708 and eicosapentaenoic acid content and a decrease in arachidonic acid content . In addition , DB01708 supplementation caused a dose-dependent reduction in the secreted Aβ₄₂ levels and resulted in a 4-8 fold decrease in extracellular prostaglandin E₂ ( PGE₂ ) levels . Tocopherol , which was added to DB01708 to prevent oxidation , may have contributed to the effect of DB01708 , since it slightly decreased extracellular Aβ₄₂ and PGE₂ levels when given alone . The addition of selective P35354 inhibitors DB00482 and curcumin to the culture medium resulted in a significant and comparable inhibition of PGE₂ release , but did not inhibit Aβ₄₂ secretion , and even significantly increased Aβ₄₂ production in this cell system . Together , the present data show that , whereas both DB01708 and P35354 inhibitors may reduce PGE₂ production , only DB01708 in the presence of tocopherol significantly reduced Aβ₄₂ production and concurrently changed membrane lipid composition in CHO cells . It is concluded that in this in vitro setting DB01708 reduced Aβ₄₂ secretion through membrane-related , but not PGE₂-related mechanisms . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Acute generalized exanthematic pustulosis : a case and an overview of side effects affecting the skin caused by celecoxib and other P35354 inhibitors reported so far . A 55-year-old woman who was treated for periarthritis humeroscapularis with celecoxib ( DB00482 ) developed a generalized pustular exanthema on the head and upper trunk , accompanied by fever , leukocytosis and increased erythrocyte sedimentation rate . The histological findings were subcorneal pustules , necrotic keratinocytes , edema in the upper dermis and polymorphic perivascular infiltrates . Four days after stopping celecoxib , the pustules disappeared without any treatment . Four weeks after disappearance of the skin lesions , celecoxib demonstrated a positive lymphocyte stimulation test . In this article , we present to our knowledge the first case of acute generalized exanthematic pustulosis caused by celecoxib , and we give an overview of the side effects affecting the skin caused by celecoxib and other cyclooxygenase type 2 inhibitors reported so far . Somatostatin preserved blood brain barrier against cytokine induced alterations : possible role in multiple sclerosis . Multiple sclerosis ( MS ) is an inflammatory neurological disorder associated with demyelination , impaired blood brain barrier ( BBB ) , axonal damage and neuronal loss . In the present study , we measured somatostatin ( P61278 ) and tumor necrosis factor-α ( P01375 -α ) like immunoreactivity in P04141 samples from MS and non-MS patients . We also examined the role of P61278 in cytokines and lipopolysaccharide ( LPS ) -induced damage to the BBB using human brain endothelial cells in culture . Most of the cerebrospinal fluid ( P04141 ) samples studied from definite MS patients exhibited lower somatostatin ( P61278 ) -like immunoreactivity and higher expression of P01375 -α in comparison to non-MS patients . Treatment of cells with cytokines and LPS blocked P61278 secretion and decreased P61278 expression . Human brain endothelial cells expressed all five somatostatin receptors ( SSTRs ) with increased expression of P30874 and 4 upon treatment with cytokines and LPS . Cytokines and LPS-induced disruption of the tight junction proteins Zonula occludens ( ZO-1 ) organization was restored in presence of P61278 , P30874 or P31391 selective agonists . Furthermore , inflammation induced changes in extracellular signal-regulated kinases ( P27361 /2 and Q13164 ) signaling and altered expression of endothelial and inducible nitric oxide synthase are modulated in presence of P61278 . These data indicate that decreased levels of P61278 contribute to failure of the BBB in MS . Celecoxib-induced upper gastrointestinal hemorrhage and ulceration . P35354 inhibitors are a new class of nonsteroidal anti-inflammatory drugs with a reported benefit of less gastric and duodenal ulceration and hemorrhage . We describe a 67-year-old man taking a higher than usual dose of celecoxib ( DB00482 ) for osteoarthritis with resultant gastric erosions , ulceration , and a significant gastrointestinal ( GI ) hemorrhage . A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . Effects of selective P35354 and 5- P28300 inhibition on prostaglandin and leukotriene synthesis in ductal pancreatic cancer in Syrian hamster . Selective inhibition of eicosanoid synthesis seems to decrease carcinogenesis , however , the effect on liver metastasis in pancreatic cancer is still unknown . Ductal pancreatic adenocarcinoma was chemically induced by weekly injection of N-nitrosobis-2-oxopropylamine ( BOP ) in Syrian hamster . Animals received selective inhibition of cyclooxygenase-2 ( DB00482 ) and P09917 ( DB00744 ) . In week 33 , hamsters were sacrificed and incidence of pancreatic carcinomas as well as liver metastases were examined . Furthermore , size and number of liver metastases per animal were determined and concentration of PGF1alpha , DB00917 and leukotrienes was measured in hepatic and pancreatic tissue . Combined therapy ( DB00482 + DB00744 ) significantly decreased incidence , number and size of liver metastases . Furthermore extra- and intrametastatic concentration of DB00917 was reduced by this treatment in hepatic tissue . Single Cox-2-inhibition ( DB00482 ) decreased intrametastatic hepatic PGF1alpha and DB00917 concentration while PGF1alpha concentration was reduced in non-metastatic liver ( nml ) . Moreover 5- P28300 -inhibition ( DB00744 ) decreased intrametastatic DB00917 concentration as well as PGF1alpha and DB00917 in nml . In pancreatic carcinomas highest LT-concentration was found after combined treatment and this therapy group was the only one revealing a significantly higher amount of LTs in carcinomas compared to tumour-free tissue . Hepatic LT-concentration was significantly lower in the control groups than in nml of the tumour groups . Combination of Cox-2-inhibition and 5-Lox-inhibition might be a suitable adjuvant therapy to prevent liver metastasis in human ductal pancreatic adenocarcinoma . Aggravated endoplasmic reticulum stress as a basis for enhanced glioblastoma cell killing by bortezomib in combination with celecoxib or its non-coxib analogue , 2,5-dimethyl-celecoxib . The proteasome inhibitor bortezomib ( Velcade ) is known to trigger endoplasmic reticulum ( ER ) stress via the accumulation of obsolete and damaged proteins . The selective cyclooxygenase-2 ( P35354 ) inhibitor celecoxib ( DB00482 ) causes ER stress through a different mechanism ( i.e. , by causing leakage of calcium from the ER into the cytosol ) . Each of these two mechanisms has been implicated in the anticancer effects of the respective drug . We therefore investigated whether the combination of these two drugs would lead to further increased ER stress and would enhance their antitumor efficacy . With the use of human glioblastoma cell lines , we show that this is indeed the case . When combined , bortezomib and celecoxib triggered elevated expression of the ER stress markers P11021 / P11021 and P35638 / P35638 , caused activation of c-Jun NH(2)-terminal kinase and ER stress-associated caspase-4 , and greatly increased apoptotic cell death . Small interfering RNA-mediated knockdown of the protective ER chaperone P11021 / P11021 further sensitized the tumor cells to killing by the drug combination . The contribution of celecoxib was independent of the inhibition of P35354 because a non-coxib analogue of this drug , 2,5-dimethyl-celecoxib ( Q6UXB2 ) , faithfully and more potently mimicked these combination effects in vitro and in vivo . Taken together , our results show that combining bortezomib with celecoxib or Q6UXB2 very potently triggers the ER stress response and results in greatly increased glioblastoma cytotoxicity . We propose that this novel drug combination should receive further evaluation as a potentially effective anticancer therapy . Immunohistochemical study of P15692 , angiopoietin 2 and their receptors in the neovascularization following microinjection of P13671 glioma cells into rat brain . BACKGROUND : Recent papers suggest that two angiogenic factors ( angiopoietin 2 and vascular endothelial growth factor ) cooperate in tumoral angiogenesis to support the growing tumor . The purpose of the present work was to demonstrate the existence of such cooperation in a longitudinal study of a brain tumor model during tumor growth by means of immunohistochemistry . MATERIALS AND METHODS : The study was performed on 31 rats bearing P13671 glioma . At different stages of tumor growth , the histological aspects were described and sections were immunostained for P15692 , Ang-2 and their receptors P17948 , P35968 and Tie-2 . Immunostaining was semi-quantitatively analyzed and the localization of immunostained cells was reported . RESULTS : Ang-2 and Tie-2 were detected in the endothelial cells of vessels surrounded by tumor cells , occuring early in our study , with immunostaining taking place from day 4 to day 24 . Immunostaining with P15692 ( on tumoral cells ) and P35968 ( on endothelial cells ) was present after 8 days of tumor growth . A clear increase of vessel density can be observed at the periphery of the tumors after 16 days of tumor growth . At that time , areas of necrosis were present in the tumor with concomitant P15692 and Ang-2 expression . CONCLUSION : The present study demonstrated cooperation between the early effect of Ang-2 and the secondary effect of P15692 to elaborate new vessels in a longitudinal study of experimental brain tumors . This study is favorable to the new model of tumor angiogenesis , with successive vessel cooption , regression and growth mediated by angiopoietins and P15692 . DB00759 derivative minocycline inhibits autophagy and inflammation in concanavalin-a-activated human hepatoma cells . Inhibition of soluble matrix metalloproteinase ( MMP ) activity is among the non-antibiotic cellular effects exerted by the anti-inflammatory tetracycline derivative minocycline . The impact of minocycline on the signal transduction functions of membrane-bound MMPs is however unknown . We assessed minocycline in a concanavalin-A ( ConA ) -activated human HepG2 hepatoma cell model , a condition known to increase the expression of membrane type-1 MMP ( MT-MMP ) and to trigger inflammatory and autophagy processes . We found that minocycline inhibited ConA-induced formation of autophagic acidic vacuoles , green fluorescent microtubule-associated protein 1 light chain 3 ( GFP-LC3 ) puncta formation , gene and protein expression of autophagy biomarker P10415 /adenovirus E1B 19 kDa interacting protein 3 ( Q12983 ) , invasion biomarker P50281 , and inflammation biomarker cyclooxygenase ( P36551 ) -2 . Gene silencing of P50281 abrogated ConA-induced formation of autophagic acidic vacuoles and ConA-induced expressions of Q12983 and P35354 . DB01017 was also shown to inhibit ConA-induced signal transducer and activator of transcription 3 ( P40763 ) phosphorylation as well as gene expression of Q8WY41 , a biomarker believed to colocalize with P50281 and the specific silencing of which further inhibited ConA-induced P40763 phosphorylation . Collectively , our data demonstrate that part of minocycline 's effects on autophagy could be exerted through the inhibition of P50281 signaling functions , which contribute to the autophagy and inflammatory phenotype of ConA-activated HepG2 cells . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Effects of P04626 amplicon size and genomic alterations of chromosomes 1 , 3 , and 10 on patient response to trastuzumab in metastatic breast cancer . DB00072 is widely used for advanced breast cancer patients with P04626 -amplified tumors . Nevertheless , over half of these patients do not have an objective response . One reason may be altered expression of genes that might compensate for P04626 inhibition . We previously mapped the gene-rich region of chromosome 17 telomeric to P04626 , and reported considerable variability in the telomeric extent of the P04626 amplicon . Here we examined whether the variable amplicon size may be associated with patient response to trastuzumab . In addition , we looked at associations between response and several signaling pathway-related genes unrelated to the P04626 amplicon , including Q9Y243 , P60484 , P42336 , and P35354 . In 35 patients with P04626 -amplified metastatic breast cancer , with 40 % overall response to trastuzumab , fluorescence in situ hybridization identified the telomeric extent of the P04626 amplicon and the status of the several pathway-related genes . Objective response strongly correlated with the telomeric amplicon size , with 62 % of patients with shorter amplicons responding , compared with only 7 % of patients with longer amplicons ( P = 0.0015 ) . Abnormal copy number of P35354 was marginally associated with objective response ( P = 0.066 ) , while abnormal copy numbers of two reference loci , 1q25 and the chromosome 10 centromere , were significantly associated with response . Pairwise combinations of copy number status of these loci and P04626 amplicon size provided stronger associations and identified a group of patients without responders . These results suggest that patient selection for trastuzumab may be improved by considering P04626 amplicon size and genomic status of the 1q25 , P35354 , and centromere 10 loci . [ A pop-up menu linked to a computerized drug prescribing system. Prescribing pattern 's feedback via a simple and quick method ] . It takes time for a GP to acquire sufficient experience of a new drug to be able to prescribe competently . This article describes a project studying the use of computerized records to afford a group of GP 's swift feedback on recently introduced drugs of special interest . In the south-east of Sweden a network of primary health care centers has been created in two neighboring counties . The pharmacies of the region are also taking part . When new drugs of particular interest are introduced , each participating GP will automatically see a pop-up menu , asking questions pertaining to each computer-assisted prescription . In the pharmacies , patients are given a questionnaire regarding their expectations with respect to the drug . In this way it will be possible to provide the individual GP swift feedback from a large number of colleagues and patients concerning the drug 's effectiveness in clinical practice . We have now been studying the P35354 inhibitors rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) . Results show that a pop-up menu used in this way provides the general practitioner quick feed-back on prescribing behavior as well as drug effectiveness in clinical practice . DB01259 -mediated cyclooxygenase-2 expression via epidermal growth factor receptor/ Q15717 interaction enhances the aggressiveness of triple-negative breast cancer cells . DB01259 , a dual epidermal growth factor receptor ( P00533 ) /human epidermal growth factor receptor 2 ( P04626 ) kinase inhibitor , showed clinical benefits in advanced P04626 -positive breast cancer patients . Because some triple-negative breast cancers ( TNBCs ) frequently overexpress P00533 , the antitumor activity of lapatinib in such diseases was also tested . However , the results showed a worse event-free survival rate . It remains unknown whether and how lapatinib elicits the aggressiveness of such cancer cells . In this study , our results demonstrated that lapatinib facilitated axillary and lung metastases of triple-negative MDA-MB-231 breast cancer cells without affecting their viability , leading to worse survival in orthotopic xenograft mice . The lapatinib-increased motility was attributed by the elevation of P00533 through the downregulation of microRNA-7 and by the subsequent overexpression of cyclooxygenase-2 ( P35354 ) . Strikingly , independent of its kinase activity , the elevated P00533 at least partly stabilized P35354 expression by enhancing the binding of Q15717 to P35354 mRNA . Our results suggest that lapatinib may increase the migration and invasion of MDA-MB-231 cells by upregulating P00533 and P35354 through the downregulation of microRNA-7 , providing a potential explanation for the worse clinical outcome of TNBC patients who receive lapatinib-based treatment . These findings also shed new light on the molecular mechanism of P35354 mRNA stabilization by P00533 in a kinase-independent manner . A real time quantitative PCR analysis and correlation of P23219 and P35354 enzymes in inflamed dental pulps following administration of three different NSAIDs . Dental pain is encountered daily by clinicians . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) commonly used for pain management are traditionally cyclooxygenase-1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) inhibitors , and more recently selective P35354 inhibitors . This study was designed to identify and quantify P23219 and P35354 gene expression level in inflamed rat molar pulps after administration of three NSAIDs : DB00482 , Vioxx , and DB01050 . Fifty male Wistar rats had their first and second molar pulps exposed and sealed with Cavit for 4 days . Rats were randomly divided into the three drug groups and two control groups . RNA was isolated from the rat pulps . Real Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction assay , a relatively new PCR technique , was used to quantify P23219 and P35354 mRNA . Statistical analysis demonstrated no significant differences in P23219 and P35354 levels among the drug groups . However , Vioxx and DB01050 significantly reduced P35354 expression levels compared to inflamed ( positive control ) pulps ( p < 0.05 ) . Effect of celecoxib on capecitabine-induced hand-foot syndrome and antitumor activity . We hypothesized that hand-foot syndrome is an inflammatory phenomenon mediated by the overexpression of cyclooxygenase 2 ( P35354 ) . Therefore , a specific P35354 inhibitor such as celecoxib ( DB00482 ) could attenuate both the incidence and severity of hand-foot syndrome . We undertook a retrospective study comparing the incidences of hand-foot syndrome in 67 patients with metastatic colorectal cancer who took capecitabine ( DB01101 ) with or without celecoxib . Surprisingly , celecoxib seemed to attenuate capecitabine-induced diarrhea as well . DB01101 /celecoxib was also associated with increased tumor response , proportion of stable disease ( 62.5 % vs 22.8 % , P = .001 ) , and increase in median time to tumor progression ( 6 vs 3 months , P = .002 ) compared with capecitabine alone , despite the fact that patients on capecitabine/celecoxib had less favorable disease characteristics ( age , performance status , and prior chemotherapies ) . Overexpression of P35354 , implicated in promoting angiogenesis , enhanced tumor invasiveness , evasion of apoptosis , and immune suppression , is a bona fide molecular target for many solid tumors , including colorectal cancer . Combining capecitabine with celecoxib in the treatment of colorectal cancer has strong preclinical rationales . A prospective study is being designed to evaluate capecitabine and celecoxib with or without epidermal growth factor receptor antagonist ZD1839 in the frontline treatment of metastatic colorectal cancer . These regimens under study are orally based and may significantly impact quality of life in the frontline treatment of metastatic colorectal cancer . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Current application of selective P35354 inhibitors in cancer prevention and treatment . The multistep process of carcinogenesis , which can take many years , provides many opportunities for intervention to inhibit disease progression . Effective chemoprevention agents may reduce the risk of cancer by inhibiting the initiation stage of carcinoma through induction of apoptosis or DNA repair in cells harboring mutations , or they may act to prevent promotion of tumor growth . Similarly , chemoprevention may entail blocking cancer progression to an invasive phenotype . Over the past decade , in vitro , preclinical , and clinical data have supported the hypothesis that cyclooxygenase ( P36551 ) -2 plays a central role in oncogenesis and that treatment with P35354 inhibitors offers an effective chemoprevention strategy , as exemplified by the activity of celecoxib ( DB00482 ) in familial adenomatous polyposis . These P35354 data have contributed to initiation of clinical trials testing P35354 inhibitors for the chemoprevention of a wide variety of cancers that overexpress P35354 . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . The role of celecoxib in Rad51 expression and cell survival affected by gefitinib in human non-small cell lung cancer cells . Celecoxib ( DB00482 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor and gefitinib ( DB00317 (R) , ZD1839 ) is a selective epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor for human non-small cell lung cancer ( NSCLC ) . The addition of celecoxib to gefitinib to prolong the survival of patients with NSCLC still remains controversial and needs to be investigated . The Rad51 protein is essential for homologous recombination repair , and is overexpressed in chemo- or radioresistant carcinomas . In this study , we characterize the role of celecoxib in the cytotoxicity , P27361 /2 activation and Rad51 expression affected by gefitinib in NSCLC cells . We show that celecoxib can enhance the cytotoxicity induced by gefitinib in NSCLC cells . Treatment with celecoxib alone has no effect on the P27361 /2 activation , Rad51 mRNA and protein levels , however , combined treatment with gefitinib results in a significant reduction of phospho- P27361 /2 and Rad51 protein levels , and triggers the degradation of Rad51 via a 26S proteasome-dependent pathway . Expression of constitutively active Q02750 /2 vectors ( Q02750 /2-CA ) significantly rescues the decreased P27361 /2 activity , and restores Rad51 protein levels and cell survival under co-treatment with gefitinib and celecoxib . Furthermore , blocking P27361 /2 activation by U0126 ( Q02750 /2 inhibitor ) and knocking down Rad51 expression by transfection with small interfering RNA of Rad51 can enhance the cytotoxicity of celecoxib .

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DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Herpes simplex virus infection causes cellular beta-amyloid accumulation and secretase upregulation . It is uncertain whether environmental factors contribute to the formation of senile plaques and neurofibrillary tangles , the abnormal features that define the Alzheimer 's disease ( AD ) brain . We previously proposed that herpes simplex virus type 1 ( HSV1 ) is a strong risk factor for AD when it is present in the brains of people who possess the type 4 allele of the apolipoprotein E gene ( P02649 -epsilon4 ) ; however a direct biochemical link between viral infection and the development of the AD pathological features has never previously been examined . Here we show that infection of cultured neuronal and glial cells with HSV1 leads to a dramatic increase in the intracellular levels of beta-amyloid ( Abeta ) 1-40 and 1-42 , whilst levels of amyloid precursor protein ( P05067 ) in cells decrease . Similarly , Abeta1-42 deposits are present in mouse brain after HSV1 infection . In the cultured cells the mechanism involves increased Abeta production , rather than merely greater retention of cellular Abeta , as levels of beta-site P05067 -cleaving enzyme ( P56817 -1 ) and of nicastrin , a component of gamma-secretase , both increase in HSV1-infected cells . These novel data show that HSV1 can directly contribute to the development of senile plaques . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . Novel systemic markers for patients with Alzheimer disease ? - a pilot study . Almost 2 % of the population of western industrialized countries are affected by Alzheimer 's disease ( AD ) . Nevertheless the pathogenetic process leading to this neurodegenerative disease is widely unknown . Thus , we focus on novel pathophysiological aspects of AD . We hypothesize that AD patients reveal increased levels of peripheral blood mononuclear cells ( PBMCs ) expressing proinflammatory ( P35354 , P01375 , P25942 ) , proapoptotic ( P09874 ) , adhesion-relevant ( P28907 ) or AD associated ( C99 , P56817 , P49768 ) proteins as well as elevated proinflammatory biochemical plasma parameters . Therefore , PBMCs of AD patients and age-matched control subjects were studied by two color fluorescence-activated cell sorter ( FACS ) analysis . Furthermore , concentration of plasma oxidized low-density lipoprotein ( oxLDL ) and P01375 were measured by enzyme-linked immunosorbent assay ( ELISA ) . We found a significantly increased percentage of P01375 , P35354 , P09874 , P28907 , C99 or presenilin-1 positive PBMCs in AD patients compared with healthy subjects . FACS analyses revealed that the percentage of C99 or presenilin-1 positive PBMCs , which also express P01375 , P35354 , P09874 or P28907 is also increased in AD patients . Additionally , AD patients had significantly increased plasma oxLDL and P01375 levels . Furthermore , we found positive correlations between plasma oxLDL or P01375 concentrations and the percentage of TNFalpha+ , P35354 + or P09874 + , as well as P49768 + , C99+ or P56817 + PBMCs . Our findings suggest that immunocytological investigations , based on immunophenotyping of AD relevant proteins combined with measurement of proinflammatory , proapoptotic and adhesion-relevant proteins in PBMCs may provide more insight into the pathophysiology of AD . α-Asarone Ameliorates Memory Deficit in Lipopolysaccharide-Treated Mice via Suppression of Pro-Inflammatory Cytokines and Microglial Activation . α-Asarone exhibits a number of pharmacological actions including neuroprotective , anti-oxidative , anticonvulsive , and cognitive enhancing action . The present study investigated the effects of α-asarone on pro-inflammatory cytokines mRNA , microglial activation , and neuronal damage in the hippocampus and on learning and memory deficits in systemic lipopolysaccharide ( LPS ) -treated C57BL/6 mice . Varying doses of α-asarone was orally administered ( 7.5 , 15 , or 30 mg/kg ) once a day for 3 days before the LPS ( 3 mg/kg ) injection . α-Asarone significantly reduced P01375 -α and IL-1β mRNA at 4 and 24 hours after the LPS injection at dose of 30 mg/kg . At 24 hours after the LPS injection , the loss of P00915 neurons , the increase of TUNEL-labeled cells , and the up-regulation of P56817 expression in the hippocampus were attenuated by 30 mg/kg of α-asarone treatment . α-Asarone significantly reduced Iba1 protein expression in the hippocampal tissue at a dose of 30 mg/kg . α-Asarone did not reduce the number of Iba1-expressing microglia on immunohistochemistry but the average cell size and percentage areas of Iba1-expressing microglia in the hippocampus were significantly decreased by 30 mg/kg of α-asarone treatment . In the Morris water maze test , α-asarone significantly prolonged the swimming time spent in the target and peri-target zones . α-Asarone also significantly increased the number of target heading and memory score in the Morris water maze . The results suggest that inhibition of pro-inflammatory cytokines and microglial activation in the hippocampus by α-asarone may be one of the mechanisms for the α-asarone-mediated ameliorating effect on memory deficits . Calpain activation promotes P56817 expression , amyloid precursor protein processing , and amyloid plaque formation in a transgenic mouse model of Alzheimer disease . Abnormal activation of calpain is implicated in synaptic dysfunction and participates in neuronal death in Alzheimer disease ( AD ) and other neurological disorders . Pharmacological inhibition of calpain has been shown to improve memory and synaptic transmission in the mouse model of AD . However , the role and mechanism of calpain in AD progression remain elusive . Here we demonstrate a role of calpain in the neuropathology in amyloid precursor protein ( P05067 ) and presenilin 1 ( P49768 ) double-transgenic mice , an established mouse model of AD . We found that overexpression of endogenous calpain inhibitor calpastatin ( CAST ) under the control of the calcium/calmodulin-dependent protein kinase II promoter in P05067 / P49768 mice caused a remarkable decrease of amyloid plaque burdens and prevented Tau phosphorylation and the loss of synapses . Furthermore , CAST overexpression prevented the decrease in the phosphorylation of the memory-related molecules CREB and P29323 in the brain of P05067 / P49768 mice and improved spatial learning and memory . Interestingly , treatment of cultured primary neurons with amyloid-beta ( Abeta ) peptides caused an increase in the level of beta-site P05067 -cleaving enzyme 1 ( P56817 ) , the key enzyme responsible for P05067 processing and Abeta production . This effect was inhibited by CAST overexpression . Consistently , overexpression of calpain in heterologous P05067 expressing cells up-regulated the level of P56817 and increased Abeta production . Finally , CAST transgene prevented the increase of P56817 in P05067 / P49768 mice . Thus , calpain activation plays an important role in P05067 processing and plaque formation , probably by regulating the expression of P56817 . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . DB08877 for the treatment of primary myelofibrosis . PURPOSE : The pharmacology , pharmacokinetics , pharmacogenomics , clinical efficacy , and safety profile of ruxolitinib for the treatment of primary myelofibrosis are reviewed . SUMMARY : DB08877 , an oral tyrosine kinase inhibitor that targets the Janus-associated kinases ( JAKs ) 1 and 2 , has been recently approved for the treatment of patients with intermediate- or high-risk myelofibrosis . Unlike previous treatment options for patients with myelofibrosis , ruxolitinib offers a targeted therapy option for these patients who often suffer with severe and debilitating symptoms associated with the disease process . After oral administration , ruxolitinib is rapidly absorbed and can be given without regard to meals . DB08877 is primarily metabolized by the cytochrome P-450 ( CYP ) 3A4 isoenzyme system ; therefore , if concomitant use with a strong P08684 inhibitor is unavoidable , an initial dosage reduction is warranted . Two Phase III randomized trials comparing ruxolitinib to either placebo or best available therapy found a rapid and sustained response in the reduction of spleen size and improvements in constitutional symptoms and quality of life , with one study demonstrating an improvement in overall survival . The most commonly reported serious adverse effects of ruxolitinib are anemia and thrombocytopenia . DB08877 is administered as an oral tablet given twice daily , with the initial starting dosage based on the baseline platelet count . Dosage reductions are based on the development of thrombocytopenia . CONCLUSION : By directly targeting both P23458 and O60674 through small-molecule inhibition , ruxolitinib elicits a reduction in splenomegaly and disease-related symptoms in patients with intermediate- or high-risk myelofibrosis while maintaining an acceptable toxicity profile and a low treatment-discontinuation rate . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . P01579 -induced P56817 expression is mediated by activation of O60674 and P27361 /2 signaling pathways and direct binding of P42224 to P56817 promoter in astrocytes . P56817 ( P56817 ) is an essential enzyme for the production of beta amyloid . Since we found that injection of interferon-gamma ( P01579 ) into young mouse brains increased P56817 expression in astrocytes , we investigated molecular mechanisms underlying this process by cloning a putative P56817 promoter . P56817 promoter activity was differentially regulated by P01579 in a region specific manner and down-regulated by an inhibitor of O60674 ( O60674 ) . A dominant negative mutant of signal transducer and activator of transcription 1 ( P42224 ) expression suppressed P56817 promoter activity , and this was rescued by transfecting wild type P42224 . Electrophoretic mobility shift assay and promoter activity assays indicated that P42224 binds directly to the putative P42224 binding sequence of P56817 promoter . Because P01579 treatment induced P42224 phosphorylation , we examined whether the expression of a suppressor of cytokine signaling ( Q9NSE2 ) , negative regulator of O60674 , suppresses P56817 promoter activity . The results show that O15524 or O14543 expression suppressed P56817 promoter by blocking phosphorylation of Tyr701 residue in P42224 . Also , because P01579 treatment specifically potentiated extracellular signal regulated Q96HU1 kinase ( P29323 ) 1/2 activation , pretreatment of mitogen-activated or extracellular signal-regulated protein kinase ( MEK ) inhibitor , PD98059 , significantly attenuated P01579 -induced P56817 promoter activity and protein expression through blocking phosphorylation of Ser727 residue in P42224 , suggesting that P27361 /2 is associated with P01579 -induced P42224 signaling cascade . Taken together , our results suggest that P01579 activates O60674 and P27361 /2 and then phosphorylated P42224 binds to the putative P42224 binding sequences in P56817 promoter region to modulate P56817 protein expression in astrocytes . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Accumulation of autophagosomes contributes to enhanced amyloidogenic P05067 processing under insulin-resistant conditions . Alzheimer disease ( AD ) is sometimes referred to as type III diabetes because of the shared risk factors for the two disorders . P01308 resistance , one of the major components of type II diabetes mellitus ( T2DM ) , is a known risk factor for AD . P01308 resistance increases amyloid-β peptide ( Aβ ) generation , but the exact mechanism underlying the linkage of insulin resistance to increased Aβ generation in the brain is unknown . In this study , we investigated the effect of insulin resistance on amyloid β ( A4 ) precursor protein ( P05067 ) processing in mice fed a high-fat diet ( HFD ) , and diabetic db/db mice . We found that insulin resistance promotes Aβ generation in the brain via altered insulin signal transduction , increased P56817 /β-secretase and γ-secretase activities , and accumulation of autophagosomes . Using an in vitro model of insulin resistance , we found that defects in insulin signal transduction affect autophagic flux by inhibiting the mechanistic target of rapamycin ( P42345 ) pathway . The insulin resistance-induced autophagosome accumulation resulted in alteration of P05067 processing through enrichment of secretase proteins in autophagosomes . We speculate that the insulin resistance that underlies the pathogenesis of T2DM might alter P05067 processing through autophagy activation , which might be involved in the pathogenesis of AD . Therefore , we propose that insulin resistance-induced autophagosome accumulation becomes a potential linker between AD and T2DM . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . P01579 stimulates beta-secretase expression and sAPPbeta production in astrocytes . Neurons , but not astrocytes , are known as the major source of Abeta , because astrocytes express low levels of putative beta-secretase ( P56817 ) . Astrocytes near senile plaque cores show enhanced levels of P56817 protein expression , however , suggesting that astrocytes can contribute to Abeta production under pathological conditions . To investigate factors that stimulate P56817 protein expression in astrocytes , we tested the effects of interleukin-1beta ( IL-1beta ) and interferon-gamma ( P01579 ) on P56817 protein expression in U373MG astrocytoma cells and primary astrocyte cultures from Tg2576 mouse brains . P56817 protein expression and sAPPbeta production were dramatically increased , without changes in holo P05067 levels , following P01579 treatment in both cell types . AG490 , which is a blocker of P01579 -induced P35610 signaling , decreased P01579 -induced P56817 protein expression and sAPPbeta production in a dose-dependent manner . These results show that astrocytes are capable of expressing P56817 and producing sAPPbeta in response to certain stimulating factors , and P01579 is one such factor .

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interacts_with DB00181?

multiple_choice

P13987 DX , a double lipoxygenase product of DB01708 , inhibits both ROS production in human neutrophils and cyclooxygenase activities . Neutrophils play a major role in inflammation by releasing large amounts of reactive oxygen species ( ROS ) produced by NADPH oxidase ( NOX ) and myeloperoxidase ( P05164 ) . This ROS overproduction is mediated by phosphorylation of the NOX subunits in an uncontrolled manner . Therefore , targeting neutrophil subunits would represent a promising strategy to moderate NOX activity , lower ROS , and other inflammatory agents , such as cytokines and leukotrienes , produced by neutrophils . For this purpose , we investigated the effects of protectin DX ( DB06813 ) -a docosahexaenoic acid di-hydroxylated product which inhibits blood platelet aggregation-on neutrophil activation in vitro . We found that DB06813 decreases ROS production , inhibits NOX activation and P05164 release from neutrophils . We also confirm , that DB06813 is an anti-aggregatory and anti-inflammatory agent by inhibiting both cyclooxygenase-1 and -2 ( P23219 and P35354 , E.C. 1.14.99.1 ) as well as P35354 in lipopolysaccharides-treated human neutrophils . However , DB06813 has no effect on the P09917 pathway that produces the chemotactic agent leukotriene B4 ( LTB4 ) . Taken together , our results suggest that DB06813 could be a protective agent against neutrophil invasion in chronic inflammatory diseases . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Screening for candidate gene regions in narcolepsy using a microsatellite based approach and pooled DNA . Narcolepsy is a complex sleep disorder characterized by excessive daytime sleepiness and cataplexy . Mutations in genes of the hypocretin ( orexin ) neurotransmitter system cause narcoleptic symptoms in animal models . The absence of hypocretin in the cerebrospinal fluid of human patients is hypothesized to originate from destruction of hypocretinergic cells in the hypothalamus , the cause of which remains unknown . Due to strong HLA association autoimmune models of narcolepsy pathogenesis are still mostly favored . Genetic predisposition factors other than HLA are likely to play a role in causing the disorder . We screened three sets of gene regions ( n=254 ) for association with narcolepsy using a microsatellite based approach and pooled DNA : genes related to immunity , particularly apoptosis ; genes related to regulation of circadian rhythmicity ; genes coding for several factors of neurotransmission . In relation to apoptosis an association was found for the Q99933 gene region . Interestingly , microsatellites representing four genomic regions related to neurotransmission revealed association with narcolepsy : P21964 , P14416 , Q9UBS5 , and P28223 . These results , although exploratory and still to be confirmed in independent samples , support a complex pathogenetic model for narcolepsy , including disturbances of neurotransmission rather than involvement of autoimmunity . DB01095 inhibits growth and alters the malignant phenotype of the P13671 glioma cell line . BACKGROUND : DB01095 is a member of the family of P04035 inhibitors ( statins ) extensively used in medical practice . Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development . The aim of the present study was to investigate the anti-cancer potential of fluvastatin in P13671 rat malignant glioma cells . METHODS : First , the effects of fluvastatin on cell viability ( MTT assay ) , proliferation ( BrdU assay ) , cell morphology , and cytoskeleton were examined . Subsequently , its effect on extracellular signal regulated kinase 1 and 2 ( P27361 /2 ) and P45983 and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 and P15692 was determined using a wound-healing assay and ELISA test , respectively . RESULTS : The results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor P13671 cells ( IC(50) = 8.6 μM , 48 h ) , but did not inhibit the growth of normal neuronal cells . The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm , chromatin condensation , and nucleus breakdown . CONCLUSION : The inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p- P27361 /2 expression , upregulation of p- P45983 /2 , and reduction in the P14780 and P15692 concentrations in culture media . The high anticancer ( antiproliferative , proapoptotic , antiinvasive ) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma . [ Differential gene expression in nasopharyngeal carcinoma cell with reduced and normal expression of 6A8 alpha-mannosidase ] . OBJECTIVE : To detect the differential display of mRNA expression between human nasopharyngeal carcinoma cell CNE-2L2 with reduced malignancy caused by transduction of a DNA antisense to 6A8 alpha-mannosidase cDNA ( AS cell ) and the wild type cell ( W cell ) . METHODS : Differential display of mRNA expression was analyzed using DNA microarray analysis . The datasets were confirmed by Northern blotting and RT-PCR . RESULTS : Out of the 1069 genes analyzed , 34 genes were up-regulated in AS cells relative to W cells . Conversely , 42 genes were down-regulated . The genes , up-regulation of which might have suppressive effect on tumor malignant behaviors , were P130 mRNA for 130K protein , TGF-betaIIR alpha , Q9UBS5 , P36897 , Q13829 , STANIN , E-CADHERIN , P35221 and 2 , P48378 , TMPO , etc . The genes , down-regulation of which might have suppressive effect on tumor malignant behaviors , were P16070 , Q92597 , P01137 , P46782 , LEGUMAIIN , P35520 , P13987 , P09661 , etc . The microarray datasets were confirmed by Northern blot and RT-PCR analysis . CONCLUSIONS : In comparison to the W cell , AS cell has up-regulation of 34 genes and down-regulation of 42 genes . Changes of the gene expression may play a role in the malignancy reduction of AS cell . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Type B gamma-aminobutyric acid receptors modulate the function of the extracellular Ca2+-sensing receptor and cell differentiation in murine growth plate chondrocytes . Extracellular calcium-sensing receptors ( CaRs ) and metabotropic or type B gamma-aminobutyric acid receptors ( GABA-B-Rs ) , two closely related members of family C of the G protein-coupled receptor superfamily , dimerize in the formation of signaling and membrane-anchored receptor complexes . We tested whether CaRs and two GABA-B-R subunits ( Q96GN5 and R2 ) are expressed in mouse growth plate chondrocytes ( GPCs ) by PCR and immunocytochemistry and whether interactions between these receptors influence the expression and function of the CaR and extracellular Ca(2+)-mediated cell differentiation . Both CaRs and the Q9UBS5 and -R2 were expressed in the same zones of the growth plate and extensively colocalized in intracellular compartments and on the membranes of cultured GPCs . The Q9UBS5 co-immunoprecipitated with the CaR , confirming a physical interaction between the two receptors in GPCs . In vitro knockout of Q9UBS5 genes , using a Cre-lox recombination strategy , blunted the ability of high extracellular Ca(2+) concentration to activate phospholipase C and P27361 /2 , suppressed cell proliferation , and enhanced apoptosis in cultured GPCs . In GPCs , in which the Q9UBS5 was acutely knocked down , there was reduced expression of early chondrocyte markers , aggrecan and type II collagen , and increased expression of the late differentiation markers , type X collagen and osteopontin . These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs , potentially by altering the function of CaRs .

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interacts_with DB00208?

multiple_choice

A critical appraisal of the functional evolution of Q9H244 antagonists as antiplatelet drugs . Q9H244 receptor mediated inhibition of platelet aggregation is one of the most explored and exploited pathways in antiplatelet drug therapy to prevent ischemic events in patients undergoing percutaneous coronary intervention ( P05154 ) for the treatment of the acute coronary syndrome ( ACS ) . DB00208 , DB00758 , Prasugrel , DB08816 , DB06441 and Elinogrel are the Q9H244 inhibitors that act as antiplatelet drugs . In this review , the features of these drugs and the factors reported to be responsible for drug resistance or drug ineffectiveness were described . The features like drug metabolism , reversible or irreversible binding of drugs to their target protein and the mode of administration were observed to evolve along with the antiplatelet drugs . These features also include the drug-drug interactions , the pharmacogenetics and pharmacodynamics of Q9H244 inhibitors . We attempted to critically analyze how the desirable features were met by the Q9H244 inhibitors in the course of time . This review provides an overview of the evolution of Q9H244 inhibitors and may guide the researchers to develop better antiplatelet drugs in the future . Localisation of P47900 and P51582 receptors in dorsal root , nodose and trigeminal ganglia of the rat . The presence and distribution of P2Y ( nucleotide ) receptor subtypes in rat sensory neurons has been investigated . RT-PCR showed that P2Y(1) , P2Y(2) , P2Y(4) and P2Y(6) receptor mRNA is expressed in sensory ganglia [ dorsal root ganglion ( Q86YR7 ) , nodose ganglion ( NG ) and trigeminal ganglion ( TG ) ] . The regional and cellular distribution of P2Y(1) and P2Y(4) receptor proteins in these ganglia was investigated using immunohistochemistry . P2Y(1) polyclonal antibodies stained over 80 % of the sensory neurons , particularly the small-diameter ( neurofilament-negative ) neurons . The P2Y(4) receptor antibody stained more medium- and large- ( neurofilament-positive ) diameter neurons than small-diameter neurons . P2Y(1) and P2Y(4) receptor immunoreactivity ( P2Y(1)-IR and P2Y(4)-IR ) was often coexpressed with P2X(3) receptor immunoreactivity ( P2X(3)-IR ) in subpopulations of neurons . Double immunohistochemistry showed that 73-84 % of P2X(3) receptor-positive neurons also stained for the P2Y(1) receptor in Q86YR7 , TG and NG while only 25-35 % also stained for the P2Y(4) receptor . Subpopulations of P2Y(1)-IR neurons were coexpressed with NF200 , P80511 and IB(4) ; most P2Y(4)-IR neurons were coexpressed with NF200 , while only a few neurons were coexpressed with P80511 ( 10-20 % ) or with IB(4) ( 1-2 % ) . The results suggest that P2Y as well as P2X receptor subtypes contribute to purinergic signalling in sensory ganglia . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . DB00945 antagonizes the cytotoxic effect of methotrexate in lung cancer cells . DB00563 ( MTX ) has been widely used for the treatment of cancer and rheumatoid arthritis ( RA ) . DB00945 ( ASA ) is a non-selective cyclooxygenase ( P36551 ) inhibitor that contributes to the treatment of inflammatory conditions such as RA . It has been observed that the antitumor effect of ASA can be attributed to inhibition of cell cycle progression , induction of apoptosis and inhibition of angiogenesis . In the present study , we revealed that the treatment with a combination of MTX and ASA resulted in antagonism of the cytotoxic effect as demonstrated by P50991 and colony formation assays . ASA alleviated the MTX-mediated S phase accumulation and recovered the P55008 phase . MTX-mediated accumulation of the S phase marker cyclin A was also alleviated by ASA . Notably , FAS protein levels were upregulated by MTX in A549 cells . The antagonism of MTX efficacy caused by ASA was accompanied by altered expression of caspase-3 , Bcl-2 and FAS but not dihydrofolate reductase ( P00374 ) . This suggests that the alteration of caspase-3 , Bcl-2 and FAS was involved in the antagonism between ASA and MTX . Exogenously added folic acid reversed the MTX-mediated P00374 inhibition following either MTX or MTX + ASA treatments . Most importantly , we demonstrated for the first time that the commonly used non-steroidal anti-inflammatory drug for headache ASA and possibly other P23219 /2 inhibitors can produce a strong antagonistic effect on the growth inhibition of lung cancer cells when administered in combination with MTX . The clinical implication of our finding is obvious , i.e. , the clinical efficacy of MTX therapy can be compromised by ASA and their concomitant use should be avoided . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . Q9H244 receptor signalling towards P31749 proceeds through P08069 cross-talk and requires activation of Src , Pyk2 and Rap1 . Previously it was shown that stimulation of the Q9H244 receptor activates P31749 signalling in P13671 glioma cells [ K. Van Kolen and H. Slegers , J. Neurochem. 89 , 442. ] . In the present study , the mechanisms involved in this response were further elucidated . In cells transfected with the Gbetagamma-scavenger beta- O14965 / P25098 or Rap1GAPII , stimulation with 2MeSADP failed to enhance P31749 phosphorylation demonstrating that the signalling proceeds through Gbetagamma-subunits and Rap1 . Moreover , Rap1-GTP pull-down assays revealed that Q9H244 receptor stimulation induced a rapid activation of Rap1 . Treatment of cells with the Ca2+ chelator BAPTA-AM and inhibition of Src and O14939 with Q99463 or 1-butanol , respectively , abrogated Q9H244 receptor-mediated activation of Rap1 and P31749 . In addition inhibition of PKCzeta decreased basal and 2MeSADP-stimulated phosphorylation of P31749 indicating a role for this PKC isoform in P31749 signalling . Although the increased P31749 phosphorylation was abolished in the presence of the P08069 tyrosine kinase inhibitor AG 1024 , 2MeSADP did not significantly increase receptor phosphorylation . Nevertheless , phosphorylation of a 120 kDa P08069 -associated protein was observed . The latter protein was identified by MALDI-TOF/TOF-MS as the proline-rich tyrosine kinase 2 ( Pyk2 ) that co-operates with Src in a O14939 -dependent manner . Consistent with the signalling towards Rap1 and P31749 , activation of Pyk2 was abrogated by Ca2+ chelation , inhibition of O14939 and P08069 tyrosine kinase activity . In conclusion , the data reveal a novel type of cross-talk between Q9H244 and P05019 receptors that proceeds through Gbetagamma- , Ca2+-and O14939 -dependent activation of the Pyk2/Src pathway resulting in GTP-loading of Rap1 required for an increased P31749 phosphorylation . A novel sulindac derivative inhibits lung adenocarcinoma cell growth through suppression of Akt/ P42345 signaling and induction of autophagy . Nonsteroidal anti-inflammatory drugs such as sulindac sulfide have shown promising antineoplastic activity in multiple tumor types , but toxicities resulting from P36551 inhibition limit their use in cancer therapy . We recently described a N,N-dimethylethyl amine derivative of sulindac sulfide , sulindac sulfide amide ( SSA ) , that does not inhibit P23219 or -2 , yet displays potent tumor cell growth-inhibitory activity . Here , we studied the basis for the growth-inhibitory effects of SSA on human lung adenocarcinoma cell lines . SSA potently inhibited the growth of lung tumor cells with IC50 values of 2 to 5 μmol/L compared with 44 to 52 μmol/L for sulindac sulfide . SSA also suppressed DNA synthesis and caused a G0- P55008 cell-cycle arrest . SSA-induced cell death was associated with characteristics of autophagy , but significant caspase activation or PARP cleavage was not observed after treatment at its IC50 value . siRNA knockdown of Atg7 attenuated SSA-induced autophagy and cell death , whereas pan-caspase inhibitor ZVAD was not able to rescue viability . SSA treatment also inhibited Akt/ P42345 signaling and the expression of downstream proteins that are regulated by this pathway . Overexpression of a constitutively active form of Akt was able to reduce autophagy markers and confer resistance to SSA-induced cell death . Our findings provide evidence that SSA inhibits lung tumor cell growth by a mechanism involving autophagy induction through the suppression of Akt/ P42345 signaling . This unique mechanism of action , along with its increased potency and lack of P36551 inhibition , supports the development of SSA or related analogs for the prevention and/or treatment of lung cancer . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Involvement of platelet activation by Q9H244 receptor in the development of transplant arteriosclerosis in mice . BACKGROUND : Although activated platelets influence inflammation by intraplatelet mediators in transplantation , their mechanism of involvement in the progression of transplant arteriosclerosis has not yet been elucidated . We , therefore , investigated this question using Q9H244 receptor knockout ( KO ) mice , in which platelets can not be aggregated by adenosine diphosphate stimulation . METHODS : Carotid arteries from 129X1 mice were orthotopically transplanted into wild-type or KO mice in a minor antigen(s)-mismatched strain combination . No immunosuppression was used . Grafts were harvested at 7 , 14 , 28 , and 56 days after transplantation for morphometry and immunohistology . Fluorescence-activated cell sorting and quantitative real-time reverse-transcriptase polymerase chain reaction were performed at 7 and 14 days after transplantation . RESULTS : The intima/media ratio of grafts in KO mice was significantly reduced compared with wild-type mice at 14 , 28 , and 56 days after transplantation . Fluorescence-activated cell sorting analysis showed a significant reduction of platelet CD154 expression and platelet-leukocyte aggregates in KO mice at 14 days after transplantation . Additionally , levels of intercellular adhesion molecule-1 and P25942 mRNA , and numbers of intercellular adhesion molecule-1- or P25942 -positive cells in the grafts were lower in KO mice at 7 and 14 days after transplantation . These reductions resulted in a significant attenuation of P08575 -positive leukocytes adhering to the graft vessel wall in KO mice at 14 days after transplantation . CONCLUSION : Diminished platelet function by Q9H244 receptor deficiency attenuates initiation and strongly inhibits progression of transplant arteriosclerosis in mice by diminishing adhesion molecule expression and leukocyte accumulation in the grafts during the early phase after transplantation . Dependence on phosphoinositide 3-kinase and DB01367 -RAF pathways drive the activity of RAF265 , a novel RAF/ P35968 inhibitor , and RAD001 ( DB01590 ) in combination . Activation of phosphatidylinositol-3-kinase ( PI3K ) -AKT and Kirsten rat sarcoma viral oncogene homologue ( P01116 ) can induce cellular immortalization , proliferation , and resistance to anticancer therapeutics such as epidermal growth factor receptor inhibitors or chemotherapy . This study assessed the consequences of inhibiting these two pathways in tumor cells with activation of P01116 , PI3K-AKT , or both . We investigated whether the combination of a novel RAF/vascular endothelial growth factor receptor inhibitor , RAF265 , with a mammalian target of rapamycin ( P42345 ) inhibitor , RAD001 ( everolimus ) , could lead to enhanced antitumoral effects in vitro and in vivo . To address this question , we used cell lines with different status regarding P01116 , P42336 , and P15056 mutations , using immunoblotting to evaluate the inhibitors , and MTT and clonogenic assays for effects on cell viability and proliferation . Subcutaneous xenografts were used to assess the activity of the combination in vivo . RAD001 inhibited P42345 downstream signaling in all cell lines , whereas RAF265 inhibited RAF downstream signaling only in P15056 mutant cells . In vitro , addition of RAF265 to RAD001 led to decreased AKT , S6 , and P06730 binding protein 1 phosphorylation in HCT116 cells . In vitro and in vivo , RAD001 addition enhanced the antitumoral effect of RAF265 in HCT116 and H460 cells ( both P01116 mut , P42336 mut ) ; in contrast , the combination of RAF265 and RAD001 yielded no additional activity in A549 and MDAMB231 cells . The combination of RAF and P42345 inhibitors is effective for enhancing antitumoral effects in cells with deregulation of both DB01367 -RAF and PI3K , possibly through the cross-inhibition of 4E binding protein 1 and S6 protein . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Human epidermal Langerhans ' cells are targets for the immunosuppressive macrolide tacrolimus ( FK506 ) . BACKGROUND : The immunosuppressive macrolide tacrolimus ( FK506 ) has been shown to inhibit allergic contact dermatitis in animal models as well as in human beings . More recently , successful treatment of atopic dermatitis with an ointment containing tacrolimus has been reported . OBJECTIVES : We explored the effects of this compound on epidermal Langerhans ' cells ( LCs ) , which are known to play an important pathophysiologic role in inflammatory skin diseases . METHODS : The expression of the intracellular FK506 binding protein ( P62942 ) was monitored on freshly isolated and cultured epidermal LCs . Phenotyping and functional exploration of LCs treated with different concentrations of tacrolimus and beta-methasone valerate ( betaMv ) were performed . RESULTS : P62942 is expressed in freshly isolated LCs but is lost while they are maturating into mature dendritic cells . DB00864 inhibited the expression of IL-2R ( CD25 ) and of the costimulatory molecules P33681 ( P33681 .1 ) and P25942 . Expression of MHC class I and II was also affected , whereas P42081 ( P33681 .2 ) expression was not altered . In contrast , betaMv strongly increased the expression of CD25 . Paradoxically , while decreasing P25942 and MHC class I expression , betaMv significantly increased the expression of MHC class II , P33681 , and P42081 on cultured LCs but impaired their allostimulatory activity . DB00864 was about 100 times more potent than betaMv at inhibiting LC stimulatory function . CONCLUSION : DB00864 can exert immunopharmacologic alterations on LCs , which may account , at least in part , for the therapeutic effect of this compound in eczematous skin diseases . DB00644 agonists reduce the migratory and the invasive behavior of androgen-independent prostate cancer cells by interfering with the activity of P05019 . Androgen-independent prostate carcinoma is characterized by a high proliferation rate and by a strong metastatic behavior . We have previously shown that DB00644 agonists exert a direct and specific inhibitory action on the proliferation of androgen-independent prostate cancer cells ( DU 145 ) . These compounds mainly act by interfering with the mitogenic activity of growth factors , such as the insulin-like growth factor-I ( P05019 ) . The present experiments were performed to clarify whether DB00644 agonists might also affect the migratory and the invasive behavior of androgen-independent prostate cancer cells and to define their mechanism of action . First we showed that the DB00644 agonist DB00007 reduces the migration of DU 145 cells towards a chemoattractant and their ability to invade a reconstituted basement membrane . Experiments were then performed to clarify whether the DB00644 agonist might act by interfering with the pro-metastatic activity of P05019 . We found that , in androgen-independent prostate cancer cells , DB00007 : a ) interferes with the P05019 system ( receptor protein expression and tyrosine-phosphorylation ) ; b ) abrogates the P05019 -induced phosphorylation of Akt ( a kinase previously shown by us to mediate the pro-metastatic activity of P05019 in prostate cancer cells ) ; c ) counteracts the migration and the invasive activity of the cells stimulated by P05019 ; d ) abolishes the effects of P05019 on cell morphology , on actin cytoskeleton organization and on alphavbeta3 integrin expression/cellular localization . These data indicate that DB00644 agonists , in addition to their well known antiproliferative effect , can also exert a significant inhibitory activity on the migratory and invasive behavior of androgen-independent prostate cancer cells , expressing the P30968 . DB00644 agonists act by interfering with the pro-metastatic activity of the growth factor P05019 . Platelet Q9H244 receptor inhibition by thienopyridines : status and future . Thienopyridines have a well-established role in the treatment of coronary artery disease , especially in the setting of acute coronary syndromes and percutaneous coronary interventions . DB00208 , the first FDA-approved thienopyridine , was shown to be effective in reducing coronary events in high risk patients , but the original enthusiasm was hampered by concerns about its serious bone marrow toxicity . DB00758 a second generation thienopyridine with lesser side effects , is not only at least as effective as ticlopidine , but in combination with a low dose of aspirin , has been demonstrated to reduce the risk of major cardiovascular events in acute coronary syndrome patients in large-scale , randomised trials . Recent studies have highlighted major flaws in clopidogrel pharmacokinetics due to its delayed onset of action , and much attention has been devoted to the phenomenon of clopidogrel ' resistance ' . Among the novel , third generation thienopyridines , prasugrel as compared to clopidogrel has demonstrated lower inter-patient response variability and a reduced incidence of ischaemic events , but at an increased risk of major bleeding . Currently , several studies are continuing to test new direct Q9H244 receptor antagonists , such as cangrelor and AZD6140 , characterised by a faster reversal of platelet inhibition . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . The thienopyridine derivatives ( platelet adenosine diphosphate receptor antagonists ) , pharmacology and clinical developments . The thienopyridines , ticlopidine and clopidogrel , are antiplatelet drugs . They are prodrugs and are metabolised in the liver to active metabolites that are non-competitive antagonists of the platelet adenosine diphosphate receptor , Q9H244 . Inhibition of platelet aggregation by these drugs is delayed until 24-48 h after administration , with maximal inhibition achieved after 3-5 days . Recovery of platelet function after drug withdrawal is slow ( 7-14 days ) . DB00208 and clopidogrel are effective in preventing atherothrombotic events in cardiovascular , cerebrovascular and peripheral vascular disease . Gastrointestinal side effects and skin rashes are common . However , neutropenia and thrombotic thrombocytopenic purpura are significant and sometimes fatal adverse effects of ticlopidine . DB00758 appears to offer several advantages over ticlopidine : a more rapid onset of action and a lower incidence of neutropenia and thrombotic thrombocytopenic purpura.A combination of clopidogrel and aspirin has become standard for antithrombotic therapy in cardiovascular disease . The anaesthetic considerations of patients taking the thienopyridine compounds are discussed . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . ADP receptors -- targets for developing antithrombotic agents . Platelet P2 receptors -- P47900 , Q9H244 , and P51575 -- constitute the means by which adenine nucleotides can activate platelets . Coactivation of the Galphaq-coupled P47900 and Galphai2-coupled Q9H244 receptors is necessary for ADP-mediated platelet activation , which forms the basis of using P2 antagonists as antithrombotic drugs . P47900 receptor antagonists inhibit platelet activation , while P47900 knockout mice show longer bleeding times than normal mice but few other problems ; however , its ubiquitous expression in other tissues renders P47900 questionable as an antithrombotic target . The Q9H244 receptor is expressed nearly exclusively in platelets and brain , making it an attractive antithrombotic target . Antagonists for the Q9H244 receptor have been developed that either require metabolic activation to covalently inhibit Q9H244 and are irreversible , or simply are competitive in nature and thus reversible . DB00208 and clopidogrel are irreversible Q9H244 antagonists and have been repeatedly proven as clinical antithrombotic agents . In addition , a recently reported Q9H244 antagonist , CS-747 , shows promise as a future antithrombotic drug . The AR-C series of compounds represent reversible Q9H244 antagonists and have been used extensively to characterize the function of Q9H244 in platelets . Clinical studies show that AR-C69931MX is as effective as clopidogrel ; furthermore , the combination of AR-C69931MX ( cangrelor ) and clopidogrel confers greater antagonism of Q9H244 than either antagonist alone . The P51575 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change , and its inhibition or loss has little if any effect on hemostasis . A combination of P47900 and Q9H244 antagonists may represent an additional course of antithrombotic treatment . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies .

MH_train_1025

MH_train_1025

MH_train_1025

interacts_with DB00193?

multiple_choice

Metalloproteinases control brain inflammation induced by pertussis toxin in mice overexpressing the chemokine P13500 in the central nervous system . Inflammatory leukocytes infiltrate the CNS parenchyma in neuroinflammation . This involves cellular migration across various structures associated with the blood-brain barrier : the vascular endothelium , the glia limitans , and the perivascular space between them . Leukocytes accumulate spontaneously in the perivascular space in brains of transgenic ( Tg ) mice that overexpress P13500 under control of a CNS-specific promoter . The Tg mice show no clinical symptoms , even though leukocytes have crossed the endothelial basement membrane . Pertussis toxin ( PTx ) given i.p. induced encephalopathy and weight loss in Tg mice . We used flow cytometry , ultra-small superparamagnetic iron oxide-enhanced magnetic resonance imaging , and immunofluorescent staining to show that encephalopathy involved leukocyte migration across the glia limitans into the brain parenchyma , identifying this as the critical step in inducing clinical symptoms . Metalloproteinase ( MPs ) enzymes are implicated in leukocyte infiltration in neuroinflammation . Unmanipulated Tg mice had elevated expression of tissue inhibitor of metalloproteinase-1 , matrix metalloproteinase ( MMP ) -10 , and -12 mRNA in the brain . PTx further induced expression of tissue inhibitor of metalloproteinase-1 , metalloproteinase disintegrins-12 , P22894 , and -10 in brains of Tg mice . Levels of the microglial-associated MP P51511 were not affected in control or PTx-treated Tg mice . PTx also up-regulated expression of proinflammatory cytokines IL-1beta and P01375 mRNA in Tg CNS . Weight loss and parenchymal infiltration , but not perivascular accumulation , were significantly inhibited by the broad-spectrum MP inhibitor BB-94/ DB03880 . Our finding that MPs mediate PTx-induced parenchymal infiltration to the chemokine-overexpressing CNS has relevance for the pathogenesis of human diseases involving CNS inflammation , such as multiple sclerosis . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . P35372 -dependent and independent components in effects of tramadol . DB00193 is thought to induce analgesia via both opioid and non-opioid pathways , although the precise mechanisms remain to be elucidated . In this study , we investigated the roles of the mu-opioid receptor ( MOP ) in analgesic and rewarding effects of tramadol by using MOP knockout ( KO ) mice . DB00193 -induced antinociception , assessed by hot-plate and tail-flick tests , was significantly reduced in heterozygous and homozygous MOP-KO mice when compared with that in wild-type mice . Interestingly , however , tramadol retained its ability to induce significant antinociception in homozygous MOP-KO mice . The tramadol-induced antinociception remaining in homozygous MOP-KO mice was not significantly affected by methysergide , a serotonin receptor antagonist , but was partially blocked by yohimbine , an adrenaline alpha2 receptor antagonist , and both naloxone , a non-selective opioid receptor antagonist , and yohimbine . In addition , antinociceptive effects of an active tramadol metabolite M1 were abolished or remarkably reduced in MOP-KO mice . On the other hand , neither wild-type nor homozygous MOP-KO mice showed significant place preference for tramadol in a conditioned place preference test , although there were slight tendencies toward preference in wild-type mice and avoidance in homozygous MOP-KO mice . These results strongly support the idea suggested in the previous pharmacological studies that MOP and the adrenaline alpha2 receptor mediate most of the analgesic properties of tramadol . Activation of the P08908 receptor subtype increases rat plasma DB01285 concentration . Serotonergic ( 5-HT ) neuronal pathways regulate the release of adrenocorticotropin hormone ( DB01285 ) from the pituitary gland probably through the action of hypothalamic corticotropin-releasing hormone ( P06850 ) . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a selective P08908 receptor agonist , dose dependently ( 0.016-3 mg/kg s.c. ) increased rat plasma DB01285 concentration . This response was blocked stereoselectively by (-)-pindolol , known to have 5-HT1 antagonist properties , but not by (+)-pindolol , beta 1- , beta 2- or alpha 1-adrenoceptor , dopamine , muscarinic , 5-HT2 or 5- Q9H205 receptor antagonists . Similar increases of plasma DB01285 were induced by other P08908 receptor ligands ( buspirone , ipsapirone and gepirone ) . These results suggest that activation of the P08908 receptor induces the secretion of DB01285 from the rat pituitary gland . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . A vitamin A deficient diet enhances proinflammatory cytokine , P35372 , and HIV-1 expression in the HIV-1 transgenic rat . The HIV-1 ( HIV ) transgenic ( Tg ) rat develops several immune abnormalities in association with clinical impairments that are similar to what are seen with HIV infection in humans . In HIV infection , retinoids and opioids can have separate and potentially combined effects on the clinical course of HIV disease . In these studies , the effects of a vitamin A deficient diet on T cell proinflammatory cytokine and mu opioid receptor ( MOR ) expression were examined in the Tg and in wild-type ( WT ) rats . The effects of the diet on HIV gene expression were also analyzed in the Tg rats . Phytohemagglutinin-stimulated T cells from WT rats on the vitamin A diet and from Tg rats on either diet were more likely to either produce increased percentages of T cells expressing intracytoplasmic P01579 , secrete higher levels of P01375 , and express higher levels of MOR mRNA and surface MOR . Mitogen stimulation also increased Tg rat HIV env , tat , and nef mRNA expression with even higher env and nef mRNA produced in association with the vitamin A deficient diet . All together , these data suggest that a vitamin A deficient diet can result in cellular effects that increase T cell proinflammatory responses and HIV expression , which may alter the course of disease in the HIV Tg rat model . Role of serotonin in the regulation of interferon-gamma production by human natural killer cells . Monocytes , recovered directly from peripheral blood by counter-current centrifugal elutriation ( CCE ) , were shown to provide two regulatory signals for induction of interferon-gamma ( P01579 ) in natural killer ( NK ) cells in response to interleukin-2 ( P60568 ) : an upregulating signal and an inhibitory signal . The inhibitory signal was time-dependent , irreversible , and operating on a pretranslational level , as indicated by the inability of enriched NK cells to accumulate P01579 mRNA in the presence of elutriated monocytes . Monocyte-induced inhibition of P01579 production was abrogated by the biogenic amine serotonin , acting via the 5-hydroxytryptamine , or serotonin ( P08908 ) , subset of serotonin receptors ( 5-HTR ) . Thereby , serotonin effectively promoted P01579 production in the presence of monocytes . We conclude that serotonergic P08908 receptors transduce signals that are required for NK cells to produce P01579 in response to P60568 . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . A 3-D model for P08908 -receptor agonists based on stereoselective methyl-substituted and conformationally restricted analogues of 8-hydroxy-2-(dipropylamino)tetralin . The enantiomers of cis- and trans-1,2,3,4,4a,5,10,10a-octahydro-9-hydroxy-1- propylbenzo[g]quinolines ( 10 and 11 , respectively ) and the enantiomers of trans-1,2,3,4,4a,5,6,10b-octahydro-10- hydroxy-4-propylbenzo[f]quinoline ( 12 ) have been synthesized and their stereochemical and conformational characteristics have been studied by use of X-ray crystallography and molecular mechanics ( P08253 ) calculations . The compounds , which are conformationally restricted analogues of the potent 5-hydroxytryptamine ( 5-HT ) receptor agonist 8-hydroxy-2- (dipropylamino)tetralin ( 8-OH-DPAT ; 1 ) have been evaluated for central 5-HT and dopamine receptor stimulating activity by use of biochemical and behavioral tests in rats . In addition , we have evaluated the ability of these compounds and a number of previously reported analogues to displace [ 3H ] -8-OH-DPAT from P08908 -binding sites . The enantiomers of 12 behave as potent P08908 -receptor agonists , whereas the octahydrobenzo[g]quinoline derivatives are much less potent or inactive . In general , the affinities of the compounds correlate well with their agonist potencies . The set of compounds under study is accommodated by a novel computer-graphics-derived model for P08908 -receptor agonism . The model consists of a flexible pharmacophore and a partial receptor-excluded volume . Synergistic proapoptotic effects of the two tyrosine kinase inhibitors pazopanib and lapatinib on multiple carcinoma cell lines . DB06589 and lapatinib are two tyrosine kinase inhibitors that have been designed to inhibit the P15692 tyrosine kinase receptors 1 , 2 and 3 ( pazopanib ) , and the P00533 and P04626 receptors in a dual manner ( lapatinib ) . DB06589 has also been reported to mediate inhibitory effect on a selected panel of additional tyrosine kinases such as P09619 and c-kit . Here , we report that pazopanib and lapatinib act synergistically to induce apoptosis of A549 non-small-cell lung cancer cells . Systematic assessment of the kinome revealed that both pazopanib and lapatinib inhibited dozens of different tyrosine kinases and that their combination could suppress the activity of some tyrosine kinases ( such as c- DB00134 ) that were not or only partially affected by either of the two agents alone . We also found that pazopanib and lapatinib induced selective changes in the transcriptome of A549 cells , some of which were specific for the combination of both agents . Analysis of a panel of unrelated human carcinoma cell lines revealed a signature of 52 genes whose up- or downregulation reflected the combined action of pazopanib and lapatinib . Indeed , pazopanib and lapatinib exerted synergistic cytotoxic effects on several distinct non-small-cell lung cancer cells as well as on unrelated carcinomas . Altogether , these results support the contention that combinations of tyrosine kinase inhibitors should be evaluated for synergistic antitumor effects . Such combinations may lead to a ' collapse ' of pro-survival signal transduction pathways that leads to apoptotic cell death . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . Clinical pharmacology of serotonin-altering medications for decreasing alcohol consumption . Variations in serotonin neurotransmission influence alcohol consumption ( AC ) . Levels of 5-HT and metabolites are low in some brain regions of alcohol preferring rats and in P04141 of alcoholics . Pharmacological treatments which enhance serotonergic neurotransmission ( uptake inhibitors , releasers , agonists ) consistently reduce AC in rats . Serotonin uptake inhibitors ( SUI ; e.g. , citalopram , fluoxetine ) have been studied extensively in humans . In several double-blind randomized , placebo-controlled clinical trials , SUI have consistently decreased AC by averages of 15 % to 20 % in nondepressed mildly/moderately dependent alcoholics who received no other treatment . Effects were dose-dependent and not related to side effects ( few and mild ) or changes in anxiety or depression ( not observed ) . SUI also decreased desire to drink and liking for alcohol , thus suggesting a mechanism for effects . Other drugs acting on the 5-HT system have been tested in humans , but results are difficult to interpret . For example , buspirone , a P08908 receptor partial agonist , reduced anxiety and alcohol craving , but not AC ; a 5-HT partial agonist , m-CPP , increased alcohol craving in abstinent alcoholics ; modest reductions in AC were observed with a 5- Q9H205 antagonist , ondansetron ( 0.5 mg/day , but not 4 mg/day ) . The therapeutic potentials of these medications are being studied . For example , SUI effects on AC were enhanced by a brief psychosocial intervention . Since SUI decrease urge to drink , they may be suitable pharmacological adjuncts in relapse prevention strategies . SUI and other serotonin-altering medications are promising new neuropharmacological treatments for reducing AC . P18509 , interleukin-6 and glucocorticoids regulate the release of vascular endothelial growth factor in pituitary folliculostellate cells . There is increasing evidence that hormones play an important role in the control of endothelial cell function and growth by regulating the production of vascular endothelial growth factor ( P15692 ) . P15692 regulates vascular permeability and represents the most powerful growth factor for endothelial cells . In the normal anterior pituitary , P15692 has been detected only in folliculostellate ( FS ) cells . In the present study , the regulation of the release of P15692 from FS-like mouse TtT/GF cells , and from FS cells of rat pituitary monolayer cell cultures was investigated using a specific P15692 ELISA . Basal release of P15692 was demonstrated in cultures of both TtT/GF cells and rat pituitary cells . Interestingly , the P15692 secretion was stimulated by both forms of pituitary adenylate cyclase-activating polypeptide ( PACAP-38 and PACAP-27 ) , indicating that this hypothalamic peptide regulates endothelial cell function and growth within the pituitary . P15692 secretion was also stimulated by interleukin-6 ( P05231 ) whereas basal , P05231 - and PACAP-stimulated secretion was inhibited by the synthetic glucocorticoid dexamethasone . The inhibitory action of dexamethasone was reversed by the glucocorticoid receptor antagonist DB00834 , suggesting that in FS cells functional glucocorticoid receptors mediate the inhibitory action of glucocorticoids on the P15692 secretion . The endocrine and auto-/paracrine control of P15692 production in pituitary FS cells by PACAP , P05231 and glucocorticoids may play an important role both in angiogenesis and vascular permeability regulation within the pituitary under physiological and pathophysiological conditions . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Central mechanisms of vomiting . Nausea and vomiting ( emesis ) occur under a variety of conditions in response to activation of one or more emetic triggers . The act of vomiting is coordinated by neuronal circuitry located in the brain stem between the obex and the retrofacial nucleus , including the region extending from the nucleus of the solitary tract through the lateral tegmental field of the reticular formation to the ventrolateral medulla . The area postrema , medullary midline , and certain higher brain centers are also important for vomiting . The sensation of nausea is thought to involve the cerebral cortex . The most effective near-term treatment for combating nausea and vomiting associated with cyclic vomiting syndrome may come from experimental drugs ( P25103 antagonists , P08908 receptor agonists ) or P6 acustimulation , which have been shown to combat nausea and vomiting in response to a broad spectrum of emetic challenges and thus presumably act on central emetic mechanisms . P35372 and P00533 contribute to skin pigmentation differences between Indigenous Americans and Europeans . Contemporary variation in skin pigmentation is the result of hundreds of thousands years of human evolution in new and changing environments . Previous studies have identified several genes involved in skin pigmentation differences among African , Asian , and European populations . However , none have examined skin pigmentation variation among Indigenous American populations , creating a critical gap in our understanding of skin pigmentation variation . This study investigates signatures of selection at 76 pigmentation candidate genes that may contribute to skin pigmentation differences between Indigenous Americans and Europeans . Analysis was performed on two samples of Indigenous Americans genotyped on genome-wide SNP arrays . Using four tests for natural selection -- locus-specific branch length ( LSBL ) , ratio of heterozygosities ( lnRH ) , Tajima 's D difference , and extended haplotype homozygosity ( EHH ) -- we identified 14 selection-nominated candidate genes ( SNCGs ) . SNPs in each of the SNCGs were tested for association with skin pigmentation in 515 admixed Indigenous American and European individuals from regions of the Americas with high ground-level ultraviolet radiation . In addition to Q71RS6 and Q9UMX9 , genes previously associated with European/non-European differences in skin pigmentation , P35372 and P00533 were associated with variation in skin pigmentation in New World populations for the first time . Identification of a variant in P35968 associated with serum P35968 and pharmacodynamics of DB06589 . PURPOSE : P15692 receptor ( VEGFR ) kinases are important drug targets in oncology that affect function of systemic endothelial cells . To discover genetic markers that affect VEGFR inhibitor pharmacodynamics , we performed a genome-wide association study of serum soluble vascular P35968 concentrations [ sVEGFR2 ] , a pharmacodynamic biomarker for P35968 inhibitors . EXPERIMENTAL DESIGN : We conducted a genome-wide association study ( GWAS ) of [ sVEGFR2 ] in 736 healthy Old Order Amish volunteers . Gene variants identified from the GWAS were genotyped serially in a cohort of 128 patients with advanced solid tumor with baseline [ sVEGFR2 ] measurements , and in 121 patients with renal carcinoma with [ sVEGFR2 ] measured before and during pazopanib therapy . RESULTS : rs34231037 ( C482R ) in P35968 , the gene encoding sVEGFR2 was found to be highly associated with [ sVEGFR2 ] , explaining 23 % of the variance ( P = 2.7 × 10(-37) ) . Association of rs34231037 with [ sVEGFR2 ] was replicated in 128 patients with cancer with comparable effect size ( P = 0.025 ) . Furthermore , rs34231037 was a significant predictor of changes in [ sVEGFR2 ] in response to pazopanib ( P = 0.01 ) . CONCLUSION : Our findings suggest that genome-wide analysis of phenotypes in healthy populations can expedite identification of candidate pharmacogenetic markers . Genotyping for germline variants in P35968 may have clinical utility in identifying patients with cancer with unusual sensitivity to effects of P35968 kinase inhibitors . Regulation of P14061 and P18405 in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta-HSD and 5alpha-reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta-HSD and 5alpha-reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) -4 , and P05231 . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 ( encoding 17beta-HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 ( encoding 5alpha-reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10(-9) to 10(-7) M androstenedione or estradiol increased expression of P14061 , while testosterone decreased the expression of this gene . P18405 expression was increased in the presence of 5alpha- DB02901 although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6-hydroxypregnanolone , produced no effects on expression of either P14061 or P18405 . Treatment with 0.1-10 ng/ml of P05112 or P05231 also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 -protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 . Furthermore , it appears that lymphocyte P14061 and P18405 are regulated to some extent by specific steroids . Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons . 1. The basolateral amygdala ( P00519 ) nuclei contribute to the process of anxiety . GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release . In mechanically dissociated rat P00519 neurons , spontaneous miniature inhibitory postsynaptic currents ( mIPSCs ) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation . 2 . 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude . The serotonergic effect was mimicked by the P08908 specific agonist 8-OH DPAT ( 8-hydroxy-2-(di-n-propylamino)tetralin ) and blocked by the P08908 antagonist spiperone . 3 . The GTP-binding protein inhibitor N-ethylmaleimide removed the serotonergic inhibition of mIPSC frequency . In either K+-free or Ca2+-free external solution , 5-HT could inhibit mIPSC frequency . 4 . High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+ . 5 . DB02587 , an activator of adenylyl cyclase ( AC ) , significantly increased synaptic GABA release frequency . Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution . 6 . Ruthenium Red ( RR ) , an agent facilitating the secretory process in a Ca2+-independent manner , increased synaptic GABA release . 5-HT also suppressed RR-facilitated mIPSC frequency . 7 . We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC- DB02527 signal transduction pathway via a G-protein-coupled P08908 receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals . Neurogenetics of aggressive behavior : studies in primates . Aggressive behavior can have adaptive value in certain environmental contexts , but when extreme or executed inappropriately , can also lead to maladaptive outcomes . Neurogenetic studies performed in nonhuman primates have shown that genetic variation that impacts reward sensitivity , impulsivity , and anxiety can contribute to individual differences in aggressive behavior . Genetic polymorphisms in the coding or promoter regions of the Mu-Opioid Receptor ( P35372 ) , DB01285 Releasing Hormone ( P06850 ) , Monoamine Oxidase A ( P21397 ) , Dopamine D4 Receptor ( P21917 ) , and Serotonin Transporter ( P31645 ) genes have been shown to be functionally similar in humans and rhesus macaques and have been demonstrated to contribute to individual differences in aggression . This body of literature suggests mechanisms by which genetic variation that promotes aggressivity could simultaneously increase evolutionary success while making modern humans more vulnerable to psychopathology . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks .

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Q9GZX6 /IL-22R1 axis and P05109 /A9 alarmins in human osteoarthritic and rheumatoid arthritis synovial fibroblasts . OBJECTIVES : Fibroblast-like synoviocytes ( FLSs ) are crucial players in the pathogenesis of synovitis in rheumatic diseases . Targeting FLS activation represents an approach to the development of therapeutic strategies . Our aim was to investigate whether the microenvironment of inflamed joints could modulate the expression of Q9GZX6 and IL-22R1 on OA and RA FLSs . We also examined the effect of Q9GZX6 on FLS activation as well as on their Q16552 -related responses . METHODS : Q9GZX6 and IL-22R1 expression was studied by RT-PCR and immunoblotting . Proliferation was measured by an ELISA kit . Q16552 receptors , p19IL-23 and alarmins were analysed by RT-PCR . Q16552 receptor expression was evaluated by flow cytometry . P03956 and IL-23 were measured by ELISA . P05109 /A9 expression was detected by immunofluorescence and ELISA . P40763 ( P40763 ) phosphorylation was quantified using a cell-based ELISA kit . RESULTS : Q9GZX6 and IL-22R1 were expressed constitutively in FLSs . We demonstrated that P05109 and P06702 were synthesized in FLSs . We reported that inflammatory mediators increased the expression of the Q9GZX6 /IL-22R1 axis , amplifying FLS activation . Q9GZX6 enhanced FLS proliferation and up-regulated P03956 and P05109 /A9 production . P40763 phosphorylation was induced after Q9GZX6 treatment and the stimulatory effect of Q9GZX6 on P05109 /A9 was reduced after the activities of O60674 ( O60674 ) and P52333 were blocked . We showed an inhibitory action of Q9GZX6 on IL-23 and Q8NAC3 expression in RA FLSs and on Q96F46 in OA FLSs . CONCLUSION : Therapies based on the pharmacological disruption signalling of Q9GZX6 could be beneficial for the treatment of rheumatic diseases . The restricted expression of IL-22R1 to non-lymphoid cells could lead to a reduction of side effects mediated by immune responses . Selective JAK/ P40763 signalling regulates transcription of colony stimulating factor-2 and -3 in Concanavalin-A-activated mesenchymal stromal cells . Human bone marrow-derived mesenchymal stromal cells ( MSCs ) express Toll-like receptors ( TLRs ) and produce cytokines and chemokines , all of which contribute to these cells ' immunomodulatory and proangiogenic properties . Among the secreted cytokines , colony-stimulating factors ( CSFs ) regulate angiogenesis through activation of endothelial cell proliferation and migration . Since O60682 are recruited within hypoxic tumors where they signal paracrine-regulated angiogenesis , the aim of this study was to evaluate which P04141 members are expressed and are inducible in activated O60682 . Furthermore , we investigated the JAK/ P35610 signal transducing pathway that may impact on P04141 transcription . O60682 were activated with Concanavalin-A ( ConA ) , a TLR-2/6 agonist as well as a membrane type-1 matrix metalloproteinase ( P50281 ) inducer , and we found increased transcription of granulocyte macrophage- P04141 ( GM- P04141 , P04141 -2 ) , granulocyte P04141 ( DB00099 , P04141 -3 ) , and P50281 . Gene silencing of either P40763 or P50281 prevented ConA-induced phosphorylation of P40763 , and reversed ConA effects on P04141 -2 and P04141 -3 . Treatment with the Janus Kinase (JAK)2 inhibitor AG490 antagonized the ConA induction of P50281 and P04141 -2 , while the pan-JAK inhibitor DB08895 reversed ConA-induced P04141 -2 and -3 gene expression . Silencing of O60674 prevented the ConA-mediated increase of P04141 -2 , while silencing of P23458 , P52333 and P29597 prevented the increase in P04141 -3 . Given that combined TLR-activation and locally-produced P04141 -2 and P04141 -3 could regulate immunomodulation and neovascularization , pharmacological targeting of TLR-2/6-induced P50281 /JAK/ P40763 signalling pathway may prevent O60682 contribution to tumor development . DB00877 protects against myocardial ischemia-reperfusion injury through O60674 - P40763 signaling pathway . DB00877 ( Sirolimus® ) is used to prevent rejection of transplanted organs and coronary restenosis . We reported that rapamycin induced cardioprotection against ischemia-reperfusion ( I/R ) injury through opening of mitochondrial K( DB00171 ) channels . However , signaling mechanisms in rapamycin-induced cardioprotection are currently unknown . Considering that P40763 is protective in the heart , we investigated the potential role of this transcription factor in rapamycin-induced protection against ( I/R ) injury . Adult male ICR mice were treated with rapamycin ( 0.25mg/kg , i.p. ) or vehicle ( DB01093 ) with/without inhibitor of O60674 ( AG-490 ) or P40763 ( stattic ) . One hour later , the hearts were subjected to I/R either in Langendorff mode or in situ ligation of left coronary artery . Additionally , primary murine cardiomyocytes were subjected to simulated ischemia-reoxygenation ( SI/RO ) injury in vitro . For in situ targeted knockdown of P40763 , lentiviral vector containing short hairpin RNA was injected into the left ventricle 3 weeks prior to initiating I/R injury . Infarct size , cardiac function , and cardiomyocyte necrosis and apoptosis were assessed . DB00877 reduced infarct size , improved cardiac function following I/R , and limited cardiomyocyte necrosis as well as apoptosis following SI/RO which were blocked by AG-490 and stattic . In situ knock-down of P40763 attenuated rapamycin-induced protection against I/R injury . DB00877 triggered unique cardioprotective signaling including phosphorylation of P29323 , P40763 , P29474 and glycogen synthase kinase-3ß in concert with increased prosurvival Bcl-2 to Bax ratio . Our data suggest that O60674 - P40763 signaling plays an essential role in rapamycin-induced cardioprotection . We propose that rapamycin is a novel and clinically relevant pharmacological strategy to target P40763 activation for treatment of myocardial infarction . Cyclophosphamide and other new agents for the treatment of severe aplastic anemia . Severe aplastic anemia ( P0DJI8 ) has a poor prognosis in the absence of treatment . Current accepted therapeutic strategies include allogeneic stem-cell transplantation and immunosuppression , both resulting in long-term survival in the majority of patients . Although human leukocyte antigen ( HLA ) -matched sibling stem-cell transplantation is highly effective , the 25 % probability of finding a suitable sibling donor within a family renders this approach available to only a minority of patients . Transplantation using HLA-matched , unrelated donors carries a high risk of treatment failure along with considerable toxicity . While combined immunosuppression with both antithymocyte globulin ( ATG ) and cyclosporine A ( Q13216 ) produces hematologic improvement in most patients , relapse is common . Late evolution of aplastic anemia to other serious hematologic disorders , including paroxysmal nocturnal hemoglobinuria ( PNH ) , myelodysplasia , and acute leukemia , is also a significant problem following treatment with ATG/ Q13216 . Recently , results of immunosuppression in P0DJI8 with another potent immunosuppressive agent , cyclophosphamide , were reported in a small number of patients . The overall response rate was similar to that seen with ATG/ Q13216 , but relapse and late clonal disease were not observed during a long period of follow-up . A larger randomized trial comparing sustained hematologic response rates to either conventional immunosuppression with ATG/ Q13216 or high-dose cyclophosphamide and Q13216 is now underway ; secondary end points include response duration , event-free survival , and overall survival . Additionally , a number of protocols designed to test the efficacy of alternative immunosuppressive or immunomodulatory agents are being developed . Influence of Janus kinase inhibition on interleukin 6-mediated induction of acute-phase serum amyloid A in rheumatoid synovium . OBJECTIVE : Inhibition of intracellular signal transduction is considered to be a therapeutic target for chronic inflammation . The new Janus kinase (JAK)3 inhibitor CP690,550 has shown efficacy in the treatment of rheumatoid arthritis ( RA ) . We investigated the influence of JAK/ P35610 inhibition using CP690,550 on the induction of acute-phase serum amyloid A ( P0DJI8 ) , which is triggered by interleukin 6 ( P05231 ) stimulation in rheumatoid fibroblast-like synoviocytes ( RA-FLS ) . METHODS : P05231 -stimulated gene expression of the acute-phase serum amyloid A genes ( A- P0DJI8 ; encoded by P0DJI8 + P0DJI9 ) and P35542 was analyzed by reverse transcriptase-polymerase chain reaction . The intracellular signaling pathway mediating the effects of CP690,550 on P05231 -stimulated JAK/ P35610 activation was assessed by measuring the phosphorylation levels using Western blots . RESULTS : P05231 trans-signaling induced A- P0DJI8 messenger RNA ( mRNA ) expression in RA-FLS . By contrast P05231 stimulation did not affect P35542 mRNA expression , which is expressed constitutively in RA-FLS . P05231 stimulation elicited rapid phosphorylation of O60674 and P40763 , which was blunted by CP690,550 . CP690,550 abrogated P05231 -mediated A- P0DJI8 mRNA expression in RA-FLS . Similarly , CP690,550 inhibited P05231 -mediated A- P0DJI8 mRNA expression in human hepatocytes . CONCLUSION : Our data indicated that CP690,550 blocked P05231 -induced O60674 / P40763 activation , as well as the induction of A- P0DJI8 . Inhibition of P05231 -mediated proinflammatory signaling pathways by CP690,550 may represent a new antiinflammatory therapeutic strategy for RA and AA amyloidosis . The attenuation of experimental lung metastasis by a bile acid acylated-heparin derivative . The inhibitory efficacies of new bile acid acylated-heparin derivative ( heparin-DOCA ) were evaluated on experimental lung metastasis . We evaluated the effect of heparin-DOCA on intercellular interactions including those between B16F10 and thrombin-activated platelets and P01375 -activated HUVECs , and between B16F10 and immobilized mouse P16109 . In addition , the inhibitory effects of heparin-DOCA on adhesion and invasion of B16F10 to Matrigel were studied . In an animal mouse study , the blood clot formation and the retention of red fluorescence protein ( RFP ) -B16F10 in lungs were assessed after heparin-DOCA and RFP-B16F10 intravenous administration . Furthermore , we investigated the anti-metastatic effect of heparin-DOCA against lung metastasis induced by B16F10 and SCC7 . DB01109 -DOCA inhibited intercellular interactions between B16F10 and activated platelets or activated HUVECs by blocking P- and P16581 -mediated interactions . Moreover , it reduced adhesion and invasion of B16F10 to Q13201 , thereby affecting the reduction of early retention of B16F10 in the lung . DB01109 -DOCA attenuated lung colony formation on the surfaces and in interior of the lung , and attenuated metastasis by B16F10 and SCC7 . These results suggest that heparin-DOCA may have potentials as therapeutic agent that prevents tumor metastasis and progression . Physiology and pathophysiology of eryptosis . Suicidal erythrocyte death ( eryptosis ) is characterized by cell shrinkage , cell membrane blebbing , and cell membrane phospholipid scrambling with phosphatidylserine exposure at the cell surface . Eryptotic cells adhere to the vascular wall and are rapidly cleared from circulating blood . Eryptosis is stimulated by an increase in cytosolic Ca(2)+ activity , ceramide , hyperosmotic shock , oxidative stress , energy depletion , hyperthermia , and a wide variety of xenobiotics and endogenous substances . Inhibitors of eryptosis include erythropoietin and nitric oxide . Enhanced eryptosis is observed in diabetes , renal insufficiency , hemolytic uremic syndrome , sepsis , mycoplasma infection , malaria , iron deficiency , sickle cell anemia , beta-thalassemia , glucose-6-phosphate dehydrogenase-( P11413 ) deficiency , hereditary spherocytosis , paroxysmal nocturnal hemoglobinuria , Wilson 's disease , myelodysplastic syndrome , and phosphate depletion . Eryptosis is further enhanced in gene-targeted mice with deficient annexin 7 , cGMP-dependent protein kinase type I ( Q13976 ) , AMP-activated protein kinase ( AMPK ) , anion exchanger 1 ( P02730 ) , adenomatous polyposis coli ( P25054 ) , and Q9UEF7 , as well as in mouse models of sickle cell anemia and thalassemia . Decreased eryptosis is observed in mice with deficient phosphoinositide-dependent kinase 1 ( PDK1 ) , platelet activating factor ( Q15004 ) receptor , transient receptor potential channel 6 ( Q9Y210 ) , janus kinase 3 ( P52333 ) , and taurine transporter ( TAUT ) . Eryptosis may be a useful mechanism to remove defective erythrocytes prior to hemolysis . Excessive eryptosis may , however , compromise microcirculation and lead to anemia . Targeting P52333 in kidney transplantation : current status and future options . PURPOSE OF REVIEW : This review will discuss the mechanism of action and important clinical trial data in renal transplantation for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . RECENT FINDINGS : JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared with vehicle control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new-onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . SUMMARY : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . Genetic analysis of expression profile involved in retinoid metabolism in non-alcoholic fatty liver disease . AIM : The patients with non-alcoholic fatty liver disease ( NAFLD ) have been reported to be at greater risk for progression to chronic liver disease including liver cirrhosis ( LC ) . To examine the mechanisms for the progression of NAFLD , a genetic analysis of hepatic expression profile in retinoid metabolism in NAFLD was performed since the loss of retinoid signaling is associated with the progression of liver disease via reactive oxygen species ( ROS ) generation . METHODS : Fifty-one genes , which are associated with retinoid metabolism and action , were examined in thirty six subjects including 17 patients with simple steatosis , 11 with non-alcoholic steatohepatitis ( NASH ) and eight controls were examined by real-time reverse transcriptase polymerase chain reaction . Immunohistochemical study was also done by 3 kinds of antibodies . RESULTS : Higher expression of P09455 O95237 , DGT1/2 and CES1 in NAFLD suggests that mutual conversion between retinyl ester and retinal occurs actively . Expression of P07327 /2/3 , Q92781 /10/11 , O75911 and RALDH1/3 was increased in NAFLD , suggesting that oxidation process from retinol to all-trans retinoic acid ( DB00755 ) was enhanced . Importantly , greater expression of O43174 indicated that degradation of DB00755 was enhanced in NAFLD . Further , expression of P00441 /2 , catalase , thioredoxin and uncoupling protein 2 was also enhanced . CONCLUSION : Hyperdynamic state of retinoid metabolism is present in the liver tissues with NAFLD , which may be a putative mechanism by which NAFLD progresses to chronic liver disease including LC . Opposing actions of extracellular signal-regulated kinase ( P29323 ) and signal transducer and activator of transcription 3 ( P40763 ) in regulating microtubule stabilization during cardiac hypertrophy . Excessive proliferation and stabilization of the microtubule ( MT ) array in cardiac myocytes can accompany pathological cardiac hypertrophy , but the molecular control of these changes remains poorly characterized . In this study , we examined MT stabilization in two independent murine models of heart failure and revealed increases in the levels of post-translationally modified stable MTs , which were closely associated with P40763 activation . To explore the molecular signaling events contributing to control of the cardiac MT network , we stimulated cardiac myocytes with an α-adrenergic agonist phenylephrine ( PE ) , and observed increased tubulin content without changes in detyrosinated ( glu-tubulin ) stable MTs . In contrast , the hypertrophic interleukin-6 ( P05231 ) family cytokines increased both the glu-tubulin content and glu-MT density . When we examined a role for P29323 in regulating cardiac MTs , we showed that the MEK/ P29323 -inhibitor U0126 increased glu-MT density in either control cardiac myocytes or following exposure to hypertrophic agents . Conversely , expression of an activated Q02750 mutant reduced glu-tubulin levels . Thus , P29323 signaling antagonizes stabilization of the cardiac MT array . In contrast , inhibiting either O60674 with AG490 , or P40763 signaling with Stattic or siRNA knockdown , blocked cytokine-stimulated increases in glu-MT density . Furthermore , the expression of a constitutively active P40763 mutant triggered increased glu-MT density in the absence of hypertrophic stimulation . Thus , P40763 activation contributes substantially to cytokine-stimulated glu-MT changes . Taken together , our results highlight the opposing actions of P40763 and P29323 pathways in the regulation of MT changes associated with cardiac myocyte hypertrophy . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . P52333 inhibition significantly attenuates psoriasiform skin inflammation in P05107 mutant PL/J mice . P52333 , a member of the Janus kinase family , is predominantly expressed in hemopoietic cells and binds specifically to the common gamma chain of a subfamily of cytokine receptors that includes P60568 , P05112 , P13232 , P15248 , P40933 , and Q9HBE4 . Previous studies suggest that this tyrosine kinase plays key roles in mediating T cell functions , and inhibition of P52333 has been shown to prevent graft rejection and decrease the severity of arthritis in rodent models . However , the functions of P52333 in the development of skin immune responses and diseases such as psoriasis have not been determined . P05107 mutant PL/J mice develop spontaneous T cell-dependent psoriasiform skin disease with several similarities to human psoriasis . In this study , we treated mice with established skin disease with R348 , a small molecule inhibitor of P52333 , and observed a marked attenuation of skin lesions following 6 wk of treatment . Histological analyses revealed major reductions of both epidermal and dermal lesion severity scores in R348-treated P05107 -deficient PL/J mice compared with vehicle controls , which was associated with decreased P01730 (+) T cell infiltration . In addition , systemic levels of Q16552 , Q9GZX6 , IL-23 , and P01375 were significantly lower in mice receiving the compound , and T cells isolated from R348-treated mice also showed reduced phosphorylation of Stat5 after stimulation with P60568 . These findings suggest that small-molecule inhibitors of P52333 may be useful in the treatment of inflammatory skin diseases such as psoriasis and strongly implicate JAK signaling events as important in the pathogenesis of this disease . T lymphocytes are targets for platelet- and trophoblast-derived microvesicles during pregnancy . Microvesicles ( MVs ) can derive from several cell types and their membranes contain cell surface elements . Their role is increasingly recognized in cell-to-cell communication , as they act as both paracrine and remote messengers , occurring in circulating form as well as in plasma . Successful pregnancy requires a series of interactions between the maternal immune system and the implanted fetus , such that the semi-allograft will not be rejected . These interactions occur at the materno-placental interface and/or at a systemic level . In the present study we identified for the first time the in vivo plasma pattern of the MVs of third-trimester , healthy pregnant women , their cellular origin , and their target cells using flow cytometry and confocal laser microscopy . We searched for the cellular target molecules of thrombocyte-derived MVs with the help of neutralizing antibodies . We examined the in vitro effects of MVs on P40763 phosphorylation of primary lymphocytes and Jurkat cells . We found that both placental trophoblast-derived and maternal thrombocyte-derived MVs bind to circulating peripheral T lymphocytes , but not to B lymphocytes or NK cells . We were able to show that the P16109 ( CD62P ) - Q14242 ( CD162 ) interaction is one mechanism binding platelet-derived MVs to T cells . We were also able to demonstrate that MV-lymphocyte interactions induce P40763 phosphorylation in T cells . Our findings indicate that both thrombocyte- and trophoblast-derived MVs may play an important role in the immunomodulation of pregnancy . We suggest that the transfer of different signals via MVs represents a novel form of communication between the placenta and the maternal immune system , and that MVs contribute to the establishment of stable immune tolerance to the semi-allograft fetus . P12821 ( P12821 ) and Q9BYF1 levels in the cerebrospinal fluid of patients with multiple sclerosis . BACKGROUND : We reported a reduction in the levels of angiotensin II in cerebrospinal fluid ( P04141 ) from patients with multiple sclerosis ( MS ) . OBJECTIVE AND METHODS : To clarify the mechanism underlying this reduction , we assayed angiotensin-converting enzyme ( P12821 ) and Q9BYF1 concentrations along with angiotensin II concentrations in P04141 samples from 20 patients with MS and 17 controls with non-neurological diseases . RESULTS : P12821 levels were significantly elevated in patients with MS compared with controls ( 48.42 +/- 4.84 vs 44.71 +/- 3.9 pg/mL ) , whereas Q9BYF1 levels were significantly reduced ( 2.56 +/- 0.26 vs 2.78 +/- 0.24 pg/mL ) , acting toward a normalization of angiotensin II levels . CONCLUSION : These results further indicate an alteration of the intrathecal renin-angiotensin system in patients with MS . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Multiple Gi proteins participate in nerve growth factor-induced activation of c-Jun N-terminal kinases in PC12 cells . Nerve growth factor ( P01138 ) -mediated activation of mitogen-activated protein kinases ( MAPK ) is critical for differentiation and apoptosis of PC12 cells . Since P01138 employs stress-activated c-Jun N-terminal kinase ( JNK ) to regulate both programmed cell death and neurite outgrowth of PC12 cells , we examined P01138 -regulated JNK activity and the role of G(i/o) proteins . Induction of JNK phosphorylation by P01138 occurred in a time- and dose-dependent manner and was partially inhibited by pertussis toxin ( PTX ) . To discern the participation of various signaling intermediates , PC12 cells were treated with specific inhibitors prior to P01138 challenge . P01138 -elevated JNK activity was abolished by inhibitors of JNK , p38 MAPK , Src , P52333 and Q02750 /2 . P01138 -dependent JNK phosphorylation became insensitive to PTX treatment upon transient expressions of Galpha(z) or the PTX-resistant mutants of Galpha(i1-3) and Galpha(oA) . Collectively , these studies indicate that P01138 -dependent JNK activity may be mediated via G(i1-3) proteins , P52333 , Src , p38 MAPK and the MEK/ P29323 cascade . Predicting resistance by selection of signaling pathways . P00533 ( P00533 ) mutations occur in 17 % of non-small-cell lung cancer ( NSCLC ) patients with notable response to single agent therapy but with low complete remission rate and , eventually , disease progression . Priming O43521 , a pro-apoptotic signaling Q7L3V2 , induces sensitivity to erlotinib in P00533 -mutant cell lines . Synthetic lethal approaches and preemptive therapies based on the initial expression of O43521 may significantly improve the treatment outcome . P00533 mutations result in transient pro-death imbalance of survival and apoptotic signaling in response to P00533 inhibition . SHP2 is essential to the balance between P29323 and the phosphoinositide-3-kinase ( PI3K ) /AKT and signal transducer activator of transcription ( P35610 ) activity , while P42345 can be an additional marker for patients with high O43521 expression . Furthermore , stromal hepatocyte growth factor ( P14210 ) confers P00533 tyrosine kinase inhibitor ( TKI ) resistance and induces interreceptor crosstalk with integrin-b4 , Eph2 , CUB domain-containing protein-1 ( Q9H5V8 ) , P30530 and P23458 . Only by understanding better , and in more depth , complex cancer molecular biology will we have the information that will help us to design strategies to augment efficacy of P00533 TKIs and offer our patients the best , most correct therapeutic option . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Survivin expression in glioblastomas correlates with proliferation , but not with apoptosis . BACKGROUND : Survivin is expressed in proliferating tissues and in tumors . It is a member of the inhibitory apoptosis protein ( IAP ) family known to regulate mitosis and to inhibit apoptosis . It has therefore been regarded as a target for therapies . In malignant gliomas it increases with malignancy , even though in glioblastomas it does not seem to correlate with outcome . MATERIALS AND METHODS : Survivin was immunohistochemically studied in 39 selected viable glioblastoma areas belonging to 20 cases which were assayed for apoptosis , using a TUNEL assay , caspase-3 , poly(ADP-ribose)polymerase 1 ( P09874 ) , Bid ( P55957 ) and with the proliferation index Ki-67/MIB-1 and mitotic index ( MI ) . RESULTS : A positive linear correlation was found between the survivin labelling index ( LI ) and the Ki-67/MIB-1 LI and MI . No inverse correlation was found with apoptosis . CONCLUSION : This double behavior can be attributed to mechanisms mediating survivin activity , either as a mitosis regulator and apoptosis inhibitor , and should be taken into account in therapeutic strategies using survivin . A pharmacokinetic and clinical assessment of tofacitinib for the treatment of rheumatoid arthritis . INTRODUCTION : Rheumatoid arthritis ( RA ) is a chronic painful and debilitating autoimmune disease . Although the outcome for patients with RA has improved markedly in the past decades , driven largely by the advent of biological disease-modifying antirheumatic drugs ( DMARDs ) and updated management strategies , adequate disease control can not be achieved in a substantial proportion of patients . Since RA is a syndrome with different biological subsets , DMARDs , with a novel mechanism of action , may represent a valuable addition to the current armamentarium . DB08895 is a novel synthetic DMARD that selectively inhibits Janus kinases ( JAKs ) , particularly P23458 and P52333 . AREAS COVERED : This review describes the pharmacokinetics of tofacitinib . Furthermore , the article summarizes and comments the drug 's efficacy and safety profile in RA patients . The authors furthermore assess data derived from the FDA 's RA development program . EXPERT OPINION : DB08895 is an oral synthetic DMARD displaying linear pharmacokinetics . Metabolism , primarily mediated by P08684 , accounts for 70 % of the total clearance of the drug ; the remaining 30 % are renally excreted . DB08895 monotherapy , or in combination with traditional DMARDs , has demonstrated its efficacy while having an acceptable safety profile in RA patients who have responded inadequately to current DMARDs , including P01375 antagonists . In view of its undetermined benefit to risk ratio , in the real-world population , tofacitinib should , for now , only be prescribed to selected patients . The JAK inhibitor , tofacitinib , reduces the T cell stimulatory capacity of human monocyte-derived dendritic cells . OBJECTIVE : DB08895 , which is a Janus kinase ( JAK ) inhibitor , has shown clinical effects in the treatment of rheumatoid arthritis . JAKs are important kinases in lymphocyte differentiation ; however , their function in dendritic cells ( DCs ) is unknown . In this study , the function of JAKs in DCs was investigated with tofacitinib . METHODS : The effects of tofacitinib on the maturation of human monocyte-derived DCs induced by lipopolysaccharide ( LPS ) stimulation were investigated . In addition , its effects on T cell stimulatory capability was investigated by coculturing with naïve CD45RA-positive T cells . RESULTS : DB08895 decreased expression of P33681 / P42081 in a concentration-dependent manner in LPS-stimulated DCs ; however , it did not affect HLA-DR expression . DB08895 suppressed tumour necrosis factor , interleukin ( IL ) -6 and IL-1β production without affecting transforming growth factor ( TGF ) -β and P22301 production . Meanwhile , P33681 / P42081 expression in DCs was enhanced by type I interferon ( IFN ) stimulation , and the LPS-induced P33681 / P42081 expression was inhibited by an antibody to type I IFN receptor . Furthermore , tofacitinib suppressed production of type I IFN and activation of interferon regulatory factor ( Q969Q1 ) -7 , which is a transcription factor involved in P33681 / P42081 and type I IFN expression . DB08895 also decreased the T cell stimulatory capability of DCs and increased expression of indoleamine 2,3-dioxygenase ( P14902 ) -1 and Q6ZQW0 . CONCLUSIONS : DB08895 , a P23458 / P52333 inhibitor , affected the activities of human DCs . It decreased P33681 / P42081 expression and T cell stimulatory capability through suppression of type I IFN signalling . These results suggest a novel mode of action for tofacitinib and a pivotal role for JAKs in the differentiation of DCs . Loss of SHP1 enhances P52333 / P40763 signaling and decreases proteosome degradation of P52333 and P06748 - Q9UM73 in Q9UM73 + anaplastic large-cell lymphoma . Previous studies showed that most cases of Q9UM73 (+) anaplastic large-cell lymphoma ( Q9UM73 (+)ALCL ) do not express SHP1 , a tyrosine phosphatase and an important negative regulator for cellular signaling pathways such as that of JAK/ P35610 . To fully assess the biologic significance of loss of SHP1 in Q9UM73 (+)ALCL , we transfected SHP1 plasmids into 2 SHP1(-) , Q9UM73 (+)ALCL cell lines , Karpas 299 and SU-DHL-1 . After 24 hours of transfection , pJAK3 and pSTAT3 were decreased , and these changes correlated with down-regulation of P40763 downstream targets including cyclin D3 , mcl-1 , and bcl-2 . Expression of SHP1 in these 2 cell lines also resulted in marked decreases in the protein levels of P52333 and P06748 - Q9UM73 , and these effects were reversible by proteosome inhibitor MG132 . Conversely , when SHP1 expression in P60880 -M2 ( a SHP1(+) Q9UM73 (+)ALCL cell line ) was inhibited using siRNA , pSTAT3 , pJAK3 , P52333 , and P06748 - Q9UM73 were all up-regulated . Coimmunoprecipitation studies showed that SHP1 was physically associated with P52333 and P06748 - Q9UM73 . SHP1 expression in Karpas 299 and SU-DHL-1 led to significant G(1) cell cycle arrest but not apoptosis . To conclude , loss of SHP1 contributes to the pathogenesis of Q9UM73 (+)ALCL by 2 mechanisms : ( 1 ) it leaves the tyrosine phosphorylation and activation of P52333 / P40763 unchecked and ( 2 ) it decreases proteosome degradation of P52333 and P06748 - Q9UM73 . A specific inhibitor of janus kinase-3 increases survival in a transgenic mouse model of amyotrophic lateral sclerosis . Amyotrophic lateral sclerosis ( P35858 ) is a progressive , fatal neurodegenerative disorder involving the motor neurons of cortex , brain stem , and spinal cord . About 10 % of all P35858 patients are familial cases ( FALS ) , of which 20 % have mutations in the Cu , Zn-superoxide dismutase ( P00441 ) gene . The murine model for FALS , which overexpresses a FALS variant of the P00441 gene , exhibits progressive limbic paralysis followed by death . Treatment of FALS mice with WHI-P131 , a specific inhibitor of P52333 ( P52333 ) , increased survival by more than two months , suggesting that specific inhibitors of P52333 may be useful in the treatment of human P35858 . These results uniquely establish P52333 as a novel molecular target for the treatment of FALS . Preclinical to clinical translation of tofacitinib , a Janus kinase inhibitor , in rheumatoid arthritis . A critical piece in the translation of preclinical studies to clinical trials is the determination of dosing regimens that allow maximum therapeutic benefit with minimum toxicity . The preclinical pharmacokinetic ( PK ) /pharmacodynamic ( PD ) profile of tofacitinib , an oral Janus kinase ( JAK ) inhibitor , in a mouse collagen-induced arthritis ( mCIA ) model was compared with clinical PK/PD data from patients with rheumatoid arthritis ( RA ) . Preclinical evaluations included target modulation and PK/PD modeling based on continuous subcutaneous infusion or oral once- or twice-daily ( P55957 ) dosing paradigms in mice . The human PK/PD profile was obtained from pooled data from four phase 2 studies in patients with RA , and maximal effect models were used to evaluate efficacy after 12 weeks of tofacitinib treatment ( 1-15 mg P55957 ) . In mCIA , the main driver of efficacy was inhibition of cytokine receptor signaling mediated by P23458 heterodimers , but not O60674 homodimers , and continuous daily inhibition was not required to maintain efficacy . Projected efficacy could be predicted from total daily exposure irrespective of the oral dosing paradigm , with a total steady-state plasma concentration achieving 50 % of the maximal response ( Cave50 ) of ~100 nM . DB08895 potency ( ED50 ) in clinical studies was ~3.5 mg P55957 ( 90 % confidence interval : 2.3 , 5.5 ) or total Cave50 of ~40 nM , derived using Disease Activity Scores from patients with RA . The collective clinical and preclinical data indicated the importance of Cave as a driver of efficacy , rather than maximum or minimum plasma concentration ( Cmax or Cmin ) , where Cave50 values were within ~2-fold of each other . P04035 inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection . DB08895 . DB08895 ( CP-690,550 ; CP-690550 ; CP690550 ) , an orally active immunosuppressant , is being developed by Pfizer for the treatment of rheumatoid arthritis , inflammatory bowel disease , dry eyes , ankylosing spondylitis , psoriasis , psoriatic arthritis , and for the prevention of transplant rejection . DB08895 specifically inhibits Janus activated kinase 3 ( P52333 ) , which has a pivotal role in cytokine signal transduction that governs lymphocyte survival , proliferation , differentiation , and apoptosis . This review discusses the key development milestones and therapeutic trials of this drug . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . Q9NR12 -420 , a small molecular inhibitor of P01375 -α , prolongs islet allograft survival by induction of suppressor of cytokine signaling-1 : synergistic effect with cyclosporin-A . Inflammatory insults following islet transplantation ( ITx ) hinders engraftment and long-term function of the transplanted ( Tx ) islets . Using a murine model of ITx , we determined the role of Q9NR12 -420 , a novel P01375 -α inhibitor , both individually and in combination with the immunosuppressant cyclosporine A ( Q13216 ) in islet engraftment and survival . Diabetic C57BL/6 mice were Tx with 500 BALB/c islets under the kidney capsule . Four cohorts were used : Q9NR12 -420 only , Q13216 only , combination of Q9NR12 -420 and Q13216 ( Q9NR12 + Q13216 ) , and control ( n = 12 per cohort ) . Serial monitoring of blood glucose levels revealed that Q9NR12 + Q13216 ( 35 ± 5 days ) prolonged stable blood insulin levels compared to control ( 6 ± 4 days ) . Immunohistology demonstrated that coadministration ( Q9NR12 + Q13216 ) results in a significant decrease in CD8(+) T-cell infiltration ( Q9NR12 + Q13216 : 31 ± 18 vs. control : 224 ± 51 cells , p < 0.001 ) . Serum cytokine analysis revealed that Q9NR12 -420 administration resulted in an increase in the anti-inflammatory cytokine P22301 ( 2.5-fold ) , and a decrease in P01375 -α ( threefold ) with no change in P60568 . However , coadministration resulted in a marked decrease in both P60568 and P01375 -α ( threefold ) along with increase in P22301 ( threefold ) . Coadministration also demonstrated increase of antiapoptotic O15524 and Mn-SOD expression and significant reduction of donor-specific antibodies ( p < 0.005 ) . In conclusion , Q9NR12 -420 administration with Q13216 results in the upregulation of anti-inflammatory and antiapoptotic mechanisms which facilitate islet allograft engraftment and survival . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Janus kinase inhibition with tofacitinib : changing the face of inflammatory bowel disease treatment . The advent of anti-Tumor Necrosis Factor ( P01375 ) therapy has changed the way of treating inflammatory bowel disease ( Q9UKU7 ) . However , primary and secondary failure are relatively frequent with all anti- P01375 agents , which are available only as parenteral agents . DB08895 is an oral janus kinase ( JAK ) inhibitor that inhibits JAK family kinase members , in particular P23458 and P52333 , achieving a broad limitation of inflammation by interfering with several cytokine receptors . It first proved its efficacy as an immunosuppressive regimen after renal transplantation , and was recently approved by the FDA for rheumatoid arthritis . First data in Q9UKU7 are promising , especially in ulcerative colitis . Ongoing clinical trials in both UC and Crohn 's disease ( CD ) are needed to further explore its efficacy in CD and to better assess its safety profile . Microglial activation , increased P01375 and P31645 expression in the prefrontal cortex define stress-altered behaviour in mice susceptible to anhedonia . A chronic stress paradigm comprising exposure to predation , tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some , but not all , animals . Here , we sought to extend our behavioural studies to investigate how susceptibility ( sucrose preference < 65 % ) or resilience ( sucrose preference > 65 % ) to stress-induced anhedonia affects the 5HT system and the expression of inflammation-related genes . All chronically stressed animals , displayed increased level of anxiety , but susceptible mice exhibited an increased propensity to float in the forced swim test and demonstrate hyperactivity under stressful lighting conditions . These changes were not present in resilient or acutely stressed animals . Compared to resilient animals , susceptible mice showed elevated expression of tumour necrosis factor alpha ( P01375 ) and the 5-HT transporter ( P31645 ) in the pre-frontal area . Enhanced expression of 5HT(2A) and P23219 in the pre-frontal area was observed in all stressed animals . In turn , indoleamine-2,3-dioxygenase ( P14902 ) was significantly unregulated in the raphe of susceptible animals . At the cellular level , increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals . Consequently , the susceptible animals display a unique molecular profile when compared to resilient , but anxious , animals . Unexpectedly , this altered profile provides a rationale for exploring anti-inflammatory , and possibly , P01375 -targeted therapy for major depression . Inhibition of Janus kinase/signal transducer and activator of transcription ( JAK/ P35610 ) signalling pathway in rheumatoid synovial fibroblasts using small molecule compounds . Janus kinase ( JAK ) inhibitors have been developed as anti-inflammatory agents and have demonstrated clinical efficacy in rheumatoid arthritis ( RA ) . We investigated if P52333 -selective inhibition alone could disrupt cytokine signalling in rheumatoid synovial fibroblasts . In-vitro studies were performed using synovial fibroblasts isolated from patients with RA . Levels of activated JAK and signal transducer and activator of transcription ( P35610 ) proteins were detected by immunoblot analysis . Target-gene expression levels were measured by reverse transcription-polymerase chain reaction ( RT-PCR ) or real-time PCR . The JAK inhibitors CP-690,550 and INCB028050 both suppressed activation of P23458 /-2/-3 and downstream P35610 -1/-3/-5 , as well as the expression levels of target proinflammatory genes ( MCP-I , P0DJI8 /2 ) in oncostatin-M ( P13725 ) -stimulated rheumatoid synovial fibroblasts . In contrast , the P52333 -selective inhibitor , PF-956980 , suppressed P35610 -1/-5 activation but did not affect P35610 -3 activation in P13725 -stimulated rheumatoid synovial fibroblasts . In addition , PF-956980 significantly suppressed P13500 gene expression , but did not block P0DJI8 /2 gene expression in P13725 -stimulated rheumatoid synovial fibroblasts . These data suggest that P52333 -selective inhibition alone is insufficient to control P35610 -3-dependent signalling in rheumatoid synovial fibroblasts , and inhibition of JAKs , including P23458 /-2 , is needed to control the proinflammatory cascade in RA . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Tofacitinab in renal transplantation . DB08895 ( tositinib , CP-690,550 ) is a small molecule inhibitor of Janus associated kinases , primarily P52333 and O60674 , which inhibits cytokine signaling through the IL-2Rγ chain . In this article , we review the mechanism of action of tofacitinib , and pre-clinical and clinical data regarding its use in solid organ transplantation thus far . It is hoped that tofacitinib may form the basis for calcineurin-free immunosuppression , improving renal function while eliminating calcineurin inhibitor renal toxicity . Current studies suggest that tofacitinib is an effective immunosuppressive agent for renal transplantation , but it 's use in current protocols carries an increased risk of CMV , BK , and EBV viral infection , anemia and leukopenia , and post-transplant lymphoproliferative disorder . 5-Aza-2'-deoxycytidine sensitizes busulfan-resistant myeloid leukemia cells by regulating expression of genes involved in cell cycle checkpoint and apoptosis . Busulfan ( Bu ) is a DNA-alkylating drug used in myeloablative pretransplant conditioning therapy for patients with myeloid leukemia ( ML ) . A major obstacle to successful treatment is cellular Bu-resistance . To investigate the possible contribution of DNA hypermethylation to Bu-resistance , we examined the cytotoxic activity of combined DB01262 ( P22760 ) and Bu . Exposure of Bu-resistant P46977 /Bu250(6) ML cells to 0.5 microM P22760 resulted in G2-arrest and apoptosis . The observed G2-arrest was associated with hypomethylation and subsequent expression of epigenetically controlled genes including p16(INK4A) , activation of the p53 pathway , and phosphorylation of P06493 . The P22760 -mediated apoptosis was partly due to hypomethylation and up-regulation of Q6GPH4 , which resulted in down-regulation of the anti-apoptotic proteins P98170 , cIAP1 and cIAP2 . The pro-apoptotic PUMA and Q12983 proteins were up-regulated while pro-survival P40763 and c-MYC were suppressed . Combination of 0.05 microM P22760 and 5 microg/ml Bu resulted in synergistic cytotoxicity , which was associated with P09874 cleavage and activation of caspases 3 and 8 , suggesting induction of an apoptotic response . P04637 inhibition in P46977 /Bu250(6) cells using pifithrin-alpha alleviated these effects , suggesting a role for p53 therein ; this observation was supported by the relative resistance of p53-null K562 cells to [ P22760 +Bu ] combinations and by the effects of an anti-p53 shRNA on the OCI- Q13950 cell line . We conclude that the synergistic effects of [ P22760 +Bu ] are p53-dependent and involve cell cycle arrest , apoptosis induction and down-regulation of pro-survival genes . Our results suggest that , depending on tumor p53 status , incorporation of P22760 might synergistically improve the cytoreductive efficacy of Bu-based pretransplant regimen in patients with ML . Antiinflammatory and analgesic efficacy of P35354 specific inhibition : from investigational trials to clinical experience . Research strongly indicates that increased expression of the isoenzyme cyclooxygenase-2 ( P35354 ) is responsible for elevated production of prostaglandins in inflamed joint tissues and is involved in the mediation of pain . In contrast , P23219 is a constitutively produced isoenzyme that is involved in the synthesis of eicosanoids that have important homeostatic functions , for example , in the gastric mucosa and platelets . This new knowledge led to the development of drugs that are highly specific inhibitors of P35354 while not inhibiting P23219 at maximally efficacious dosages . The first P35354 specific agent approved for clinical use in the United States was celecoxib . Large multicenter trials have shown that celecoxib at dosages of 100 mg P55957 and 200 mg P55957 is as effective as naproxen 500 mg P55957 in patients with osteoarthritis of the knee or hip . Another large multicenter trial also demonstrated that celecoxib 200 mg P43251 and 400 mg P55957 is as effective as naproxen 500 mg P55957 in patients with rheumatoid arthritis ( RA ) . A comparative trial showed that celecoxib 200 mg P55957 is as effective as diclofenac SR 75 mg P55957 in patients with RA . The potential of P35354 specific inhibitors to provide antiinflammatory and analgesic efficacy equivalent to that of conventional nonsteroidal antiinflammatory drugs without the adverse gastrointestinal mucosal and platelet effects associated with nonspecific P36551 inhibitors promises to revolutionize the clinical care of arthritis patients . DB00435 and thiol redox regulation of Janus kinase activity . The activation of Janus kinases ( JAKs ) is crucial for propagation of the proliferative response initiated by many cytokines . The proliferation of various cell lines , particularly those of hematopoietic origin , is also modulated by mediators of oxidative stress such as nitric oxide and thiol redox reagents . Herein we demonstrate that nitric oxide and other thiol oxidants can inhibit the autokinase activity of rat O60674 in vitro , presumably through oxidation of crucial dithiols to disulfides within O60674 . The reduced form of O60674 is the most active form , and the oxidized O60674 form is inactive . DB00435 pretreatment of quiescent Ba/ P13726 cells also inhibits the interleukin 3-triggered in vivo activation of O60674 , a phenomenon that correlates with inhibited proliferation . Furthermore , we observed that the autokinase activity of P52333 responds in a similar fashion to thiol redox reagents in vitro and to nitric oxide donors in vivo . We suggest that the thiol redox regulation of JAKs may partially explain the generally immunosuppressive effects of nitric oxide and of other thiol oxidants . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . AZD1480 blocks growth and tumorigenesis of P07949 - activated thyroid cancer cell lines . Persistent P07949 activation is a frequent event in papillary thyroid carcinoma ( PTC ) and medullary thyroid carcinoma ( P04629 ) . In these cancers , P07949 activates the P29323 /MAPK , the PI3K/AKT/ P42345 and the JAK/ P40763 pathways . Here , we tested the efficacy of a P23458 /2- inhibitor , AZD1480 , in the in vitro and in vivo growth of thyroid cancer cell lines expressing oncogenic P07949 . Thyroid cancer cell lines harboring P07949 / Q13635 ( TPC-1 ) , P07949 M918T ( MZ-CRC1 ) and P07949 C634W ( TT ) alterations , as well as TPC-1 xenografts , were treated with JAK inhibitor , AZD1480 . This inhibitor led to growth inhibition and/or apoptosis of the thyroid cancer cell lines in vitro , as well as to tumor regression of TPC-1 xenografts , where it efficiently blocked P40763 activation in tumor and stromal cells . This inhibition was associated with decreased proliferation , decreased blood vessel density , coupled with increased necrosis . However , AZD1480 repressed the growth of P40763 - deficient TPC-1 cells in vitro and in vivo , demonstrating that its effects in this cell line were independent of P40763 in the tumor cells . In all cell lines , the JAK inhibitor reduced phospho-Y1062 P07949 levels , and P42345 effector phospho-S6 , while P23458 /2 downregulation by siRNA did not affect cell growth nor P07949 and S6 activation . In conclusion , AZD1480 effectively blocks proliferation and tumor growth of activated P07949 - thyroid cancer cell lines , likely through direct P07949 inhibition in cancer cells as well as by modulation of the microenvironment ( e.g. via JAK/phospho- P40763 inhibition in endothelial cells ) . Thus , AZD1480 should be considered as a therapeutic agent for the treatment of P07949 - activated thyroid cancers . Q14164 signals through Q14653 and NFkappaB to mediate the production of inflammatory cytokines . Q14164 and Q9UHD2 were recently identified as IKK-related kinases that are activated by toll-like receptors O15455 and O00206 . These kinases were identified as essential components of the virus-activated as well as LPS-MyD88 independent kinase complex that phosphorylates Q14653 and results in the production of cytokines involved in innate immunity . Both Q14164 and Q9UHD2 have also been implicated in the activation of the NFkappaB pathway but the precise mechanism is not clear . Although the literature to date suggests that Q14164 and Q9UHD2 play redundant roles in O15455 and O00206 signaling , recent data suggest that there may be subtle differences in the signaling pathways affected by these kinases . We have generated tetracycline-inducible stable cell lines that express a wild type or kinase-inactive mutant form of Q14164 . Our data suggest that expression of Q14164 can activate both NFkappaB and Q14653 , leading to the production of several cytokines including interferon beta . Q14164 most likely acts upstream of O14920 to activate NFkappaB in these cells since expression of the kinase-inactive version of Q14164 did not inhibit TNFalpha mediated production of inflammatory cytokines . The data suggest that Q14164 is not involved in P01375 mediated signaling but instead could likely play a role in activating O14920 downstream of Toll-like receptor signaling . We also identified P42224 , Tyk2 , and P23458 as secondary mediators of Q14164 signaling as a result of interferon beta production in these cells . [ Molecular-target therapy for advanced malignant melanoma ] . Malignant melanoma is insensitive to chemotherapy , and standard therapy for metastatic melanoma has been dacarbazine for years . Molecular abnormalities of malignant melanoma , mainly of Q96HU1 kinase signals such as P15056 mutation , have been clarified , and molecular target therapy for melanoma has been developed recently . DB08881 , an inhibitor for mutated P15056 , has shown its efficacy for the first time , with response rate of more than 50 % , and an overall improvement in survival compared with dacarbazine in a phase III study . Skin toxicities including squamous cell carcinoma , are the most severe adverse events . Another P15056 inhibitor , dabrafenib , and a MEK inhibitor , trametinib , have shown excellent efficacy in clinical studies . Melanoma also has high immunogenicity , and cytokines or cell immunotherapy have shown some efficacy . Recently , the importance of immune checkpoints which adjust T-cell activation , such as the cytotoxic T-lymphocyte-associated antigen 4 ( P16410 ) , - P33681 or the programmed cell death protein-1(PD1)-PD1 ligand( Q9NZQ7 ) , have been clarified . Targeting those immune checkpoints is expected to be effective for enhancing tumor immunity . P16410 antibody ipilimumab has been reported to improve overall survival in two phase III studies . Major adverse events were autoimmune response such as colitis , eruption , liver dysfunction and endocrineopathies . Antibodies to PD1 or Q9NZQ7 have shown a higher response rate than those of ipilimumab , and seem to accompany fewer autoimmune responses in phase I studies . These two types of targeting therapy are expected to be standard therapies for melanoma . Interaction of murine peritoneal leukocytes and mesothelial cells : in vitro model system to survey cellular events on serosal membranes during inflammation . All serosal cavities including peritoneum are lined with a simple squamous mesothelium . Primary culture of murine mesothelial cells has been established to study their cellular interactions with peritoneal leukocytes . The mesothelial character was determined by the cytokeratin and vimentin expression . The mesothelial cells expressed P05362 and P16070 molecules . The expression of P05362 , but not P16070 , was significantly enhanced by the treatment with P01375 ( 100 U/ml ) . We have also investigated possible influence of transforming growth factors , TGF-alpha ( 20 ng/ml ) and TGF-beta ( 2 ng/ml ) , and epidermal growth factor ( 20 ng/ml ) . These factors were not found to modulate P05362 or P16070 expression in vitro . During coculture experiments unstimulated mesothelial cells were almost nonadherent for both resident and elicited peritoneal mononuclear leukocytes for several hours . P01375 or P01133 pretreatment of mesothelial cells greatly enhanced their adhesive affinity to peritoneal mononuclear leukocytes , while TGF-beta pretreatment even reduced the low basal adhesion . Prolonged coculture for 3 weeks resulted in remarkable proliferation and differentiation of both resident and elicited monocytes/macrophages on the mesothelial surface . The stimulation of mesothelial cell culture with P01133 resulted in the macrophage colony-stimulating activity ( M- Q13216 ) production . M- Q13216 was mainly due to P09603 as confirmed with anti P09603 monoclonal antibody ; the residual M- Q13216 was not formed by GM- P04141 . After several passages the mesothelial cells started to produce M- Q13216 spontaneously . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . The emergence of DNA methylation as a key modulator of aberrant cell death in prostate cancer . It is now well established that cancer cells exhibit a number of genetic defects in the machinery that governs programmed cell death and that sabotage of apoptosis is one of the principal factors aiding in the evolution of the carcinogenic phenotype . A number of studies have implicated aberrant DNA methylation as a key survival mechanism in cancer , whereby promoter hypermethylation silences genes essential for many processes including apoptosis . To date , studies on the methylation profile of apoptotic genes have largely focused on cancers of the breast , colon and stomach , with only limited data available on prostate cancer . Here we discuss the major developments in the field of DNA methylation and its role in the regulation of aberrant apoptosis in prostate cancer . The most significant advances have involved the discovery of apoptotic gene targets of methylation , including Q6GPH4 , ( fragile histidine triad ( P49789 ) , cellular retinol binding protein 1 ( P09455 ) , decoy receptor 1( O14798 ) , decoy receptor 2 ( Q9UBN6 ) , target of methylation-induced silenceing 1 ( Q9ULZ3 ) , P01375 receptor superfamily , member 6 ( FAS ) , Reprimo ( Q9NS64 ) and P08151 pathogenesis-related 1 ( P48060 ) . These genes are reported to be hypermethylated in prostate cancer and some offer potential as diagnostic and prognostic markers . We also introduce the concept of an ' apoptotic methylation signature ' for prostate cancer and evaluate its potential in a diagnostic , prognostic and therapeutic setting . Signaling by retinol and its serum binding protein . DB00162 , retinol , circulates in blood bound to retinol-binding protein ( P02753 ) which , in turn , associates with transthyretin ( P02766 ) to form a retinol- P02753 - P02766 ternary complex . At some tissues , retinol-bound ( holo- ) P02753 is recognized by a membrane protein termed Q9BX79 , which transports retinol from extracellular P02753 into cells and , concomitantly , activates a O60674 / P40763 /5 signaling cascade that culminates in induction of P35610 target genes . Q9BX79 -mediated retinol transport and cell signaling are critically inter-dependent , and they both require the presence of cellular retinol-binding protein 1 ( P09455 ) , an intracellular retinol acceptor , as well as a retinol-metabolizing enzyme such as lecithin:retinol acyltransferase ( O95237 ) . Q9BX79 thus functions as a " cytokine signaling transporter " which couples vitamin A homeostasis and metabolism to cell signaling , thereby regulating gene transcription . Recent studies provided molecular level insights into the mode of action of this unique protein . Clinical utility of the oral JAK inhibitor tofacitinib in the treatment of rheumatoid arthritis . Immune/inflammatory cells act in rheumatoid arthritis ( RA ) -affected patients by synthesizing several inflammatory mediators , including cytokines that initiate intracellular signaling . Recently , small molecule inhibitors of transduction and transcription signals that influence the intracellular pathways ( such as the Janus kinase [ JAK ] family of tyrosine kinases ) have been tested for RA treatment . Four members of the JAK family are known : P23458 , O60674 , P52333 , and TyK2 . P23458 / P52333 constitutively binds to the cytoplasmic portion of the cytokine receptor - the common gamma chain - that represents a common subunit of several cytokines involved in T-cell and natural killer cell development , as well as in B-cell activation . DB08895 is an oral JAK inhibitor that is now available and effective in RA treatment , as shown in multiple Phase II and Phase III clinical trials . However , long-term safety data and comparisons with other disease-modifying antirheumatic drugs and small molecule inhibitors are necessary to better determine the role of tofacitinib in RA . DB01076 inhibits RhoC function and limits head and neck cancer metastasis . OBJECTIVE : RhoC oncogene is a well characterized marker of metastasis in a majority of invasive cancers , including HNSCC . Elevated RhoC expression has been found to be associated with distant metastasis . Statins are a class of drugs that are used to reduce cholesterol levels by inhibiting P04035 activity which in turns prevents mevalonate synthesis , which is a precursor for synthesis of cholesterol and prenylation . Interestingly , the proper function of Rho proteins depends on prenylation . Significantly , it has been reported that metastasis in human melanoma can be reduced by atorvastatin which inhibits RhoC activity by preventing its geranylgeranylation . Given that RhoC is a key oncogene involved in metastasis , we hypothesized DB01076 can reduce head and neck metastasis by inhibiting RhoC activity . METHODS : In vitro and in vivo studies were carried out to evaluate the ability of DB01076 to inhibit RhoC function and HNSCC metastasis . Cell motility , proliferation , cell invasion , and colony formation assays were performed according to the standard protocols . RESULTS : DB01076 treatment significantly reduced the active form of RhoC in vitro and diminished cell motility , invasion , proliferation and colony formation . Importantly , we observed a significant decrease in p- P27361 /2 and p- P40763 in DB01076 treated cell lines . In vivo experiments revealed inhibition of angiogenesis and lung metastases with DB01076 therapy . CONCLUSIONS : This study is the first of its kind to establish a potential role of DB01076 in head and neck cancer therapy . These findings suggest that DB01076 can be a potential low risk adjuvant therapy to minimize metastases in aggressive forms of HNSCC . EGCG enhances the therapeutic potential of gemcitabine and CP690550 by inhibiting P40763 signaling pathway in human pancreatic cancer . BACKGROUND : Signal Transducer and Activator of Transcription 3 ( P40763 ) is an oncogene , which promotes cell survival , proliferation , motility and progression in cancer cells . Targeting P40763 signaling may lead to the development of novel therapeutic approaches for human cancers . Here , we examined the effects of epigallocathechin gallate ( EGCG ) on P40763 signaling in pancreatic cancer cells , and assessed the therapeutic potential of EGCG with gemcitabine or P52333 inhibitor CP690550 ( DB08895 ) for the treatment and/or prevention of pancreatic cancer . METHODOLOGY/PRINCIPAL FINDINGS : Cell viability and apoptosis were measured by XTT assay and TUNEL staining , respectively . Gene and protein expressions were measured by qRT-PCR and Western blot analysis , respectively . The results revealed that EGCG inhibited the expression of phospho and total P52333 and P40763 , P40763 transcription and activation , and the expression of P40763 -regulated genes , resulting in the inhibition of cell motility , migration and invasion , and the induction of caspase-3 and PARP cleavage . The inhibition of P40763 enhanced the inhibitory effects of EGCG on cell motility and viability . Additionally , gemcitabine and CP690550 alone inhibited P40763 target genes and synergized with EGCG to inhibit cell viability and induce apoptosis in pancreatic cancer cells . CONCLUSIONS/SIGNIFICANCE : Overall , these results suggest that EGCG suppresses the growth , invasion and migration of pancreatic cancer cells , and induces apoptosis by interfering with the P40763 signaling pathway . Moreover , EGCG further enhanced the therapeutic potential of gemcitabine and CP690550 against pancreatic cancer . Influence of P09874 polymorphisms in patients after traumatic brain injury . Poly(ADP-ribose) polymerase-1 ( P09874 ) plays an important role in the cellular response to stress and DNA damage . However , excessive activity of P09874 exacerbates brain injury via NAD+ depletion and energy failure . The purpose of this study was to determine if tagging single nucleotide polymorphisms ( tSNPs ) covering multiple regions of the P09874 gene are related to outcome after traumatic brain injury ( TBI ) in humans . DNA from 191 adult patients with severe TBI was assayed for four tSNPs corresponding to haplotype blocks spanning the P09874 gene . Categorization as favorable or poor outcome was based on Glasgow Outcome Scale ( GOS ) score assigned at 6 months . P09874 enzyme activity was indirectly evaluated by quantifying poly-ADP-ribose ( PAR ) -modified proteins in cerebrospinal fluid ( P04141 ) using an enzyme-linked immunosorbent assay . In multiple logistic regression analysis controlling for age , initial Glasgow Coma Scale score , and gender , the AA genotype of SNP rs3219119 was an independent predictor of favorable neurologic outcome . This SNP tags a haplotype block spanning the automodification and catalytic domains of the P09874 gene . SNP rs2271347 correlated with P04141 PAR-modified protein level . This SNP , which did not correlate with outcome , tags a haplotype block spanning the promoter region of the P09874 gene . We conclude that after severe TBI in humans , a P09874 polymorphism within the automodification-catalytic domain is associated with neurological outcome , while a polymorphism within the promoter region was associated with P04141 PAR-modified protein level . These findings must be replicated in a prospective study before the relevance of P09874 polymorphisms after TBI can be established . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . P09874 deficiency blocks P05113 expression through calpain-dependent degradation of P35610 -6 in a murine asthma model . BACKGROUND : We recently showed that poly(ADP-ribose)polymerase-1 ( P09874 ) may play a role in allergen (ovalbumin)-induced airway eosinophilia , potentially through a specific effect on P05113 production . We also reported that while P05113 replenishment promotes reversal of eosinophilia in lungs of P09874 (-/-) mice , P05112 or Immunoglobulin E replenishment do not , suggesting a potentially significant regulatory relationship between P09874 and P05113 . OBJECTIVE : To explore the mechanism by which P09874 regulates P05113 production and to determine how P09874 inhibition blocks allergen-induced eosinophilia . METHODS : This study was conducted using a murine model of allergic airway inflammation and primary splenocytes . RESULTS : P09874 knockout-associated reduction in P05113 upon allergen exposure occurs at the mRNA level . Such an effect appears to take place after P05112 receptor activation as P09874 inhibition exerted no effect on P23458 / P52333 activation . Signal transducer and activator of transcription-6 ( P35610 -6 ) protein was severely downregulated in spleens of P09874 (-/-) mice without any effect on mRNA levels , suggesting an effect on protein integrity rather than gene transcription . Interestingly , the degradation of P35610 -6 in P09874 (-/-) mice required allergen stimulation . Additionally , P09874 enzymatic activity appears to be required for P35610 -6 integrity . The downregulation of P35610 -6 coincided with mRNA and protein reduction of GATA-binding protein-3 and occupancy of its binding site on the P05113 gene promoter . P05112 was sufficient to induce P35610 -6 downregulation in both P09874 (-/-) mice and isolated splenocytes . Such degradation may be mediated by calpain , but not by proteasomes . CONCLUSION : These results demonstrate a novel function of P09874 in regulating P05113 expression during allergen-induced inflammation and explain the underlying mechanism by which P09874 inhibition results in P05113 reduction . Novel treatments with small molecules in psoriatic arthritis . Current treatment options for patients with active psoriatic arthritis ( PsA ) include synthetic disease-modifying antirheumatic drugs and biologic agents . Propelled by increased understanding of immunopathogenesis of PsA , new therapeutic agents targeting different biologic pathways have been evaluated . This article discusses novel small-molecule , orally available treatments that are currently in clinical development for the treatment of psoriasis and PsA . This includes the phosphodiesterase 4 inhibitor apremilast and Janus kinase ( JAK ) inhibitors . Apremilast has demonstrated significant improvements in patients with moderate to severe psoriasis and PsA in phase II and III clinical trials and has recently been approved for the treatment of PsA . DB08895 , an oral inhibitor of P52333 , P23458 , and , to a lesser degree , O60674 , approved for the treatment of rheumatoid arthritis in several countries , has demonstrated positive results in psoriasis in phase II studies . Studies in PsA are ongoing . With these new developments , treatment options will continue to improve in the future . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . JAK inhibitors : treatment efficacy and safety profile in patients with psoriasis . Janus kinase ( JAK ) pathways are key mediators in the immunopathogenesis of psoriasis . Psoriasis treatment has evolved with the advent of targeted therapies , which inhibit specific components of the psoriasis proinflammatory cascade . JAK inhibitors have been studied in early phase trials for psoriasis patients , and the data are promising for these agents as potential treatment options . DB08895 , an oral or topically administered P23458 and P52333 inhibitor , and ruxolitinib , a topical P23458 and O60674 inhibitor , have been most extensively studied in psoriasis , and both improved clinical symptoms of psoriasis . Additional P23458 or P52333 inhibitors are being studied in clinical trials . In phase III trials for rheumatoid arthritis , tofacitinib was efficacious in patients with inadequate responses to tumor necrosis factor inhibitors , methotrexate monotherapy , or disease-modifying antirheumatic drugs . The results of phase III trials are pending for these therapies in psoriasis , and these agents may represent important alternatives for patients with inadequate responses to currently available agents . Further investigations with long-term clinical trials are necessary to verify their utility in psoriasis treatment and assess their safety in this patient population . Kinase inhibitors : a new class of antirheumatic drugs . The outlook for patients with rheumatoid arthritis has improved significantly over the last three decades with the use of disease-modifying antirheumatic drugs . However , despite the use of methotrexate , cytokine inhibitors , and molecules targeting T and B cells , a percentage of patients do not respond or lose their response over time . The autoimmune process in rheumatoid arthritis depends on activation of immune cells , which utilize intracellular kinases to respond to external stimuli such as cytokines , immune complexes , and antigens . In the past decade , small molecules targeting several kinases , such as p38 MAPK , Syk , and JAK have been developed . Several p38 MAPK inhibitors proved ineffective in treating rheumatoid arthritis . The Syk inhibitor , fostamatinib , proved superior to placebo in Phase II trials and is currently under Phase III investigation . DB08895 , a P23458 /3 inhibitor , was shown to be efficacious in two Phase III trials , while VX-509 , a P52333 inhibitor , showed promising results in a Phase II trial . Fostamatinib and tofacitinib were associated with increased rates of infection , elevation of liver enzymes , and neutropenia . Moreover , fostamatinib caused elevations of blood pressure and diarrhea , while tofacitinib was associated with an increase in creatinine and elevation of lipid levels . DB08895 in kidney transplantation . INTRODUCTION : This review will discuss the mechanism of action and important kidney transplant clinical trial data for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . AREAS COVERED : Successful kidney transplantation requires adequate immunosuppression . Current maintenance immunosuppressive protocols which rely on calcineurin inhibitors have long-term nephrotoxicity and negative impact on cardiometabolic risk factors . JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared to control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . EXPERT OPINION : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . P23219 -derived thromboxane A2 plays an essential role in early B-cell development via regulation of JAK/ P42229 signaling in mouse . Cyclooxygenases ( COXs ) and their prostanoid products play important roles in a diverse range of physiological processes , including in the immune system . Here , we provide evidence that P23219 is an essential regulator in early stages of B-cell development . P23219 -deficient mice displayed systematic reduction in total B cells , which was attributed to the arrest of early B-cell development from pro-B to pre-B stage . We further demonstrated that this defect was mediated through downregulation of the Janus kinase/signal transducer and activator of transcription 5 ( JAK/ P42229 ) signaling and its target genes , including Pax5 , in P23219 (-/-) mice . Mechanistic studies revealed that P23219 -derived thromboxane A2 ( TxA2 ) could regulate P52333 / P42229 signaling through the cyclic adenosine monophosphate-protein kinase A pathway , via binding with its receptor thromboxane A2 receptor ( TP ) . Administration of the TP agonist could rescue the defective B-cell development and JAK/ P42229 signaling activity in P23219 -deficient mice . Moreover , administration of low-dose aspirin caused a significant reduction in total B cells in peripheral blood of healthy human volunteers , coincidentally with reduced TxA2 production and downregulation of JAK/ P42229 signaling . Taken together , our results demonstrate that P23219 -derived TxA2 plays a critical role in the stage transition of early B-cell development through regulation of JAK/ P42229 signaling and indicate a potential immune-suppressive effect of low-dose aspirin in humans . Efficacy and safety of tofacitinib for treatment of rheumatoid arthritis . DB08895 is the first in a new class of nonbiologic disease-modifying antirheumatic drugs ( DMARDs ) , a targeted , synthetic DMARD , approved for the treatment of rheumatoid arthritis ( RA ) as monotherapy or in combination with methotrexate or other non-biologic DMARD . DB08895 , an orally administered Janus kinase ( JAK ) inhibitor , decreases T-cell activation , pro-inflammatory cytokine production , and cytokine signaling by inhibiting binding of type I cytokine receptors family and γ-chain cytokines to paired P23458 / P52333 receptors . The net effect of tofacitinb 's mechanism of action is decreased synovial inflammation and structural joint damage in RA patients . To date , six phase 3 trials have been conducted to evaluate the safety and efficacy of tofacitinib under the oral rheumatoid arthritis triaLs ( ORAL ) series . This review describes the pharmacology of the novel agent , tofacitinib , and details the safety and efficacy data of the ORAL trials .

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DB02709 inhibits PDGF receptor mitogenic signaling in mesangial cells : role of P18031 . Mesangioproliferative glomerulonephritis is associated with overactive PDGF receptor signal transduction . We show that the phytoalexin resveratrol dose dependently inhibits PDGF-induced DNA synthesis in mesangial cells with an IC(50) of 10 microM without inducing apoptosis . Remarkably , the increased Q96EB6 deacetylase activity induced by resveratrol was not necessary for this inhibitory effect . DB02709 significantly blocked PDGF-stimulated c-Src and Akt kinase activation , resulting in reduced cyclin D1 expression and attenuated P06400 phosphorylation and cyclin-dependent kinase-2 ( P24941 ) activity . Furthermore , resveratrol inhibited P09619 phosphorylation at the PI 3 kinase and Grb-2 binding sites tyrosine-751 and tyrosine-716 , respectively . This deficiency in P09619 phosphorylation resulted in significant inhibition of PI 3 kinase and Erk1/2 MAPK activity . Interestingly , resveratrol increased the activity of protein tyrosine phosphatase P18031 , which dephosphorylates PDGF-stimulated phosphorylation at tyrosine-751 and tyrosine-716 on P09619 with concomitant reduction in Akt and Erk1/2 kinase activity . P18031 significantly inhibited PDGF-induced DNA synthesis without inducing apoptosis . These results for the first time provide evidence that the stilbene resveratrol targets P18031 to inhibit P09619 mitogenic signaling . P01344 Producing Hepatocellular Carcinoma Treated with DB00398 : Metabolic Complications and a Foresight to Molecular Targeting Therapy to the IGF Signal . Hypoglycemia is a rare paraneoplastic manifestation of patients with neoplasms . Hypoglycemia can be induced by several causes , including an aberrant increase of hypoglycemic agents and adrenal insufficiency . DB00398 is the first agent to demonstrate a survival benefit in the treatment of advanced hepatocellular carcinoma ( HCC ) . This small molecule inhibits serine/threonine kinase RAF in tumor cells and tyrosine kinases VEGFR/ P09619 in tumor vasculature and decreases tumor growth and angiogenesis . In this paper , we report a case of HCC who was treated with sorafenib and showed severe hypoglycemia . This hypoglycemia might be induced by two causes , both adrenal insufficiency as an adverse effect of sorafenib and activation of the insulin-like growth factor ( IGF ) signal by excessive secretion of incompletely processed precursors of P01344 . Although the IGF signal is suggested to be involved in aberrant growth of HCC in some cases , there is no other report showing the influence of sorafenib on HCC with active IGF signal . Unfortunately , the effect of sorafenib was limited in the present case . However , emerging drugs that directly inhibit the IGF signal can be expected to be highly effective in the treatment of HCC with hypoglycemia . Integrin alpha5-induced P00533 activation by prothrombin triggers hepatocyte apoptosis via the JNK signaling pathway . We have previously shown that prothrombin , a blood coagulation factor , can cause an inhibition of DNA synthesis in normal rat hepatocytes . To explore the mechanisms of this prothrombin action , we examined its effects on the activation of fibronectin receptor integrin alpha5 , since fibronectin was found to be degraded by prothrombin actions in primary hepatocyte cultures . We found that prothrombin treatment of rat hepatocytes without addition of any growth factor induced tyrosine phosphorylation of integrin alpha5 and interaction of integrin alpha5 with epidermal growth factor receptor ( P00533 ) , leading to P00533 tyrosine phosphorylation at tyrosine residues DB00135 -845 and DB00135 -1173 . P00533 tyrosine phosphorylation triggered phosphorylation of its down-stream target Shc and the activation of the c-Jun N-terminal kinase ( JNK ) pathway . P00734 also induced hepatocyte apoptosis , a change in cell shape and activation of caspase 3 pathway . The JNK pathway is most likely involved in prothrombin-induced hepatocyte apoptosis , because pre-treatment of hepatocytes with JNK kinase inhibitor II ( SP600125 ) antagonized these prothrombin actions . The data suggest that integrin-related P00533 activation by prothrombin can induce cell growth inhibition and apoptosis via an P00533 -JNK signaling pathway . [ Eruptive nevi associated with sorafenib treatment ] . BACKGROUND : DB00398 is a new multikinase inhibitor recently approved for renal cell carcinoma and hepatocarcinoma . Among other targets , it blocks the kinase function of the RAF gene products including V600E mutant P15056 , which is frequently found in both melanoma and naevi . Cutaneous side effects are frequent with sorafenib , but no naevus modification has been reported until now . PATIENTS AND METHODS : Five cases of eruptive naevi in patients treated with sorafenib are reported . The mean duration of sorafenib treatment was 9.2 months when naevi eruption was noticed . The patients presented with about 100 to more than 200 small , homogenous , dark-brown naevi located mainly on the trunk and upper limbs . DISCUSSION : Eruptive melanocytic naevi have been reported in association with blistering diseases , and more generally in a setting of immunosuppression . We hypothesize that naevi appearance could be linked to an anti-senescence effect of sorafenib via its action on the Q96HU1 kinase pathway . Further prospective studies are needed to explore the relationship between sorafenib and the biology of naevi . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . P01308 -like growth factor-II mediates the steroidogenic and growth promoting actions of follicle stimulating hormone on human ovarian pre-antral follicles cultured in vitro . DB00094 is important for ovarian antral follicle growth and steroidogenesis , processes in the human that are believed to be mediated by P01344 . The objective of this study was to determine if human ovarian pre-antral follicles are also DB00094 - and P01344 -responsive , since the clinical questions and mechanisms underlying the effects on the pre-antral follicle pool of exogenously administered gonadotropins for fertility therapy and elevated endogenous gonadotropins in the perimenopause remain unanswered . Class 2 preantral follicles were isolated from human premenopausal ovaries ( n=6 ) and cultured in vitro with androstenedione and either no additives or with DB00094 or P01344 . DB00094 ( 100 ng/mL ) stimulated estradiol ( E2 ) production by 3.58 +/- 0.4 fold over 48 hr , compared to controls without DB00094 . This effect was completely inhibited in the presence of the P01344 antagonist , IGF binding protein-4 ( P22692 ) . P01344 also stimulated E2 production by preantral follicles with doses as low as 1 ng/mL and within 24 hr of treatment . Maximal response of 3- to 9-fold above control was achieved with 100 ng/mL of P01344 between 96-120 hr of culture . P22692 completely inhibited E2 production to basal levels . DB00094 stimulated P01344 mRNA in pre-antral follicles about 4-fold , determined by RT-PCR . DB00094 also stimulated follicle growth , determined by light microscopy , 50-68 % over 48 hr , compared to controls ( P < 0.001 ) , a process that was inhibited in the presence of P22692 . Cumulatively , these data support P01344 as a mediator of DB00094 action on human preantral follicles . DB00398 Induced Hand Foot Skin Rash in P36888 ITD Mutated Acute Myeloid Leukemia-A Case Report and Review of Literature . DB00398 is a novel small molecule multiple kinase inhibitor which has been used for metastatic renal cancer , hepatocellular cancer . DB00398 induced skin rash has been discussed as a side effect in trials in both , P36888 wild type and mutated acute myeloid leukemia ( AML ) , as monotherapy or as combination with other chemotherapeutic agents . We describe a patient with P17948 3 ITD mutated AML , who was started on adjunctive DB00398 therapy . Skin reactions manifested as NCI Grade III palmoplantar erythrodysesthesia ( PPE ) , requiring drug discontinuation . Several pathogenic mechanisms have been implicated in DB00398 induced skin reactions , but none has been conclusively proven . While treatment options are varied for early stage skin reactions , drug discontinuation remains the only possible therapy presently for severe grade skin reaction . Targeting PIM kinases impairs survival of hematopoietic cells transformed by kinase inhibitor-sensitive and kinase inhibitor-resistant forms of P36888 and P11274 / P00519 . Previous studies have shown that activation of the signal transducer and activator of transcription 5 ( P42229 ) plays an essential role in leukemogenesis mediated through constitutive activated protein tyrosine kinases ( PTK ) . Because PIM-1 is a P42229 target gene , we analyzed the role of the family of PIM serine/threonine kinases ( PIM-1 to PIM-3 ) in PTK-mediated transformation of hematopoietic cells . Ba/ P13726 cells transformed to growth factor independence by various oncogenic PTKs ( P41212 / O60674 , P41212 / Q16288 , P41212 / P00519 , P11274 / P00519 , P36888 -ITD , and H4/PDGFbetaR ) show abundant expression of PIM-1 and PIM-2 . Suppression of PIM-1 activity had a negligible effect on transformation . In contrast , expression of kinase-dead PIM-2 mutant ( PIM-2KD ) led to a rapid decline of survival in Ba/ P13726 cells transformed by P36888 -ITD but not by other oncogenic PTKs tested . Coexpression of PIM-1KD and PIM-2KD abrogated growth factor-independent growth of Ba/ P13726 transformed by several PTKs , including P11274 / P00519 . Targeted down-regulation of PIM-2 by RNA interference ( RNAi ) selectively abrogated survival of Ba/ P13726 cells transformed by various P36888 ( P36888 ) -activating mutants [ internal tandem duplication ( ITD ) and kinase domain ] and attenuated growth of human cell lines containing P36888 mutations . Interestingly , cells transformed by P36888 and P11274 / P00519 mutations that confer resistance to small-molecule tyrosine kinase inhibitors were still sensitive to knockdown of PIM-2 , or PIM-1 and PIM-2 by RNAi . Our observations indicate that combined inactivation of PIM-1 and PIM-2 interferes with oncogenic PTKs and suggest that PIMs are alternative therapeutic targets in PTK-mediated leukemia . Targeting the PIM kinase family could provide a new avenue to overcome resistance against small-molecule tyrosine kinase inhibitors . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Cutaneous drug eruptions induced by sorafenib : a case series . DB00398 , an epidermal growth factor receptor inhibitor , is a novel treatment used for malignancies resistant to traditional chemotherapy . Epidermal growth factor receptors ( P00533 ) are a family of 4 transmembrane tyrosine kinase receptors that , via signal transduction pathways , mediate cell growth , differentiation , and survival . DB00398 is a targeted drug specifically engineered to inhibit Raf serine/threonine kinases , which are part of the reticular activating system ( DB01367 ) oncogene pathway . In addition , in vitro studies have shown sorafenib to be a potent multikinase inhibitor , targeting receptor tyrosine kinases associated with tumor angiogenesis ( P35968 , P35916 , and P09619 ) and progression . Initially , approved for use in advanced renal cell carcinoma , sorafenib is being studied for the treatment of other solid tumors at our institution . During the clinical trial , 4 patients were referred to the dermatology clinic for evaluation and treatment of diffuse erythematous eruptions all occurring 8 to 10 days after initiating sorafenib at a dose of 400 mg twice daily . These eruptions occurred in demographically similar patients and displayed similar clinical characteristics and histopathological findings . Clinically , 3 of 4 patients had facial erythema , 3 of 4 had generalized macular erythema , 3 of 4 had widespread follicular-based papular eruption , and 4 of 4 had palmoplantar erythrodysesthesia . Half of the patients had cutaneous eruptions without systemic effects , while the other half had hypersensitivity reactions requiring withdrawal from clinical trial . This is the first case series illustrating drug eruptions induced by sorafenib . DB00398 inhibits many kinase mutations associated with drug-resistant gastrointestinal stromal tumors . DB00398 has substantial clinical activity as third- or fourth-line treatment of imatinib- and sunitinib-resistant gastrointestinal stromal tumors ( GIST ) . Because sorafenib targets both angiogenesis-related kinases ( VEGFR ) and the pathogenetic kinases found in GIST ( P10721 or P16234 ) , the molecular basis for sorafenib efficacy in this setting remains unknown . We sought to determine the spectrum of activity of sorafenib against different mutant kinases associated with drug-sensitive and drug-resistant GIST . We compared the activity of imatinib and sorafenib against transiently expressed mutant forms of P10721 and P16234 , including various secondary mutations that have been identified in imatinib-resistant or sunitinib-resistant GISTs . We also examined these drugs against four GIST cell lines , three of which are imatinib resistant . In our in vitro studies , we determined that sorafenib inhibited imatinib-resistant mutations in exons encoding the DB00171 /drug-binding pocket and in exons encoding the activation loop , with the exception of substitutions at P10721 codon D816 and P16234 codon 842 . Notably our data indicate that sorafenib is more effective than imatinib or sunitinib for inhibiting the kinase activity of drug-resistant P10721 mutants ( as assessed by biochemical IC(50) ) . We hypothesize that a major determinant of the efficacy of sorafenib for treatment of advanced GIST is the activity of this agent against P10721 or P16234 -mutant kinases . These results have implications for the further development of treatments for drug-resistant GIST . Off-label use of cetuximab plus sorafenib and panitumumab plus regorafenib to personalize therapy for a patient with V600E P15056 -mutant metastatic colon cancer . DB00398 , the first agent developed to target P15056 mutant melanoma , is a multi-kinase inhibitor that was approved by the FDA for therapy of kidney and subsequently liver cancer , and is currently in clinical trials for thyroid , lung and brain cancer . Colorectal cancer with V600E P15056 mutation has shown relative resistance to standard chemotherapy regimens , as well as lack of efficacy to vemurafenib in clinical trials . New treatments are needed for P15056 -mutant colorectal cancer . We report a case of a patient with P15056 -mutant metastatic colon cancer whose disease had progressed on FOLFOX plus bevacizumab and subsequent FOLFIRI plus cetuximab . Based on preclinical data published in Nature in 2012 suggesting that successful therapeutic targeting of P15056 in colorectal cancer may require concomitant targeting of the P00533 , we offered this patient without other attractive options the combination of sorafenib plus cetuximab , in off-label use with informed consent . DB00398 and cetuximab therapy led to a mixed radiographic response with some areas showing dramatic improvement and other areas showing stable disease over a 7-month period which is a notably long period of progression-free survival for V600E P15056 mutated colon cancer . The cetuximab plus sorafenib therapy was very well-tolerated by the patient who remained on it long enough until another therapy option , regorafenib , was approved in September 2012 . The patient was offered single agent regorafenib at the time of progression . At the time of progression on single agent regorafenib , panitumumab was combined with regorafenib and this was also well-tolerated and appeared to slow disease progression . Further study of these approaches in the clinic as personalized treatment of P15056 -mutant advanced colorectal cancer is warranted . A preclinical evaluation of a novel multikinase inhibitor , SKLB-329 , as a therapeutic agent against hepatocellular carcinoma . Hepatocellular carcinoma ( HCC ) is a serious life-threatening malignant disease of liver . Molecular targeted therapies are considered a promising strategy for the treatment of HCC . DB00398 is the first , and so far the only targeted drug approved by the US Food and Drug Administration ( FDA ) for clinical therapy of HCC . Despite being effective in some HCC patients , some demerits of sorafenib in the treatment of HCC , such as modest survival benefits , and drug resistance , have also been reported , which highlights the unmet medical need among patients with HCC . Here , we report a novel multikinase inhibitor discovered by us , SKLB-329 , which potently inhibits angiogenesis-related kinases including P17948 /2/3 , and P21802 , and the Src kinase . SKLB-329 significantly inhibited endothelial cell growth , migration , invasion and tube formation . It showed potent anti-angiogenic activity in a transgenic zebrafish model . Moreover , SKLB-329 could efficiently restrain the proliferation of HCC cells through down-regulation of Src-mediated Q05397 and Stat3 activity . In vivo , oral administration of SKLB-329 considerably suppressed the tumor growth in HCC xenograft models ( HepG2 and SMMC7721 ) in a dose-dependent manner . In all of the in vitro and in vivo assays of this investigation , sorafenib was used as a positive control , and in most assays SKLB-329 exhibited a higher potency compared with the positive control . In addition , SKLB-329 also bears favorable pharmacokinetic properties . Collectively , the results of preclinical studies presented here demonstrate that SKLB-329 is a promising drug candidate for HCC treatment . [ Hand-foot syndrome and sorafenib ] . DB00398 ( Nexavar ) is a targeted therapy acting as VEGFR and P09619 tyrosine-kinase inhibitor that has been approved in France in the treatment of metastatic renal cell carcinoma and hepatocarcinoma . Hand-foot syndrome is one of the more frequent toxicity related to sorafenib . This paper up lights the main points concerning this toxicity in the view of specialists working together in the care of these patients : a pharmacologist , a dermatologist and a medical oncologist . The clinic and symptoms of hand-foot syndrome as the biological interpretation , the symptomatic treatment and the impact on the specific treatment of sorafenib are developed . Ectopic DB01285 syndrome caused by bronchial carcinoid tumor indistinguishable from Cushing 's disease . A 75-year-old woman was admitted to our hospital because of a poor glycemic control . She was found to have Cushingoid feature and dynamic endocrine tests showed elevated plasma DB01285 and cortisol levels , lack of their circadian rhythm , non-suppressibility to high-dose dexamethasone , responsiveness to P06850 , but not to DB00035 , and suppression to octreotide . Pituitary Q9BWK5 showed an equivocal small lesion . CT scan of the chest showed two nodular lesions in the right lung ( S5 , S7 ) , while a mild uptake was noted only in S5 lesion by DB09150 -PET , but positive uptake was only in S7 lesion by somatostatin receptor scintigraphy ( SRS ) . Inferior petrosal sinus sampling revealed a gradient of plasma DB01285 after P06850 stimulation , consistent with the diagnosis of Cushing ' s disease . She underwent middle and inferior lobectomy of the right lung . The resected tumor in S7 was consistent with the diagnosis of a bronchial carcinoid tumor with positive DB01285 immunoreactivity , while that of S5 was cryptococcal granuloma . RT-PCR revealed abundant expressions of P01189 and SSTR ( -1 , -2 , -5 ) , but not of P34998 and P47901 . Postoperatively , abnormal endocrine data were normalized along with improvement of hypertension and diabetes . This was a diagnostic challenging case with ectopic DB01285 syndrome indistinguishable from Cushing ' s disease by various endocrine and imaging tests , among which SRS successfully localized the tumor responsible for ectopic DB01285 secretion . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . Functionalisation of PLLA nanofiber scaffolds using a possible cooperative effect between collagen type I and P12643 : impact on growth and osteogenic differentiation of human DB05914 . Mesenchymal stem cell differentiation of osteoblasts is triggered by a series of signaling processes including integrin and bone morphogenetic protein ( BMP ) , which therefore act in a cooperative manner . The aim of this study was to analyze whether these processes can be remodeled in an artificial poly-(L)-lactide acid ( PLLA ) based nanofiber scaffold . Matrices composed of PLLA-collagen type I or P12643 incorporated PLLA-collagen type I were seeded with human DB05914 ( hMSC ) and cultivated over a period of 22 days , either under growth or osteoinductive conditions . During the course of culture , gene expression of alkaline phosphatase ( ALP ) , osteocalcin ( OC ) and collagen I ( COL-I ) as well as Q99717 and focal adhesion kinase ( Q05397 ) , two signal transduction molecules involved in P12643 or integrin signaling were analyzed . Furthermore , calcium and collagen I deposition , as well as cell densities and proliferation , were determined using fluorescence microscopy . The incorporation of P12643 into PLLA-collagen type I nanofibers resulted in a decrease in diameter as well as pore sizes of the scaffold . Mesenchymal stem cells showed better adherence and a reduced proliferation on BMP-containing scaffolds . This was accompanied by an increase in gene expression of ALP , OC and COL-I . Furthermore the presence of P12643 resulted in an upregulation of Q05397 , while collagen had an impact on the gene expression of Q99717 . Therefore these different strategies can be combined in order to enhance the osteoblast differentiation of hMSC on PLLA based nanofiber scaffold . By doing this , different signal transduction pathways seem to be up regulated . Serotonin transporter interacts with the PDGFβ receptor in DB00102 -induced signaling and mitogenesis in pulmonary artery smooth muscle cells . The serotonin transporter ( P31645 ) and the platelet-derived growth factor receptor ( P09619 ) have been implicated in both clinical and experimental pulmonary hypertension ( PH ) and the facilitation of pulmonary artery smooth muscle cell ( PASMC ) growth . To gain a better understanding of the possible relationship of these two cell surface molecules we have explored interactions between P31645 and P09619 . We have previously demonstrated that P31645 transactivates PDGFRβ in serotonin-stimulated PASMC proliferation . We now provide evidence for a role for P31645 in DB00102 signaling and PASMC proliferation by using pharmacological inhibitors , genetic ablation , and construct overexpression of P31645 . The results show that four tested P31645 blockers dose dependently inhibit PDGF-stimulated human and bovine PASMC proliferation with comparable efficacy to that of P09619 inhibitors , whereas P28222 or 5- Q13049 receptor inhibitors had no effect . Combinations of the P31645 and P09619 inhibitors led to synergistic/additive inhibition . Similarly , PDGF-induced PASMC proliferation was attenuated by small interfering RNA downregulation of P31645 . Inhibition of P31645 in PASMCs attenuated PDGF-induced phosphorylation of PDGFRβ , Akt , and p38 but not Erk . Overexpression of P31645 in HEK293 cells led to enhanced Akt phosphorylation by PDGF , which was blunted by a P31645 PDZ motif mutant , indicating the mechanistic need for the PDZ motif of P31645 in PDGF signaling . Furthermore , coimmunoprecipitation experiments showed that P31645 and PDGFRβ become physically associated upon PDGF stimulation . In total , the data show for the first time an important interactive relationship between P31645 and the PDGFRβ in the production of PASMC proliferation triggered by PDGF that may be important in PH . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Human endothelial progenitor cells isolated from P48444 patients are dysfunctional . Cardiovascular disease is the leading cause of morbidity and mortality in patients with moderate-to-severe chronic obstructive pulmonary disease ( P48444 ) . More than 44 % of these patients present with generalized atherosclerosis at autopsy . It is accepted that endothelial progenitor cells ( EPCs ) participate in the repair of dysfunctional endothelium and thus protects against atherosclerosis . However , whether P48444 affects the repairing capacity of EPCs is unknown . Therefore , the objective of this study was to determine whether and how EPCs are involved in the vascular repair process in patients with P48444 . In our study , EPCs from 25 P48444 and 16 control patients were isolated by Ficoll density-gradient centrifugation and identified using fluorescence activated cell sorting . Transwell Migratory Assay was performed to determine the number of EPC colony-forming units and the adherent capacity late-EPCs to human umbilical vein endothelial cells . Following arterial damage in NOD/SCID mice , the number of EPCs incorporated at the injured vascular site was determined using a fluorescence microscope . We found that the number of EPC clusters and cell migration , as well as the expression of P61073 , was significantly decreased in patients with P48444 . Additionally , the number of late-EPCs adherent to HUVEC tubules was significantly reduced , and fewer P35968 (+)-staining cells were incorporated into the injured site in P48444 patients . Our study demonstrates that EPC capacity of repair was affected in P48444 patients , which may contribute to altered vascular endothelium in this patient population . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Q02548 / P41212 acts as a transcriptional repressor causing down-modulation of P15391 , enhances migration to P48061 , and confers survival advantage in pre-BI cells . Q02548 is a transcription factor essential for B-cell development . Recently , it has been found as a frequent target of aberrancies in childhood acute lymphoblastic leukemia ( ALL ; 30 % of B cell ALL cases ) , showing monoallelic loss , point mutations , or chromosomal translocations . The role of these aberrancies is still poorly understood . We previously cloned the Q02548 / P41212 fusion gene in a patient affected by B-cell precursor ALL with a t(9;12) translocation . This is the first report investigating the molecular and functional roles of Q02548 / P41212 protein in vitro from murine wild-type pre-BI cells . We showed that Q02548 / P41212 protein acts as an aberrant transcription factor with repressor function , recruiting mSin3A , down-regulating B220 , P15391 , Q8WV28 , MB-1 , P36888 , and mu heavy chain expression , thus suggesting a block on B-cell differentiation . In a Q02548 -deficient context , the presence of Q02548 / P41212 did not replace Q02548 functions . Q02548 / P41212 protein enhances cell migration towards P48061 , with the overexpression of P61073 . Moreover , the presence of the fusion gene overcomes interleukin-7 withdrawal and interferes with transforming growth factor-beta1 pathway , inducing resistance and conferring cells an advantage in proliferation and survival . Thus , in vitro , the Q02548 / P41212 protein has a dominant effect on wild-type Q02548 , interferes with the process of B-cell differentiation and migration , and induces resistance to apoptosis . Taken together , these phenomena likely represent key events in the process of B-cell transformation . DB00398 as monotherapy or in association with cytarabine and clofarabine for the treatment of relapsed/refractory P36888 ITD-positive advanced acute myeloid leukemia . DB00398 ( BAY 43-9006 ) in hepatocellular carcinoma patients : from discovery to clinical development . Angiogenesis and signaling through the DB01367 /RAF/mitogen-activated protein/extracellular signal-regulated kinase ( P29323 ) kinase ( MEK ) / P29323 cascade have been reported to play important roles in the development of hepatocellular carcinoma ( HCC ) . DB00398 ( Nexavar ) , a novel bi-aryl urea BAY 43-9006 , is an orally administered multikinase inhibitor with activity against DB01367 /RAF kinases multikinase inhibitor with activity against RAF kinases and several receptor tyrosine kinases , including vascular endothelial growth factor receptor ( VEGFR ) , platelet-derived growth factor receptor ( P09619 ) , P36888 , Ret , and c-Kit . It is involved in angiogenic pathway and cell proliferation . DB00398 has demonstrated potent anti-tumor activity in in vitro studies , preclinical xenograft models of different tumor types and human clinical trials . This review summarizes the history of sorafenib from its discovery by the medicinal chemistry approach through to clinical development and ongoing trials on the combination between sorafenib and trans-arterial chemoembolization ( P78536 ) in HCC patients . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Importance of MEK in neutrophil microbicidal responsiveness . Exposure of neutrophils to inflammatory stimuli such as the chemoattractant FMLP leads to activation of responses including cell motility , the oxidative burst , and secretion of proteolytic enzymes . A signaling cascade involving sequential activation of P04049 , mitogen-activated protein kinase ( MEK ) , and extracellular signal regulated kinase ( P29323 ) is also rapidly activated after agonist exposure . The temporal relationship between these events suggests that the kinases may be involved in triggering the effector functions , but direct evidence of a causal relationship is lacking . To assess the role of the MEK/ P29323 pathway in the activation of neutrophil responses , we studied the effects of PD098059 , a potent and selective inhibitor of MEK . Preincubation of human neutrophils with 50 microM PD098059 almost completely ( > 90 % ) inhibited the FMLP-induced activation of MEK-1 and MEK-2 , the isoforms expressed by neutrophils . This dose of PD098059 virtually abrogated chemoattractant-induced tyrosine phosphorylation and activation of P27361 and P28482 , implying that MEKs are the predominant upstream activators of these mitogen-activated protein kinases . Pretreatment of neutrophils with the MEK antagonist inhibited the oxidative burst substantially and phagocytosis only moderately . In addition , PD098059 antagonized the delay of apoptosis induced by exposure to granulocyte-macrophage P04141 . However , the effects of PD098059 were selective , as it failed to inhibit other responses , including chemoattractant-induced exocytosis of primary and secondary granules , polymerization of F-actin , chemotaxis , or activation of phospholipase A2 . We conclude that MEK and P29323 contribute to the activation of the oxidative burst and phagocytosis , and participate in cytokine regulation of apoptosis . DB00398 : recent update on activity as a single agent and in combination with interferon-alpha2 in patients with advanced-stage renal cell carcinoma . Metastatic renal cell carcinoma does not respond favorably to conventional treatment strategies and is not very responsive to cytokine therapy . Therefore , novel targeted treatment approaches have been explored for patients with renal cancer who have chemotherapy-refractory disease . DB00398 ( BAY 43-9006 ) is a small-molecule inhibitor that has been shown to target members of multiple classes of tyrosine kinases that are known to be involved in tumor cell proliferation and tumor angiogenesis . These kinases include vascular endothelial growth factor receptor ( VEGFR ) -1 , P35968 , P35916 , platelet-derived growth factor receptor , Flt-3 , c-kit , and Raf kinases . Based on the significant improvement in progression-free survival , sorafenib received Food and Drug Administration approval in December 2005 for the treatment of renal cell carcinoma . In combination studies , sorafenib with other antitumor agents has demonstrated significant clinical activity in patients with renal cell carcinoma . As discussed in this mini-review , the clinical potency of sorafenib as a single agent or in combination with other antitumor agents is being evaluated in several ongoing clinical trials in patients with renal carcinoma . Putative role of HIF transcriptional activity in melanocytes and melanoma biology . Hypoxia-inducible factor-1α ( HIF-1α ) is a highly oxygen sensitive bHLH protein that is part of the heterodimeric Q9BYW2 transcription factor . Under hypoxic stress , Q9BYW2 activity is induced to control expression of multiple downstream target genes , including vascular endothelial growth factor ( P15692 ) . The normal epidermis exists in a constant mild hypoxic microenvironment and constitutively expresses HIF-1α and HIF-2α . Expression of HIF-1α and/or HIF-2α has been suggested to correlate with the increased malignant potential of melanocytes , therefore , failures of melanoma therapies may be partially linked to high HIF activity . Notably , melanomas that have the V600E P15056 mutation exhibit increased HIF-1α expression . We have utilized a bioinformatics approach to identify putative hypoxia response elements ( HREs ) in a set of genes known to participate in the process of melanogenesis ( includingTRPM1 , Q9UMX9 , P01112 , C- P10721 , P40967 and P06850 ) . While some of the mechanistic links between these genes and the HIF pathway have been previously explored , others await further investigation . Although agents targeting HIF activity have been proposed as novel treatment modalities for melanoma , there are currently no clinical trials in progress to test their efficacy in melanoma . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . Molecular mechanisms of gastrin-dependent gene regulation . The peptide hormone gastrin is the key regulator of gastric acid secretion . P01350 exerts its effects as acid secretagogue through functional activation of gastric enterochromaffin-like ( ECL ) cells , which control acid secretion through biosynthesis and release of histamine . In ECL cells , concerted activation of histidine decarboxylase ( HDC ) , vesicular monoamine transporter 2 ( Q05940 ) , and chromogranin A ( P10645 ) genes by gastrin is a prerequisite for proper acid control . To elucidate the molecular pathways underlying gastrin-dependent control of ECL cell genes , we recently analyzed the signaling cascades , regulatory promoter elements , and transcription factors mediating the transcriptional effects of gastrin . Our studies identified the Raf > Q02750 > P29323 1/-2 kinase module as the common signaling pathway mediating gastrin-dependent ECL cell gene transcription . In contrast to this uniform signaling cascade , pronounced heterogeneity was detected between cis- and trans-activating regulatory factors conferring gastrin responsiveness . The molecular diversity of transcription factors and regulatory enhancer elements transmitting gastrin-triggered gene transcription offers the molecular basis for synergistic , but differential , regulation of HDC , Q05940 , and P10645 genes during a secretory challenge of ECL cells by gastrin and possibly other acid secretagogues . DB00398 suppresses JNK-dependent apoptosis through inhibition of Q9NYL2 . DB00398 is U.S . Food and Drug Adminstration-approved for the treatment of renal cell carcinoma and hepatocellular carcinoma and has been combined with numerous other targeted therapies and chemotherapies in the treatment of many cancers . Unfortunately , as with other RAF inhibitors , patients treated with sorafenib have a 5 % to 10 % rate of developing cutaneous squamous cell carcinoma ( cSCC ) /keratoacanthomas . Paradoxical activation of extracellular signal-regulated kinase ( P29323 ) in P15056 wild-type cells has been implicated in RAF inhibitor-induced cSCC . Here , we report that sorafenib suppresses UV-induced apoptosis specifically by inhibiting c-jun-NH(2)-kinase ( JNK ) activation through the off-target inhibition of leucine zipper and sterile alpha motif-containing kinase ( Q9NYL2 ) . Our results implicate suppression of JNK signaling , independent of the P29323 pathway , as an additional mechanism of adverse effects of sorafenib . This has broad implications for combination therapies using sorafenib with other modalities that induce apoptosis . DB00398 : complexities of Raf-dependent and Raf-independent signaling are now unveiled . Hepatocellular carcinoma ( HCC ) is the most common primary cancer worldwide . The only current drug available for clinical treatment of HCC is sorafenib , which inhibits multiple signaling kinases including Raf family members , platelet-derived growth factor receptor , vascular endothelial growth factor receptors 1 and 2 , c-Kit , and P36888 . Many studies have revealed that the mechanism underlying the antitumor effect of sorafenib is complex . Because sorafenib inhibits C-Raf more potently than B-Raf , the therapeutic efficacy of sorafenib is strongly influenced by the relative expression and activity of B-Raf and C-Raf and the complex interactions between these factors . Moreover , Rafindependent signaling mechanisms have recently emerged as important pathways of sorafenib-induced cell death . Basic research studies have suggested that using sorafenib as part of a combination therapy may improve its effect , although this has yet to be confirmed by clinical evidence . Further studies of the functional mechanism of sorafenib are required to advance the development of targeted therapy for HCC . To aid future work on sorafenib , we here review the current literature pertaining to sorafenib signaling and its clinical efficacy in both monotherapy and combination therapy . A phase I trial of the oral , multikinase inhibitor sorafenib in combination with carboplatin and paclitaxel . PURPOSE : This study evaluated the safety , maximum tolerated dose , pharmacokinetics , and antitumor activity of sorafenib , a multikinase inhibitor , combined with paclitaxel and carboplatin in patients with solid tumors . PATIENTS AND METHODS : Thirty-nine patients with advanced cancer ( 24 with melanoma ) received oral sorafenib 100 , 200 , or 400 mg twice daily on days 2 to 19 of a 21-day cycle . All patients received carboplatin corresponding to AUC6 and 225 mg/m(2) paclitaxel on day 1 . Pharmacokinetic analyses were done for sorafenib on days 2 and 19 of cycle 1 and for paclitaxel on day 1 of cycles 1 and 2 . Pretreatment tumor samples from 17 melanoma patients were analyzed for P15056 mutations . RESULTS : DB00398 was well tolerated at the doses evaluated . The most frequent severe adverse events were hematologic toxicities ( grade 3 or 4 in 33 patients , 85 % ) . Twenty-seven ( 69 % ) patients had sorafenib-related adverse events , the most frequent of which were dermatologic events ( 26 patients , 67 % ) . Exposure to paclitaxel was not altered by intervening treatment with sorafenib . Treatment with sorafenib , paclitaxel , and carboplatin resulted in one complete response and nine partial responses , all among patients with melanoma . There was no correlation between P15056 mutational status and treatment responses in patients with melanoma . CONCLUSIONS : The recommended phase II doses are oral 400 mg twice daily sorafenib , carboplatin at an AUC6 dose , and 225 mg/m(2) paclitaxel . The tumor responses observed with this combined regimen in patients with melanoma warrant further investigation . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Reinvestigation of carrier transport properties in liquid crystalline 2-phenylbenzothiazole derivatives . We have reinvestigated the charge carrier transport properties in a liquid crystal of 2-(4'-heptyloxyphenyl)-6-dodecylthiobenzothiazole ( 7O- P10721 - P28222 ) , for which the electronic conduction was first established in rodlike liquid crystals and for which the highest hole mobility in the smectic A ( SmA ) phase ever achieved was reported . We found that 7O- P10721 - P28222 exhibited three crystal phases , one of which appeared in a limited temperature range of 10 degrees just below the phase transition temperature from the SmA phase . In this crystal phase , nondispersive transient photohole currents were observed in time-of-flight experiments , and its hole mobility was determined to be 8 x 10(-3) cm(2)/Vs , slightly higher than that reported previously in the SmA phase . For the SmA phase , however , the hole mobility was 1 x 10(-4) cm(2)/Vs . Furthermore , we established the electron transport in the SmA phase of purified 7O- P10721 - P28222 , whose mobility was the same as the hole mobility in that phase . In order to confirm generality of the new findings in 7O- P10721 - P28222 , we investigated the carrier transport properties of its derivative having a short hydrocarbon chain , 2-(4'-heptyloxyphenyl)-6-butylthiobenzothiazole ( 7O- P10721 -S4 ) , and obtained comparable results . The present results correct a mistake in the previous report and give an idea of what a typical mobility in the SmA phase is . On the basis of these results , we discuss what determines the charge carrier mobility in smectic mesophases . Induction of high mobility group box 1 release from serotonin-stimulated human umbilical vein endothelial cells . High mobility group box 1 ( P09429 ) is a non-histone nuclear protein which is released from the nucleus of activated macrophages into the extracellular space in response to stimuli such as endotoxin or necrosis . The P09429 functions as a potent proinflammatory cytokine in the extracellular spaces . Recently , P09429 has been implicated in the progression of atherosclerosis . However , the association between P09429 and the development of atherosclerosis is poorly understood . Therefore , we examined whether serotonin ( 5-HT ) , a key factor involved in the development of atherosclerosis , induced P09429 release in human umbilical vein endothelial cells ( HUVECs ) . We found that 5-HT induced the release of P09429 but not of P27361 /2 and JNK from HUVECs via the 5-HT receptor ( P28222 ) /p38 mitogen-activated protein kinase ( MAPK ) signaling pathway . The p38MAPK inhibitor SB203580 and the P28222 antagonist GR55526 markedly inhibited P09429 release from 5-HT-stimulated HUVECs . The vascular endothelial growth factor ( P15692 ) derived from activated macrophages in atherosclerotic lesions also plays an important role in the progression of atherosclerosis . We found that P09429 induced P15692 production in macrophage-like RAW264.7 cells . P09429 induced the activation of p38MAPK , P27361 /2 and Akt . The P19957 -kinase inhibitor LY294002 significantly inhibited P15692 production in P09429 -stimulated macrophages , while other kinase inhibitors did not . These results suggest that P09429 release may contribute as a risk factor in the development and progression of atherosclerosis . DB00398 enhances pemetrexed cytotoxicity through an autophagy-dependent mechanism in cancer cells . DB00642 ( ALIMTA ) is a folate anti-metabolite that has been approved for the treatment of non-small cell lung cancer , and has been shown to stimulate autophagy . In the present study , we sought to further understand the role of autophagy in the response to pemetrexed and to test if combination therapy could enhance the level of toxicity through altered autophagy in tumor cells . The multikinase inhibitor sorafenib ( NEXAVAR ) , used in the treatment of renal and hepatocellular carcinoma , suppresses tumor angiogenesis and promotes autophagy in tumor cells . We found that sorafenib interacted in a greater than additive fashion with pemetrexed to increase autophagy and to kill a diverse array of tumor cell types . Tumor cell types that displayed high levels of cell killing after combination treatment showed elevated levels of AKT , P08133 S6K and/or phosphorylated P42345 , in addition to class III RTKs such as PDGFRb and P17948 , known in vivo targets of sorafenib . In xenograft and in syngeneic animal models of mammary carcinoma and glioblastoma , the combination of sorafenib and pemetrexed suppressed tumor growth without deleterious effects on normal tissues or animal body mass . Taken together , the data suggest that premexetred and sorafenib act synergistically to enhance tumor killing via the promotion of a toxic form of autophagy that leads to activation of the intrinsic apoptosis pathway , and predict that combination treatment represents a future therapeutic option in the treatment of solid tumors . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Biological evaluation of a novel sorafenib analogue , t-CUPM . DB00398 ( Nexavar® ) is currently the only FDA-approved small molecule targeted therapy for advanced hepatocellular carcinoma . The use of structural analogues and derivatives of sorafenib has enabled the elucidation of critical targets and mechanism(s) of cell death for human cancer lines . We previously performed a structure-activity relationship study on a series of sorafenib analogues designed to investigate the inhibition overlap between the major targets of sorafenib P04049 kinase and P35968 , and an enzyme shown to be a potent off-target of sorafenib , soluble epoxide hydrolase . In the current work , we present the biological data on our lead sorafenib analogue , t-CUPM , demonstrating that this analogue retains cytotoxicity similar to sorafenib in various human cancer cell lines and strongly inhibits growth in the NCI-60 cell line panel . Co-treatment with the pan-caspase inhibitor , Z-VAD-FMK , failed to rescue the cell viability responses of both sorafenib and t-CUPM , and immunofluorescence microscopy shows similar mitochondrial depolarization and apoptosis-inducing factor release for both compounds . These data suggest that both compounds induce a similar mechanism of caspase-independent apoptosis in hepatoma cells . In addition , t-CUPM displays anti-proliferative effects comparable to sorafenib as seen by a halt in G0/ P55008 in cell cycle progression . The structural difference between sorafenib and t-CUPM significantly reduces inhibitory spectrum of kinases by this analogue , and pharmacokinetic characterization demonstrates a 20-fold better oral bioavailability of t-CUPM than sorafenib in mice . Thus , t-CUPM may have the potential to reduce the adverse events observed from the multikinase inhibitory properties and the large dosing regimens of sorafenib . DB00398 and thyroid cancer . DB00398 ( Nexavar ) is a multikinase inhibitor , which has demonstrated both anti-proliferative and anti-angiogenic properties in vitro and in vivo , inhibiting the activity of targets present in the tumor cell [ c-RAF ( proto-oncogene serine/threonine-protein kinase ) , P15056 , (V600E) P15056 , c- P10721 , and P07333 -like tyrosine kinase 3 ] and in tumor vessels ( c-RAF , vascular endothelial growth factor receptor-2 , vascular endothelial growth factor receptor-3 , and platelet-derived growth factor receptor β ) . For several years , sorafenib has been approved for the treatment of hepatocellular carcinoma and advanced renal cell carcinoma . After previous studies showing that sorafenib was able to inhibit oncogenic P07949 mutants , (V600E) P15056 , and angiogenesis and growth of orthotopic anaplastic thyroid cancer xenografts in nude mice , some clinical trials demonstrated the effectiveness of sorafenib in advanced thyroid cancer . Currently , the evaluation of the clinical safety and efficacy of sorafenib for the treatment of advanced thyroid cancer is ongoing . This article reviews the anti-neoplastic effect of sorafenib in thyroid cancer . Several completed ( or ongoing ) studies have evaluated the long-term efficacy and tolerability of sorafenib in patients with papillary and medullary aggressive thyroid cancer . The results suggest that sorafenib is a promising therapeutic option in patients with advanced thyroid cancer that is not responsive to traditional therapeutic strategies . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . BMP receptor-integrin interaction mediates responses of vascular endothelial Q15797 /5 and proliferation to disturbed flow . BACKGROUND : Vascular endothelial cells ( ECs ) are constantly exposed to blood flow-induced shear stress . Our previous study demonstrated that disturbed flow with low and oscillatory shear stress ( OSS ) induces bone morphogenetic protein receptor ( BMPR ) -specific Q15797 /5 activation in ECs , but the underlying mechanisms and the in vivo functional role of Q15797 /5 remain unclear . OBJECTIVES : Here we elucidated the molecular mechanisms by which OSS activates EC Q15797 /5 and its in vivo functional role . METHODS : Lentiviral Q99717 -specific short hairpin RNA ( Lenti-shSmad5 ) was constructed and intra-arterially injected into the lumen of stenosed abdominal aorta in bromodeoxyuridine-infused rats . Co-immunoprecipitation and in situ proximity ligation assays were performed on ECs exposed to OSS ( 0.5 ± 4 dynes/cm(2) ) in a parallel-plate flow chamber to investigate BMPR-integrin interactions and their regulatory role in OSS-activation of EC Q15797 /5 . RESULTS : Intra-arterial administration of Lenti-shSmad5 inhibited bromodeoxyuridine uptake of ECs at post-stenotic sites , where disturbed flow with OSS occurs . OSS induced sustained BMPRIB-αv β3 integrin association in ECs , which was mediated by the intracytoplasmic kinase domain of BMPRII and subsequently activated the Shc/focal adhesion kinase ( Q05397 ) /extracellular signal-regulated kinase ( P29323 ) cascade , leading to Q15797 /5 activation . This OSS-activation of Q15797 /5 induced its association with and activation of runt-related transcription factor-2 ( Runx2 ) , leading to activations of mammalian target of rapamycin ( P42345 ) and p70S6 kinase ( p70S6K ) , a pathway critical for EC proliferation in response to OSS . CONCLUSIONS : Oscillatory shear stress induces synergistic interactions between specific BMPRs and integrin to activate Q15797 /5 through the Shc/ Q05397 / P29323 pathway , which leads to the activation of the Runx2/ P42345 /p70S6K pathway to promote EC proliferation .

MH_train_1028

MH_train_1028

MH_train_1028

interacts_with DB09036?

multiple_choice

FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Antitumor efficacy of the anti-interleukin-6 ( P05231 ) antibody siltuximab in mouse xenograft models of lung cancer . INTRODUCTION : P05231 ( P05231 ) can activate downstream signaling pathways in lung cancer cells , such as the P40763 pathway , and is reported to be produced by tumor cells with activating P00533 mutations . We examined P05231 / P40763 in lung cancer tumor tissues and the effects of siltuximab , a neutralizing antibody to human P05231 , in mouse models of lung cancer . METHODS : P05231 and P40763 activation levels were compared with tumor histology and presence of P01116 mutations in snap-frozen , non-small-cell lung cancer tumors . The effects of siltuximab alone or in combination with erlotinib were examined in mouse xenograft models constructed using three cell line xenograft models and one primary explant mouse model . We examined the influence of cancer-associated fibroblasts ( CAFs ) on tumor growth and siltuximab effects . RESULTS : P05231 levels were higher in tumors of squamous cell versus adenocarcinoma histology and were not associated with presence of P01116 mutations . Tyrosine phosphorylation status of P40763 did not correlate with tumor P05231 levels . DB00133 phosphorylation of P40763 was correlated with P01116 mutation status . Both tumor and stromal cells contributed to total P05231 within tumors . DB09036 had minimal effect as a single agent in xenografts with tumor cells alone ; however , in models coadministered with CAFs , siltuximab had more potent effects on tumor inhibition . We observed no effects of combined erlotinib and siltuximab . CONCLUSIONS : P05231 is elevated in subsets of human NSCLCs , especially with squamous cell histology . Tumors supported by stromal production of P05231 seem to be the most vulnerable to tumor growth inhibition by siltuximab . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma . Signalling through the interleukin ( IL ) -6 pathway induces proliferation and drug resistance of multiple myeloma cells . We therefore sought to determine whether the P05231 -neutralizing monoclonal antibody siltuximab , formerly CNTO 328 , could enhance the activity of melphalan , and to examine some of the mechanisms underlying this interaction . DB09036 increased the cytotoxicity of melphalan in KAS-6/1 , Q16352 -6 , ANBL-6 , and RPMI 8226 human myeloma cell lines ( HMCLs ) in an additive-to-synergistic manner , and sensitized resistant RPMI 8226.LR5 cells to melphalan . These anti-proliferative effects were accompanied by enhanced activation of drug-specific apoptosis in HMCLs grown in suspension , and in HMCLs co-cultured with a human-derived stromal cell line . DB09036 with melphalan enhanced activation of caspase-8 , caspase-9 , and the downstream effector caspase-3 compared with either of the single agents . This increased induction of cell death occurred in association with enhanced Bak activation . Neutralization of P05231 also suppressed signalling through the phosphoinositide 3-kinase/Akt pathway , as evidenced by decreased phosphorylation of Akt , P08133 S6 kinase and Q13541 . Importantly , the siltuximab/melphalan regimen demonstrated enhanced anti-proliferative effects against primary plasma cells derived from patients with myeloma , monoclonal gammopathy of undetermined significance , and amyloidosis . These studies provide a rationale for translation of siltuximab into the clinic in combination with melphalan-based therapies . Effects of siltuximab on the P05231 -induced signaling pathway in ovarian cancer . PURPOSE : To explore potential therapeutic strategies for interrupting the interleukin-6 ( P05231 ) signaling pathway , we measured P05231 expression in ovarian cancer tissues , and evaluated the effects of a monoclonal anti- P05231 antibody ; siltuximab ( CNTO 328 ) , on levels of P05231 -induced Stat3 phosphorylation , Stat3 nuclear translocation , and Stat3 downstream antiapoptotic genes . We then looked for enhancing paclitaxel sensitivity in multidrug-resistant ovarian cancer cell lines . EXPERIMENTAL DESIGN : Expressions of P05231 in ovarian cancer patient specimens were assessed by immunohistochemistry . Effects of siltuximab on P05231 -induced activation of Stat3 in an ovarian cancer cell line were determined by Western blot and real-time analysis of Stat3 nucleocytoplasmic translocation . Influence of combination of siltuximab and paclitaxel on tumor growth was evaluated in a xenograft mouse mode in vivo . RESULTS : Metastatic and drug-resistant recurrent tumors have significantly higher P05231 expression when compared with the matched primary tumors . DB09036 specifically suppressed P05231 -induced Stat3 phosphorylation and Stat3 nuclear translocation . Treatment with siltuximab significantly decreased the levels of Stat3 downstream proteins such as Q8WXI8 -1 , Bcl-X(L) , and survivin . Treatment with siltuximab reduced expression of multiple P05231 -induced genes in these cell lines . Furthermore , siltuximab increased the cytotoxic effects of paclitaxel in a paclitaxel resistant ovarian cancer cell line in vitro , but combination therapy with siltuximab did not have a significant effect on paclitaxel resistant tumor growth in vivo . CONCLUSIONS : These results show that siltuximab effectively block the P05231 signaling pathways and P05231 -induced gene expression . Blockage of P05231 signaling may provide benefits for the treatment of ovarian cancer . A phase 2 , randomized , double-blind , placebo-controlled study of siltuximab ( anti- P05231 mAb ) and bortezomib versus bortezomib alone in patients with relapsed or refractory multiple myeloma . We compared the safety and efficacy of siltuximab ( S ) , an anti-interleukin-6 chimeric monoclonal antibody , plus bortezomib ( B ) with placebo ( plc ) + B in patients with relapsed/refractory multiple myeloma in a randomized phase 2 study . DB09036 was given by 6 mg/kg IV every 2 weeks . On progression , B was discontinued and high-dose dexamethasone could be added to S/plc . Response and progression-free survival ( PFS ) were analyzed pre-dexamethasone by European Group for Blood and Marrow Transplantation ( EBMT ) criteria . For the 281 randomized patients , median PFS for S + B and plc + B was 8.0 and 7.6 months ( HR 0.869 , P = 0.345 ) , overall response rate was 55 versus 47 % ( P = 0.213 ) , complete response rate was 11 versus 7 % , and median overall survival ( OS ) was 30.8 versus 36.8 months ( HR 1.353 , P = 0.103 ) . Sustained suppression of P02741 , a marker reflective of inhibition of interleukin-6 activity , was seen with S + B . DB09036 did not affect B pharmacokinetics . DB09036 /placebo discontinuation ( 75 versus 66 % ) , grade ≥3 neutropenia ( 49 versus 29 % ) , thrombocytopenia ( 48 versus 34 % ) , and all-grade infections ( 62 versus 49 % ) occurred more frequently with S + B . The addition of siltuximab to bortezomib did not appear to improve PFS or OS despite a numerical increase in response rate in patients with relapsed or refractory multiple myeloma . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . Dopamine agonists upregulate P05231 and P10145 production in human keratinocytes . AIM : Catecholamines regulate functions of the nervous , neuroendocrine and immune systems . Dopamine may modulate the activity of keratinocytes , which play a role in secreting cytokines and chemokines . The aim of this study was to evaluate the effect of dopaminergic agonists on the production of P05231 and P10145 by a non-tumoral human keratinocyte cell line ( HaCaT ) . METHODS : Cells were stimulated with dopamine and the P14416 agonist cabergoline . Levels of P05231 and P10145 in culture supernatants were then determined . Cell proliferation was also assessed . Assays were carried out in the presence or absence of the dopaminergic and β-adrenergic receptor antagonists ( sulpiride and propranolol , respectively ) and ascorbic acid . RESULTS : Dopamine stimulated the production of P05231 and P10145 in a concentration-dependent manner . The effects observed on the secretion of P05231 were more potent than those corresponding to P10145 and were reduced by ascorbic acid . The dopamine-induced P05231 secretion was partially reduced by sulpiride and abrogated by propranolol . The latter drug was able to block the effect of dopamine on the secretion of P10145 . The cabergoline-induced P05231 release was reduced by sulpiride . Cell viability was not affected by any of the drugs . CONCLUSIONS : Dopaminergic agonists can stimulate keratinocytes to produce P05231 and P10145 which are related to inflammatory cutaneous processes . These effects are mediated by dopaminergic and β-adrenergic receptors and by receptor-independent oxidative mechanisms . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . Th 1 cytokine production by peripheral blood mononuclear cells in X-linked adrenoleukodystrophy . Cerebral adrenoleukodystrophy ( P33897 ) and adrenomyeloneuropathy ( Q9BXJ7 ) are the two most frequent clinical phenotypes of the same genetic defect leading to the accumulation of very long chain fatty acids ( VLCFA ) . Previous studies have suggested that inflammatory cytokines may play a role in the cerebral demyelination and in phenotype expression of the disease . We analyzed cytokine production by stimulated peripheral blood mononuclear cells ( PBMC ) from 17 patients ( four asymptomatic subjects , eight Q9BXJ7 and five P33897 ) . Our results show that lipopolysaccarides ( LPS ) stimulated PBMC from both symptomatic and asymptomatic patients have an increased production of IL-12 and TNFalpha compared to controls , while after phitoemoagglutinin ( PHA ) stimulation we observed a decreased production of P05231 and P22301 . These data indicate that , following an immunological stimulus , PBMC from patients have an increased production of cytokines typical of a Th1 cell response which is able to promote the inflammatory process . This characteristic profile of cytokine production could be related to the biochemical defect and could have a role in central nervous system ( CNS ) pathogenesis . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . P23458 activates P40763 activity in non-small-cell lung cancer cells and P05231 neutralizing antibodies can suppress P23458 - P40763 signaling . Members of the signal transducer and activator of transcription ( P35610 ) family of transcription factors are potential targets for the treatment and prevention of cancers including non-small-cell lung cancer . P35610 proteins can be phosphorylated and activated by diverse upstream kinases including cytokine receptors and tyrosine kinases . We examined P35610 protein activation in lung cancer cell lines including those with activating mutations in the P00533 and examined upstream kinases responsible for P40763 phosphorylation and activation using small molecules , antibodies , and RNA interference . We found more pronounced P40763 activation in cells with activating P00533 mutations , yet inhibition of P00533 activity had no effect on P40763 activation . Inhibition of P23458 with small molecules or RNA interference resulted in loss of P40763 tyrosine phosphorylation and inhibition of cell growth . An interleukin-6 neutralizing antibody , siltuximab ( CNTO 328 ) could inhibit P40763 tyrosine phosphorylation in a cell-dependent manner . DB09036 could completely inhibit P40763 tyrosine phosphorylation in H1650 cells , and this resulted in inhibition of lung cancer cell growth in vivo . Combined P00533 inhibition with erlotinib and siltuximab resulted in dual inhibition of both tyrosine and serine P40763 phosphorylation , more pronounced inhibition of P40763 transcriptional activity , and translated into combined effects on lung cancer growth in a mouse model . Our results suggest that P23458 is responsible for P40763 activation in lung cancer cells and that indirect attacks on P23458 - P40763 using an P05231 neutralizing antibody with or without P00533 inhibition can inhibit lung cancer growth in lung cancer subsets . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified . DB09036 : first global approval . The anti-interleukin-6 ( P05231 ) chimeric monoclonal antibody siltuximab is the first drug to be approved for the treatment of multicentric Castleman 's disease ( O95822 ) in the US and European union ( EU ) , having gained approval under the FDA priority review program in the US and from an accelerated assessment and recommendation by the Committee for Medicinal Products for Human Use ( CHMP ) in the EU . Development of the drug is continuing in smoldering multiple myeloma . This article summarizes the milestones in the development of siltuximab leading to this first approval for O95822 . Modulation of Q8IVT5 activity in Caenorhabditis elegans by Zn ions , P25116 kinase and PP2A phosphatase . Vulval differentiation in Caenorhabditis elegans is controlled by a conserved signal transduction pathway mediated by Ras and a kinase cascade that includes Raf , Mek and MAPK . Activation of this cascade is positively regulated by a number of proteins such as Q8IVT5 ( kinase suppressor of Ras ) , Q09428 -8/ Q5T124 -2 , Q09428 -6/PP2A-B and P05231 -1 . We describe the functional characterization of sur-7 and several genes that regulate signaling downstream of ras . We identified sur-7 by isolating a mutation that suppresses an activated ras allele , and showed that Q09428 -7 is a divergent member of the cation diffusion facilitator family of heavy metal ion transporters that is probably localized to the endoplosmic recticulum membrane and regulates cellular Zn(2+) concentrations . Genetic double mutant analyses suggest that the Q09428 -7-mediated effect is not a general toxic response . Instead , Zn(2+) ions target a specific step of the pathway , probably regulation of the scaffolding protein Q8IVT5 . Biochemical analysis in mammalian cells indicates that high Zn(2+) concentration causes a dramatic increase of Q8IVT5 phosphorylation . Genetic analysis also indicates that PP2A phosphatase and P25116 kinase act downstream of Raf to positively and negatively regulate Q8IVT5 activity , respectively . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . DB09036 for multicentric Castleman disease . Dysregulated secretion of P05231 plays a pivotal role in the pathogenesis of Castleman disease ( CD ) , a rare lymphoproliferative disorder . In contrast to unicentric CD for which surgery is considered the treatment of choice , there is no standard therapeutic approach for multicentric CD ( O95822 ) . DB09036 ( trade name : Sylvant , formerly known as CNTO 328 ) is a chimeric monoclonal antibody with high binding affinity for human P05231 . In a recent randomized placebo-controlled Phase II trial , subjects with HIV-negative , HHV8-negative O95822 who received siltuximab demonstrated a significantly higher rate of durable tumor and symptomatic response with a tolerable safety profile , leading to its approval for the treatment of HIV-negative HHV8-negative O95822 by the US FDA and the European Commission in April and May 2014 , respectively . This article will cover the current treatment options of O95822 , the drug profile of siltuximab and future directions in the management of O95822 . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Targeting interleukin-6 in inflammatory autoimmune diseases and cancers . P05231 ( P05231 ) is a pleiotropic cytokine with significant functions in the regulation of the immune system . As a potent pro-inflammatory cytokine , P05231 plays a pivotal role in host defense against pathogens and acute stress . However , increased or deregulated expression of P05231 significantly contributes to the pathogenesis of various human diseases . Numerous preclinical and clinical studies have revealed the pathological roles of the P05231 pathway in inflammation , autoimmunity , and cancer . Based on the rich body of studies on biological activities of P05231 and its pathological roles , therapeutic strategies targeting the P05231 pathway are in development for cancers , inflammatory and autoimmune diseases . Several anti- P05231 / P05231 receptor monoclonal antibodies developed for targeted therapy have demonstrated promising results in both preclinical studies and clinical trials . DB06273 , an anti- P05231 receptor antibody , is effective in the treatment of various autoimmune and inflammatory conditions notably rheumatoid arthritis . It is the only P05231 pathway targeting agent approved by the regulatory agencies for clinical use . DB09036 , an anti- P05231 antibody , has been shown to have potential benefits treating various human cancers either as a single agent or in combination with other chemotherapy drugs . Several other anti- P05231 -based therapies are also under clinical development for various diseases . P05231 antagonism has been shown to be a potential therapy for these disorders refractory to conventional drugs . New strategies , such as combination of P05231 blockade with inhibition of other signaling pathways , may further improve P05231 -targeted immunotherapy of human diseases . Inhibitors of DB00171 -binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages . DB00171 -binding cassette ( DB01048 ) transporters are a large family of proteins whose role is to translocate various substances across biological membranes . They include the Tangier disease protein ABC1 , sulfonylurea receptors ( Q09428 ) , multidrug resistance protein ( MDR ) , and cystic fibrosis transmembrane regulator ( P13569 ) . In the current study , we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide ( LPS ) and/or interferon ( IFN ) -gamma-induced interleukin ( IL ) -12 p40 and tumor necrosis factor ( P01375 ) -alpha production , nitric oxide formation , as well as major histocompatibility complex II up-regulation in macrophages . The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production . However , glibenclamide failed to affect the production of P01375 . The selective ABC1 inhibitors 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production . On the other hand , both the MDR inhibitor verapamil and P13569 blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40 . Furthermore , selective inhibitors and activators of SURs were without effect . In agreement with the pharmacological data , macrophages expressed mRNA for ABC1 , but not SURs or P13569 . Intracellular levels of IL-12 p40 were decreased by glibenclamide , suggesting that glibenclamide does not affect IL-12 p40 secretion . The effect of glibenclamide did not involve an interference with the activation of the p38 and Q8NFH3 /44 mitogen-activated protein kinases or c-Jun kinase . DB01016 also suppressed P01579 -induced up-regulation of major histocompatibility complex II . Taken together , our results indicate that ABC proteins regulate LPS and/or P01579 -induced macrophage activation . The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor . The aryl hydrocarbon receptor ( P35869 ) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways , such as xenobiotic metabolism and Th17 cell differentiation . Identification of key physiologically relevant ligands that regulate P35869 function remains to be accomplished . Screening of indole metabolites has identified indoxyl 3-sulfate ( I3S ) as a potent endogenous ligand that selectively activates the human P35869 at nanomolar concentrations in primary human hepatocytes , regulating transcription of multiple genes , including P04798 , P05177 , Q16678 , P22309 , P19224 , P05231 , and P0DJI8 . Furthermore , I3S exhibits an approximately 500-fold greater potency in terms of transcriptional activation of the human P35869 relative to the mouse P35869 in cell lines . Structure-function studies reveal that the sulfate group is an important determinant for efficient P35869 activation . This is the first phase II enzymatic product identified that can significantly activate the P35869 , and ligand competition binding assays indicate that I3S is a direct P35869 ligand . I3S failed to activate either CAR or O75469 . The physiological importance of I3S lies in the fact that it is a key uremic toxin that accumulates to high micromolar concentrations in kidney dialysis patients , but its mechanism of action is unknown . I3S represents the first identified relatively high potency endogenous P35869 ligand that plays a key role in human disease progression . These studies provide evidence that the production of I3S can lead to P35869 activation and altered drug metabolism . Our results also suggest that prolonged activation of the P35869 by I3S may contribute to toxicity observed in kidney dialysis patients and thus represent a possible therapeutic target . P05231 sensitizes prostate cancer to the antiproliferative effect of IFNα2 through Q00978 . Development and progression of prostate cancer ( PCa ) are associated with chronic inflammation . The cytokine interleukin 6 ( P05231 ) can influence progression , differentiation , survival , and angiogenesis of PCa . To identify novel pathways that are triggered by P05231 , we performed a gene expression profiling of two PCa cell lines , LNCaP and MDA PCa 2b , treated with 5 ng/ml P05231 . Interferon ( IFN ) regulatory factor 9 ( Q00978 ) was identified as one of the most prevalent P05231 -regulated genes in both cell lines . Q00978 is a mediator of type I IFN signaling and acts together with P42224 and 2 to activate transcription of IFN-responsive genes . The P05231 regulation of Q00978 was confirmed at mRNA and protein levels by quantitative real-time PCR and western blot respectively in both cell lines and could be blocked by the anti- P05231 antibody DB09036 . Three PCa cell lines , PC3 , Du-145 , and LNCaP- P05231 + , with an autocrine P05231 loop displayed high expression of Q00978 . A tissue microarray with 36 PCa tissues showed that Q00978 protein expression is moderately elevated in malignant areas and positively correlates with the tissue expression of P05231 . Downregulation and overexpression of Q00978 provided evidence for an IFN-independent role of Q00978 in cellular proliferation of different PCa cell lines . Furthermore , expression of Q00978 was essential to mediate the antiproliferative effects of IFNα2 . We concluded that P05231 is an inducer of Q00978 expression in PCa and a sensitizer for the antiproliferative effects of IFNα2 . P28335 receptor involvement in female rat lordosis behavior . Adult , hormone-primed , ovariectomized rats ( P05231 -344 ) with bilateral implants within the ventromedial nucleus of the hypothalamus ( VMN ) , were injected with 0.5 microgram estradiol benzoate followed 48 h later with 500 microgram progesterone . This priming produced rats with 2 different levels of sexual receptivity . Rats with a lordosis to mount ratio ( L/M ) >/= 0.5 were used to examine the potential lordosis-inhibiting effects of the 5- Q13049 receptor antagonist , R(+)-a- ( 2 , 3-dimethoxyphenyl ) -1-[2(4-fluoro-phenylethyl)]-4-piperidine-methanol ( MDL 100,907 ) , and the P28335 receptor antagonist , 5-methyl-1-(3-pyridylcarbamoyl)-1,2,3,5-tetrahydropyrrolo [ 2 , 3-f ] indole ( SB 206553 ) . Rats with low sexual receptivity ( L/M < 0.5 ) were bilaterally infused with the 5- Q13049 /2C receptor agonist , (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl ( DOI ) , or DOI plus either MDL 100,907 or SB 206553 to determine if either drug would attenuate the lordosis-facilitating effects of DOI . The P28335 receptor antagonist , but not the 5- Q13049 receptor antagonist , effectively inhibited lordosis behavior . Similarly , SB 206553 was more effective than MDL 100,907 in reducing the DOI-induced increase in lordosis responding . However , both drugs limited the duration of lordosis responding initiated by DOI . These results are consistent with prior suggestions that 5- Q13049 /2C receptors within the VMN are involved in the modulation of lordosis behavior and lead to the suggestion that P28335 , rather than 5- Q13049 , receptors are primarily responsible for the effects of 5-HT2 receptor-active drugs on lordosis behavior . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Randomised phase II study of siltuximab ( CNTO 328 ) , an anti- P05231 monoclonal antibody , in combination with mitoxantrone/prednisone versus mitoxantrone/prednisone alone in metastatic castration-resistant prostate cancer . PURPOSE : This open-label phase II trial assessed mitoxantrone/prednisone ( M/P ) with and without siltuximab ( CNTO 328 ) , an anti-interleukin-6 chimeric monoclonal antibody , for patients with metastatic castration-resistant prostate cancer who received prior docetaxel-based chemotherapy . METHODS : Part 1 assessed the safety of biweekly siltuximab 6 mg/kg plus M 12 mg/m(2) every 3 weeks and P. Part 2 assessed efficacy and safety of siltuximab plus M/P versus M/P alone . The primary end-point was progression-free survival ( PFS ) . Progression was defined as progressive disease per Response Evaluation Criteria in Solid Tumours ( RECIST ) , or ≥ 3 new skeletal lesions with clinical deterioration or without deterioration confirmed by repeated bone scan . Rising prostate-specific antigen was not considered progression . RESULTS : DB09036 plus M/P was well tolerated in Part 1 ( n=9 ) . In Part 2 , 48 and 49 patients received siltuximab plus M/P or M/P alone , respectively . Enrolment was prematurely terminated by the Independent Data Monitoring Committee since an apparent imbalance in patient baseline characteristics ( favoring the M/P only arm ) made it unlikely that the study could achieve its primary efficacy end-point . Median PFS was 97 days with siltuximab combination and 228 days with M/P alone ( hazard ratio , 1.72 ; P=0.043 ) . Use of a novel non-validated PFS definition may have contributed to this result . Abnormal laboratory assessments were more frequent with the combination . Infection and febrile neutropenia rates were similar between groups . Greater P02741 suppression was achieved during siltuximab combination treatment compared with M/P alone ( P=0.0003 ) . CONCLUSION : While siltuximab plus M/P appeared well tolerated , improvement in outcomes was not demonstrated . Dose selection of siltuximab for multicentric Castleman 's disease . PURPOSE : DB09036 is a monoclonal antibody that binds to interleukin ( IL ) -6 with high affinity and specificity ; P02741 ( CRP ) is an acute-phase protein induced by P05231 . CRP suppression is an indirect measurement of P05231 activity . Here , modeling and simulation of the pharmacokinetic ( PK ) /pharmacodynamic ( PD ) relationship between siltuximab and CRP were used to support dose selection for multicentric Castleman 's disease ( CD ) . METHODS : PK/PD modeling was applied to explore the relationship between siltuximab PK and CRP suppression following intravenous siltuximab infusion in 47 patients with B cell non-Hodgkin 's lymphoma ( n = 17 ) , multiple myeloma ( n = 13 ) , or CD ( n = 17 ) . DB09036 was administered as 2.8 , 5.5 , or 11 mg/kg q2wks , 11 mg/kg q3wks , or 5.5 mg/kg weekly . Simulations of studied or hypothetical siltuximab dosage regimens ( 15 mg/kg q4wks ) were also performed to evaluate maintenance of CRP suppression below the cutoff value of 1 mg/L . RESULTS : A two-compartment PK model and an inhibitory indirect response PD model adequately described the serum siltuximab and CRP concentration-time profiles simultaneously . PD parameter estimates were physiologically plausible . For all disease types , simulations showed that 11 mg/kg q3wks or 15 mg/kg q4wks would reduce serum CRP to below 1 mg/L after the second dose and throughout the treatment period . CONCLUSIONS : PK/PD modeling was used to select doses for further development of siltuximab in multicentric CD . The dosing recommendation was also supported by the observed efficacy dose-response relationship . CRP suppression in the subsequent randomized multicentric CD study was in agreement with the modeling predictions . Phase 1 study in Japan of siltuximab , an anti- P05231 monoclonal antibody , in relapsed/refractory multiple myeloma . DB09036 , a chimeric monoclonal antibody with high affinity and specificity for interleukin-6 , has been shown to enhance anti-multiple myeloma activity of bortezomib and corticosteroid in vitro . We evaluated the safety , pharmacokinetics , immunogenicity , and antitumor effect of siltuximab in combination with bortezomib and dexamethasone in Japanese patients with relapsed or refractory multiple myeloma . This open-label , phase 1 , dose-escalating study used two doses of siltuximab : 5.5 and 11.0 mg/kg ( administered on day 1 of each 21-day cycle ) . In total , nine patients were treated . The most common grade 3/4 adverse events , lymphopenia ( 89 % ) and thrombocytopenia ( 44 % ) , occurred in patients receiving both doses of siltuximab ; however , no dose-limiting toxicities ( DLTs ) were observed . Following intravenous administration of siltuximab at 5.5 and 11.0 mg/kg , the maximum serum concentration and the area under the curve from 0 to 21 days and from 0 to infinity increased in an approximately dose-proportional manner . Mean half-life , total systemic clearance , and volume of distribution were similar at doses of 5.5 and 11.0 mg/kg . Across both doses , six of the nine patients had complete or partial response ( 22 and 44 % , respectively ) . In conclusion , as no DLT was observed , the recommended dose for this combination is 11.0 mg/kg once every 3 weeks . The study is registered at http://www.clinicaltrials.gov as NCT01309412 .

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Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . Androgen action . Androgens are important for male sex development and physiology . Their actions are mediated by the androgen receptor ( AR ) , a ligand-dependent nuclear transcription factor . The activity of the AR is controlled at multiple stages due to ligand binding and induced structural changes assisted by the foldosome , compartmentalization , recruitment of coregulators , posttranslational modifications and chromatin remodeling , leading to subsequent transcription of androgen-responsive target genes . Beside these short-term androgen actions , there is phenomenological and experimental evidence of long-term androgen programming in mammals and in the human during sensitive programming time windows , both pre- and postnatally . At the molecular level , research into androgen insensitivity syndrome has unmasked androgen programming at the transcriptome level , in genital fibroblasts and peripheral blood mononuclear cells , and at the epigenome level . Androgens are crucial for male sex development and physiology during embryogenesis , at puberty and in adult life . DB00624 and its more potent metabolite , dihydrotestosterone , which is converted from testosterone within the target cell by 5α-reductase II , are the main androgens involved in male sex differentiation . Androgen action is mediated by a single AR . The AR belongs to the nuclear receptor 3 group C , composed of the glucocorticoid receptor ( P04150 ) , mineralocorticoid receptor ( P08235 ) , progesterone receptor ( P06401 ) and AR ( P10275 ) , and acts as a ligand-dependent transcription factor . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Atheroprotective effect of oleoylethanolamide ( OEA ) targeting oxidized LDL . Dietary fat-derived lipid oleoylethanolamide ( OEA ) has shown to modulate lipid metabolism through a peroxisome proliferator-activated receptor-alpha ( Q07869 -α ) -mediated mechanism . In our study , we further demonstrated that OEA , as an atheroprotective agent , modulated the atherosclerotic plaques development . In vitro studies showed that OEA antagonized oxidized LDL ( ox-LDL ) -induced vascular endothelial cell proliferation and vascular smooth muscle cell migration , and suppressed lipopolysaccharide ( LPS ) -induced LDL modification and inflammation . In vivo studies , atherosclerosis animals were established using balloon-aortic denudation ( Q92934 ) rats and ApoE(-/-) mice fed with high-caloric diet ( HCD ) for 17 or 14 weeks respectively , and atherosclerotic plaques were evaluated by oil red staining . The administration of OEA ( 5 mg/kg/day , intraperitoneal injection , i.p. ) prevented or attenuated the formation of atherosclerotic plaques in HCD- Q92934 rats or HCD-ApoE(-/-) mice . Gene expression analysis of vessel tissues from these animals showed that OEA induced the mRNA expressions of Q07869 -α and downregulated the expression of M-CFS , an atherosclerotic marker , and genes involved in oxidation and inflammation , including P35228 , P35354 , P01375 -α and P05231 . Collectively , our results suggested that OEA exerted a pharmacological effect on modulating atherosclerotic plaque formation through the inhibition of LDL modification in vascular system and therefore be a potential candidate for anti-atherosclerosis drug . Gene expression in sexually dimorphic muscles in sheep . DB00624 is known to act differentially on skeletal muscle from different regions of the body . Two genes likely to mediate the testosterone effect are insulin-like growth factor I ( P05019 ) , an important growth regulator acting in an autocrine and paracrine way , and androgen receptor ( AR ) , because receptor density could account for differential muscle growth . Another muscle-specific gene that may play a role in differential muscle growth is myostatin , a member of the transforming growth factor-beta superfamily , shown to be a negative regulator of skeletal muscle mass . The objective of this study was to quantify and compare the steady state expression of these three genes in two different skeletal muscles in sheep . Eleven Dorset rams were slaughtered after reaching puberty and total RNA was extracted from samples of semitendinosus and splenius muscles . P05019 mRNA was measured using a competitive reverse-transcription-polymerase chain reaction . P10275 and myostatin mRNA were measured by a ribonuclease protection assay ( RPA ) with standard curves . The means ( attomoles/microg RNA ) for splenius and semitendinosus muscles were 1.39 and 1.02 ( SE = 0.14 ) , 4.05 and 2.96 ( SE = 0.24 ) , and 4.30 and 3.85 ( SE = 0.37 ) for P05019 , AR , and myostatin , respectively . The difference between the two muscles was significant for P05019 and AR mRNA levels with higher levels in the splenius but not significant for myostatin . Our results show that locally produced P05019 and the regulation of AR expression may be important for sexually dimorphic muscle growth patterns . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . P10275 (CAG)n polymorphism and androgen levels in women with systemic lupus erythematosus and healthy controls . Systemic lupus erythematosus ( SLE ) is an autoimmune disorder that affects mainly females . Therefore , interrelations between the reproductive and immune system have been assumed . Considering the complex influence of hormones and receptors , we aimed to investigate the influence of androgens and androgen receptor ( AR ) polymorphism in women with SLE . One hundred and sixteen patients and 44 healthy women were investigated . DB00624 , sex hormone-binding globulin ( P04278 ) , dehydroepiandrosterone-sulphate ( DHEAS ) concentrations and AR (CAG)n polymorphism were determined . SLE patients had significantly lower levels of total and free testosterone and DHEAS in comparison with the controls . No differences in the CAG repeat length between the groups were established . Women with two alleles carrying more than 22 CAG repeats had significantly higher levels of P04278 ( 101.51 ± 61.81 vs. 69.22 ± 45.93 nmol/l , p = 0.015 ) and DHEAS ( 3.11 ± 2.65 vs. 2.11 ± 3.06 μmol/l , p = 0.007 ) and a tendency to higher testosterone concentrations ( 2.35 ± 2.10 vs. 1.71 ± 1.70 nmol/l , p = 0.056 ) in comparison with other women . The CAG repeat length in the relatively longer (CAG)n allele was inversely related to the Systemic Lupus International Collaborating Clinics/ P10323 index ( r = -0.258 , p = 0.009 ) . In conclusion , the androgen receptor (CAG)n polymorphism is not related to the development of SLE , but it could modulate the severity of the lupus chronic damages as well as the androgen levels in women . P10275 -dependent transactivation of growth arrest-specific gene 6 mediates inhibitory effects of testosterone on vascular calcification . Recent epidemiological studies have found that androgen deficiency is associated with a higher incidence of cardiovascular disease in men . However , little is known about the mechanism underlying the cardioprotective effects of androgens . Here we show the inhibitory effects of testosterone on vascular calcification and a critical role of androgen receptor ( AR ) -dependent transactivation of growth arrest-specific gene 6 ( Gas6 ) , a key regulator of inorganic phosphate ( P(i) ) -induced calcification of vascular smooth muscle cells ( VSMC ) . DB00624 and nonaromatizable androgen dihydrotestosterone inhibited P(i)-induced calcification of human aortic VSMC in a concentration-dependent manner . Androgen inhibited P(i)-induced VSMC apoptosis , an essential process for VSMC calcification . The effects on VSMC calcification were mediated by restoration of P(i)-induced down-regulation of Gas6 expression and a subsequent reduction of Akt phosphorylation . These effects of androgen were blocked by an AR antagonist , flutamide , but not by an estrogen receptor antagonist , DB00947 . We then explored the mechanistic role of the AR in Gas6 expression and found an abundant expression of AR predominantly in the nucleus of VSMC and two consensus ARE sequences in the Gas6 promoter region . DB02901 stimulated Gas6 promoter activity , and this effect was abrogated by flutamide and by AR siRNA . Site-specific mutation revealed that the proximal ARE was essential for androgen-dependent transactivation of Gas6 . Furthermore , chromatin immunoprecipitation assays demonstrated ligand-dependent binding of the AR to the proximal ARE of Gas6 . These results indicate that AR signaling directly regulates Gas6 transcription , which leads to inhibition of vascular calcification , and provides a mechanistic insight into the cardioprotective action of androgens . Regulation of androgen receptor mRNA in rat Sertoli and peritubular cells . Regulation of 9.5-kb androgen receptor mRNA concentrations in Sertoli and peritubular cells from 20-day-old rats was studied by Northern blot analysis . Treatment of cells in vitro for 1-7 days with 300 ng/ml DB00094 increased androgen receptor mRNA up to 4-fold in Sertoli cells but not in peritubular cells . DB00624 ( 100 ng/ml ) had no effect or slightly decreased androgen receptor mRNA in Sertoli and peritubular cells . P10275 mRNA concentrations in Sertoli and peritubular cells from rats killed 15 days after hypophysectomy were elevated 4-5-fold over those in cells from intact rats . The androgen receptor mRNA concentration was decreased in both Sertoli and peritubular cells isolated from hypophysectomized animals treated with 500 micrograms/day testosterone propionate in vivo and subsequently with 100 ng/ml testosterone in vitro . DB00094 treatment ( 100 micrograms/day in vivo , followed by 300 ng/ml in vitro ) did not increase androgen receptor mRNA over that in cells from hypophysectomized controls but rather decreased its concentration to varying degrees in Sertoli and peritubular cells . The rise in androgen receptor mRNA in both Sertoli and peritubular cells isolated from hypophysectomized animals is attributable , at least in part , to the absence of the inhibitory influence of testosterone . Other data in the literature suggest positive regulation of Sertoli cell androgen receptor protein by DB00094 and androgens . Consequently , complex mechanisms involving transcriptional , translational , and post-translational regulation probably control androgen receptor concentrations in the cells of the rat seminiferous tubule . P10275 up-regulates insulin-like growth factor binding protein-5 ( P24593 ) expression in a human prostate cancer xenograft . The insulin-like growth factor ( IGF ) binding proteins ( IGFBPs ) are important modulators of IGF action in many tissues including human prostate . IGFBPs and the androgen receptor ( AR ) are expressed in CWR22 , an androgen-dependent epithelial cell human CaP xenograft that retains biological characteristics of human CaPs , including regression following androgen withdrawal and recurrent growth of AR-containing cells in the absence of testicular androgens beginning several months after castration . Northern blot and in situ hybridization analyses demonstrated that P24593 is androgen-regulated in CWR22 . P24593 messenger RNA ( mRNA ) decreased by 90 % following castration of tumor-bearing mice compared with noncastrate androgen-stimulated mice . DB00624 treatment of CWR22 tumor-bearing mice 6 or 12 days after castration increased P24593 mRNA 10- to 12-fold . Levels of other IGFBP mRNAs did not change following androgen withdrawal and replacement . P24593 protein in tumor extracts bound 125I-labeled P05019 in ligand blot assays and the amounts of P24593 measured by immunoblotting paralleled the levels of P24593 mRNA . Androgen-induced expression of P24593 was at a maximum level within 24 h after testosterone replacement , whereas the major increase in cell proliferation as measured by Ki-67 immunostaining occurred between 24-48 h . This time course suggested P24593 may be a mediator of androgen-induced growth of CWR22 . In tumors that recurred several months following castration , P24593 mRNA and protein increased to levels that approached those in androgen-stimulated CWR22 tumors from noncastrate mice . P24593 immunohistochemical staining of prostate tissue specimens from patients was stronger in androgen-dependent and androgen-independent CaP than in areas of intraepithelial neoplasia ( P63167 ) or benign prostatic hyperplasia ( BPH ) . P24593 mRNA in these specimens was localized predominantly to stromal cells and P24593 protein to epithelial cell membranes . DB06643 -- an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 ) , a cytokine member of the P01375 family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . P01579 -producing ability as a possible marker for the protective T cells against Mycobacterium bovis BCG in mice . We searched for a functional marker with which T cells mediating acquired cellular resistance ( P10323 ) can be discriminated from those mediating delayed-type hypersensitivity ( DTH ) in mice immunized with Mycobacterium bovis BCG . Four wk after injection of the mice with 10(5) viable BCG ( vBCG ) cells emulsified in IFA , a passive transfer experiment revealed that T cells mediating DTH as well as P10323 , which we designated TACR , were generated in the spleen . In contrast , T cells mediating only DTH ( TDTH ) were generated by immunization with 10(7) killed BCG cells along with IFA . The transferring ability of both TACR and TDTH was substantially reduced by treatment with anti-Thy-1.2 or anti- P01730 mAb plus complement , whereas anti-CD8 treatment had no effect . To determine the functional differences between TACR and TDTH , we assessed P60568 and P01579 production from TACR and TDTH after stimulation with PPD in vitro . Similar levels of enhanced P60568 activity were detected in the culture supernatants of both groups of T cells . However , augmented production of P01579 was observed only in TACR . This finding was confirmed by Northern blot analysis for detection of P01579 -specific mRNA . In addition , a significant increase in the number of P01579 -producing cells was observed only in T cells from mice immunized with vBCG . The production of P01579 was also totally abolished by treatment with anti-Thy-1.2 and anti- P01730 mAb plus complement in vitro , whereas anti-CD8 mAb treatment had no effect . These results suggest that P01730 + protective T cells generated by immunization with vBCG are characterized by the ability to produce P01579 after stimulation with specific Ag . Increased dihydrotestosterone receptor levels in high-stage renal adenocarcinoma . Primary renal adenocarcinoma tissue , metastatic deposits , and normal kidney parenchyma from 16 patients were assayed for sex hormone receptors by dextran-coated charcoal adsorption and sucrose gradient centrifugation techniques . DB02901 receptors ( P10275 ) were found in all renal carcinomatous tissue ( 20/20 ) and in 93 % ( 13/14 ) autologous normal kidneys analyzed . DB00624 receptors were found in 84 % ( 16/19 ) of tumors and 93 % ( 14/15 ) or normal kidneys analyzed . Estrogen receptors in small amounts ( ER ) were detected in only 5 % ( 1/19 ) of tumors and in 7 % ( 1/15 ) of normal kidneys . Progesterone receptors ( PR ) in low quantities were detected in 30 % ( 6/20 ) of renal tumors and in 40 % ( 6/15 ) of normal kidneys . P10275 levels in high-stage tumors ( DB00279 , DB00451 ) were significantly elevated over levels in autologous normal kidney , whereas in low-stage tumors localized to the kidney ( T1 and P24752 tumors ) P10275 levels were not significantly different from autologous normal kidney . The mean levels of P10275 in high-stage kidney tumors were significantly elevated over levels in low-stage tumors ( P less than 0.001 ) . P10275 estimation in renal neoplasms may help in biologic staging of renal adenocarcinoma and could define a group of patients in whom anti-androgen therapy may be worth a trial . Bresol inhibits phosphodiesterase 4 gene expression and modulates the levels of select mediators of inflammation in human monocytic cells . Bresol-a poly-herbal formulation , has been reported to be effective against bronchial asthma and allergic rhinitis in children . In vivo studies have supported the anti-histaminic and anti-anaphylactic action of bresol . However , the mechanism of action of bresol in modulation of inflammation has not been studied at the cellular and molecular level . The present study was aimed to elucidate the mechanism(s) of action of bresol at the cellular and molecular levels , using human monocyte leukemia cells . The effects of bresol on phosphodiesterase 4B ( Q07343 ) gene expression were analyzed using human monocytic U937 leukemia cells . The ability of bresol to stimulate DB02527 formation in these cells , as well as its effects on mediators of inflammation like tumor necrosis factor-α ( TNFα ) , nitric oxide ( NO ) , and cycloxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated U937 cells , were also studied . The results here indicated that bresol exhibited potential anti-inflammatory properties by inhibiting LPS-induced Q07343 gene expression in the cells . Bresol also dose dependently activated DB02527 formation , and inhibited TNFα , NO , as well as P35354 formation in the LPS-stimulated cells . Based upon the results , we concluded that the reported anti-inflammatory activity of bresol might be attributed to its abilities to inhibit Q07343 and thus elevate DB02527 levels in human monocytes . The anti-inflammatory effects of bresol might also be a result of the capacity of bresol to modulate the formation of TNFα , NO , and P35354 in monocytes . P10275 is responsible for rat organic cation transporter 2 gene regulation but not for rOCT1 and rOCT3 . PURPOSE : Organic cation transporters 1-3 ( OCT1-3 ; Slc22a1-3 ) mediate the membrane transport of organic cations in the kidney . We previously reported that rat (r)OCT2 expression in the kidney was regulated by testosterone . In this study , we examined the transcriptional mechanisms underlying the testosterone-dependent regulation of rOCT2 expression . METHODS : Approximately 3000-bp fragments of the rOCT1-3 promoter region were isolated , and promoter activities were measured in the renal epithelial cell line LLC- P30613 with the coexpression of rat androgen receptor . RESULTS : Among reporter constructs tested , only rOCT2 promoter activity was stimulated by testosterone . This stimulation was suppressed by nilutamide , an antiandrogen drug . Reporter assays using deletion constructs and mutational constructs of putative androgen response elements ( ARE ) in the rOCT2 promoter region suggested that two AREs , located at approximately -3000 and -1300 , respectively , play an important role in the induction by testosterone . CONCLUSIONS : DB00624 induces the expression of rOCT2 , but not of rOCT1 and rOCT3 , via the AR-mediated transcriptional pathway . This is the first study to address the transcriptional mechanisms of testosterone-dependent gene regulation of the Slc22 family . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . DB00624 and androgen receptor in human nephrolithiasis . PURPOSE : We investigated the relationship of kidney calculi with plasma free and total testosterone , and androgen receptor up-regulation in the kidneys of men with nephrolithiasis . MATERIALS AND METHODS : Male patients with kidney stone and healthy men were included in the study . Blood was collected in a tube containing 2 % heparin in the morning . Total and free serum testosterone was measured by enzyme linked immunosorbent assay . All patients underwent percutaneous nephrostolithotomy . At the end of the procedure ultrasound guided puncture biopsy was done to acquire kidney tissue . Normal kidney tissue obtained at autopsy served as the control . P10275 was detected in kidney tissue by immunohistochemistry . Stone composition was analyzed in each patient . RESULTS : The study included 37 male patients 22 to 39 years old and 31 healthy men 24 to 37 years old . All calculi were composed of calcium oxalate monohydrate or calcium oxalate dihydrate and a few also contained protein or uric acid . Mean±SD serum total and free testosterone was 13.29±4.79 ng/ml and 63.23±28.58 pg/ml in patients , and 7.30±0.82 ng/ml and 35.59±24.91 pg/ml in healthy men , respectively ( each p < 0.001 ) . Immunohistochemistry revealed androgen receptor up-regulation in the kidneys of patients with nephrolithiasis . CONCLUSIONS : Our data suggest the important role of enhanced androgen signaling in human nephrolithiasis . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . D- DB00128 implication in the modulation of frog brain sex steroid levels . There is evidence that D-aspartate ( D- DB00128 ) modulates sex hormone levels in frog testis by regulating the activity of P450 aromatase ( P450 aro ) , the key enzyme which converts DB00624 ( T ) in 17ß-Estradiol ( E2 ) . Here we report , for the first time , that there is a direct correlation among brain levels of D- DB00128 , P450 aro , E2 and Estradiol Receptor ( ERα ) in the male frogs during the reproductive as well as the post-reproductive phases of the breeding cycle , with highest levels being observed in the post-reproductive period . D- DB00128 i.p. administration to frogs ready for reproduction , induced an increase of brain P450 aro protein expression with concomitant enhancement of both E2 levels and ERα expression ; at the same time , brain T levels and P10275 expression decreased . In contrast , in the post-reproductive frogs , D- DB00128 treatment did not modify any of these parameters . Taken together , these results imply that the regulation of P450 aro expression by D- DB00128 could be an important step in the control of E2 levels in the frog brain . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . The androgen receptor : a mediator of diverse responses . Androgens mediate a number of diverse responses through the androgen receptor , a 110 kD ligand-activated nuclear receptor . P10275 expression , which is found in a variety of tissues , changes throughout development , aging , and malignant transformation . The androgen receptor can be activated by two ligands , testosterone and dihydrotestosterone , which bind to the androgen receptor with different affinities . This difference in binding affinity results in different levels of activation of the androgen receptor by the two ligands . The androgen receptor acts as a transcriptional modifier of a variety of genes by binding to an androgen response element . The ability to confer androgen specific actions by the androgen response element may depend on other cell-specific transcription factors and cis-acting DNA elements in close proximity to it . DB00624 and dihydrotestosterone appear to act upon an identical nuclear receptor . However , in certain instances , they mediate different physiologic responses . For example , dihydrotestosterone , but not testosterone , is capable of mediating full sexual development of the male external genitalia . In some cases , the androgen receptor may induce opposite physiologic responses in similar tissue types depending on their location . For example , in male pattern baldness , activated androgen receptors may suppress the growth of distinct hair follicle populations through initiating stromal-epithelial actions , whereas other hair follicles continue to proliferate . In other cases , altered androgen receptor activity due to its mutation or altered expression may lead to pathology such as recurrence of prostate cancer due to development of androgen independence allowing tumor cell proliferation under androgen deprivation . Novel function of androgen receptor-associated protein 55/ O43294 as a negative regulator of P84022 signaling . P10275 -associated protein 55 ( O43294 / O43294 ) belongs to the LIM protein superfamily and is featured by three or four N-terminal LD motifs and four C-terminal zinc finger-like LIM domains . Both LD motifs and LIM domains can serve as protein-protein interaction interfaces . Recently , we found that enforced expression of O43294 inhibits transforming growth factor-beta-mediated up-regulation of Smad binding element-luciferase reporter activity in NRP-154 and NRP-152 rat prostate and LNCaP human prostate cell lines . Moreover , O43294 also inhibits the induction of Smad-binding element 4-luciferase and 3TP-luciferase ( a plasminogen activator inhibitor-1 ( P05121 ) promoter construct ) reporters by constitutively active ( CA ) - P84022 in these cell lines . Co-immunoprecipitation studies suggest an interaction between O43294 and either CA- P84022 or wild-type P84022 in HEK293 cells that occurs through the MH2 domain of P84022 and the C terminus of O43294 with wild-type P84022 having stronger affinity than CA- P84022 to O43294 . O60760 pull-down assays demonstrate that this interaction can occur in a cell-free system . These results are consistent with the luciferase data showing that the C terminus of O43294 is critical for suppression of P84022 activity . Furthermore , using a mammalian two-hybrid system , we confirmed that O43294 interacts with the MH2 domain of P84022 and suppresses CA- P84022 -induced transcriptional responses . In conclusion , these results support that O43294 selectively intercepts transforming growth factor-beta signaling through an interaction of the LIM domain of O43294 with the MH2 domain of P84022 . Sex steroid regulation of chin-marking behavior in male New Zealand rabbits . Chin-marking behavior ( chinning ) was evaluated daily in nine intact adult male rabbits . All subjects ( Ss ) displayed chinning ( mean of means +/- SE = 61 +/- 7 marks/10 min ) but the frequency of this behavior varied largely across them ( range of mean chinning frequency = 19-84 marks/10 min ) . Chinning frequency showed abrupt variations at intervals of 2-3 days , but periodogram analysis did not reveal the existence of an endogenous rhythm in this behavior . Castration significantly decreased ( mean of means +/- SE = 29 +/- 9 marks/10 min ; p < 0.01 ) . but did not suppress chinning . DB00624 propionate ( TP ; 1 mg/day for 16 days ) restored chinning in castrated Ss to slightly below precastration levels ( mean +/- S.E. V 53 +/- 13 marks/10 min ) . The daily administration of 1 microgram estradiol benzoate ( EB ) plus 1 mg dihydrotestosterone propionate ( DHTP ) stimulated chinning within 2 days ( mean increase = 147 % ; p < 0.005 ) . DHTP ( 1 mg/day ) given alone stimulated chinning only after 11 days of treatment ( mean increase = 475 % ; p < 0.01 ) . At higher doses , both DHTP ( 10 mg/day ) and EB ( 10 or 50 micrograms/day ) stimulated chinning by 450 % , 80 % , and 100 % , respectively , over baseline values . Results indicate that chinning largely depends on testicular steroids . P10275 occupation by T or DB02901 , which is enhanced by E , optimally activates chinning . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . DB00624 modulation of dendritic spines of somatosensory cortical pyramidal neurons . Brain structures and functions are increasingly recognized to be directly affected by gonadal hormones , which classically determine reproductive functions and sexual phenotypes . In this regard , we found recently that ovariectomy trimmed the dendritic spines of female rat primary somatosensory cortical neurons and estradiol supplement reversed it . Here , we investigated whether in the male androgen also has a cortical modulatory effect . The dendritic arbors and spines of rat somatosensory cortical pyramidal neurons were studied following intracellular dye injection and three-dimensional reconstruction . Dendritic spines , but not length , of the layers III and V pyramidal neurons were found reduced at 2 weeks and rebounded slightly at 4 weeks and further at 8 and 24 weeks following castration , which , however , remained significantly fewer than those of the intact animals . Two weeks of osmotic pump-delivered testosterone treatment to animals castrated for 4 weeks replenished serum testosterone and reversed the densities of dendritic spines on these neurons to control animal levels . P10275 appears to mediate this effect as its antagonist flutamide reduced the dendritic spines of normal adult rats while causing a mild feedback surge of serum testosterone . On the other hand , blocking the conversion of testosterone to estrogen with the aromatase inhibitor anastrozole failed to alter the dendritic spine densities in male adult rats . In conclusion , these results support our hypothesis that testosterone acts directly on the androgen receptor in males to modulate the dendritic spines of somatosensory cortical output neurons . Anti-allergic function and regulatory mechanisms of KR62980 in allergen-induced airway inflammation . The ligand-activated transcription factor , peroxisome proliferator-activated receptor ( Q07869 )gamma , and its ligands inhibit pro-inflammatory cytokine production by immune cells , thus exerting anti-inflammatory activity . As a non-thiazolidinedione PPARgamma ligand , KR62980 has anti-diabetic and anti-adipogenic activities , but its anti-inflammatory function has yet to be characterized . In this study , we investigated the functions and mechanisms of KR62980 in the activation and differentiation of P01730 + T helper ( Th ) cells by comparing its effects with those of a thiazolidinedione PPARgamma ligand , rosiglitazone . KR62980 dose-dependently and significantly suppressed TCR-triggered Th cell proliferation by suppressing P60568 /IL-2Ralpha-mediated signaling . Both KR62980 and rosiglitazone suppressed IFNgamma production in a dose-dependent manner , whereas P05112 gene expression was specifically suppressed by only KR62980 . In addition , sustained KR62980 treatment diminished Th2 cytokine production by inhibiting c-Maf expression . In vivo administration of KR62980 in a model of allergic asthma significantly attenuated eotaxin-induced eosinophil infiltration , allergic cytokine production and collagen deposition in the lung . KR62980 also decreased goblet cell hyperplasia in the airway and mucous cell metaplasia in nasal epithelium , concurrent with decreases of allergic Th2 cytokines and Q16552 in the draining lymph node . In conclusion , a novel PPARgamma ligand , KR62980 , suppresses in vitro Th2 cell differentiation and attenuates in vivo OVA-induced airway inflammation , suggesting a beneficial role for KR62980 in the treatment of allergic asthma and allergic rhinitis . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Recent androgen receptor antagonists in prostate cancer . P10275 has been shown to promote prostate cell growth and carcinogenesis of prostate cancer by up-regulating its target genes . DB00624 and dihydrotestosterone are two major hormones which bind to and activate androgen receptor . Targeting both the androgen receptor and the enzymes catalyzing the biosynthesis of testosterone and dihydrotestosterone has been shown to be clinically beneficial in the treatment of prostate cancer . Prostate cancer can become castration-resistant after long term treatment with chemo drugs , so efforts in finding compounds with improved efficiency to castration-resistant prostate cancer are urgently needed . In this review we summarized the studies on recent progress in the development of small molecular AR antagonists for the treatment of prostate cancer . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . The effects of the overexpression of recombinant uncoupling protein 2 on proliferation , migration and plasminogen activator inhibitor 1 expression in human vascular smooth muscle cells . AIMS/HYPOTHESIS : Increased oxidative stress in vascular smooth muscle cells ( VSMCs ) has been implicated in the pathogenesis of accelerated atherosclerosis in patients with diabetes mellitus . Uncoupling protein 2 ( P25874 -2 ) is an important regulator of intracellular reactive oxygen species ( ROS ) production . We hypothesised that P25874 -2 functions as an inhibitor of the atherosclerotic process in VSMCs . METHODS : Overexpression of human P25874 -2 was performed in primary cultured human VSMCs ( HVSMCs ) via adenovirus-mediated gene transfer . Its effects on ROS production , AP-1 activity , plasminogen activator inhibitor 1 ( P05121 ) gene expression , and cellular proliferation and migration were measured in response to high glucose and angiotensin II ( Ang II ) concentrations , two major factors in the pathogenesis of atherosclerosis in patients with diabetes and hypertension . Mitochondrial membrane potential and NAD(P)H oxidase activity were also measured . RESULTS : High glucose and Ang II caused transient mitochondrial membrane hyperpolarisation . They also significantly stimulated ROS production , NAD(P)H oxidase activity , mitochondrial membrane potential , AP-1 activity , P05121 mRNA expression , and proliferation and migration of HVSMCs . Adenovirus-mediated transfer of the P25874 -2 gene reversed all of these effects . CONCLUSIONS/INTERPRETATION : The present study demonstrates that P25874 -2 can modify atherosclerotic processes in HVSMCs in response to high glucose and Ang II . Our data suggest that agents increasing P25874 -2 expression in vascular cells may help prevent the development and progression of atherosclerosis in patients with diabetes and hypertension . The impact of biological agents interfering with receptor/ligand binding in the immune system . We herein discuss the impact of biological agents based on the ability of monoclonal antibodies to target specific molecules . This approach has given to clinical immunologists a spectrum of drugs able to manipulate the immune system . In the first session , we discuss drugs targeting T-cell function by : ( 1 ) targeting P10747 mediated costimulation ( DB01281 and DB06681 ) ; ( 2 ) interfering with interleukin-2 receptor ( DB00074 and DB00111 ) ; ( 3 ) blocking cell adhesion and homing ( DB00092 , DB00095 , DB00108 ) . The second session is dedicated to drugs targeting cytokines or their receptors . The best known and largely experimented case is represented by drugs targeting tumor necrosis factor ( P01375 ) ( DB00065 , Adalilumab , Certolizumab ) or its p75 receptor ( DB00005 ) . However , newer products are now available to target other inflammatory cytokines including P05231 , P10145 , IL-12 , P40933 , Q14116 , IL-23 . These agents have the potential to become powerful tools in the control of several immune-mediated diseases , especially auto-immune and inflammatory ones . They traslate into reality the prediction that antibodies will eventually become " magic bullets which seek their own target " ( P. Ehrich , 1906 ) . P10275 in nuclei of rat testis . Testis nuclei of hypophysectomized rats selectively accumulate labeled testosterone and 5alpha-dihydrotestosterone following the injection of tritiated testosterone in vivo . DB00624 and 5alpha-dihydrotestosterone are bound to macromolecules in nuclei and can be extracted with 0.5 M DB00761 . Accumulation of protein bound radioactive androgens in nuclei of isolated seminiferous tubules is similar to that of whole testis . The relative amounts of testosterone and dihydrotestosterone in purified nuclei were similar to the relative amounts bound to cytoplasmic receptors , suggesting that cytoplasmic androgen-receptor complexes may be transported into the nuclei . Binding of labeled androgen is saturable and inhibited by prior injection of unlabeled testosterone or cyproterone acetate . Nuclear binding sites are destroyed by the proteolytic enzyme pronase , but not by DNase . Like the cytoplasmic androgen-receptor complexes in rat testis , nuclear androgen-protein complexes are heat labile and dissociate slowly at 0 degrees C. androgens fail to accumulate in testis nuclei of the Stanley-Gumbreck androgen insensitive rat , a species lacking cytoplasmic androgen receptors in testis and other androgen target tissues . Pubertal maturation is associated with an increase in the number of androgen receptor-immunoreactive cells in the brains of male ferrets . P10275 -immunoreactive ( AR-IR ) cells were identified in brains of male ferrets before and after the onset of pubertal maturation . There was a greater number of AR-IR cells after the onset of pubertal maturation in some , but not all , brain regions examined . Regions in which the number of AR-IR cells increased included the preoptic area and the amygdala , areas known to be involved in the control of male reproductive behaviors . The mechanisms responsible for the increase in AR-IR cells are unknown , but might be related to the higher circulating levels of testosterone that were present in the older animals . DB00624 may increase androgen receptor ( AR ) immunoreactivity both by concentrating already existing ARs within the nucleus and by stimulating de novo synthesis of receptor protein . Chemical ablation of androgen receptor in prostate cancer cells by the histone deacetylase inhibitor LAQ824 . P10275 plays a critical role in the development of primary as well as advanced hormone-refractory prostate cancer . Therefore , ablation of androgen receptor from prostate cancer cells is an interesting concept for developing a new therapy not only for androgen-dependent prostate cancer but also for metastatic hormone-refractory prostate cancer , for which there is no effective treatment available . We report here that LAQ824 , a cinnamyl hydroxamatic acid histone deacetylase inhibitor currently in human clinical trials , effectively depleted androgen receptor in prostate cancer cells at nanomolar concentrations . LAQ824 seemed capable of depleting both the mutant and wild-type androgen receptors in either androgen-dependent and androgen-independent prostate cancer cells . Although LAQ824 may exert its effect through multiple mechanisms , several lines of evidence suggest that inactivation of the heat shock protein-90 ( Hsp90 ) molecular chaperone is involved in LAQ824-induced androgen receptor depletion . Besides androgen receptor , LAQ824 reduced the level of Hsp90 client proteins HER-2 ( ErbB2 ) , Akt/ P31749 , and P04049 in LNCaP cells . Another Hsp90 inhibitor , 17-allyamino-17-demethoxygeldanamycin ( 17- P29372 ) , also induced androgen receptor diminution . LAQ824 induced Hsp90 acetylation in LNCaP cells , which resulted in inhibition of its DB00171 -binding activity , dissociation of Hsp90-androgen receptor complex , and proteasome-mediated degradation of androgen receptor . Consequently , LAQ824 blocked androgen-induced prostate-specific antigen production in LNCaP cells . LAQ824 effectively inhibited cell proliferation and induced apoptosis of these prostate cancer cells . These results reveal that LAQ824 is a potent agent for depletion of androgen receptor and a potential new drug for prostate cancer . Hepatic osteodystrophy : does the osteoprotegerin/receptor activator of nuclear factor-kB ligand system play a role ? Multiple factors can contribute to the development of osteodystrophy in patients with chronic liver disease ( CLD ) . Recently , two new cytokines , osteoprotegerin ( O00300 ) and the receptor activator of nuclear factor-kB ligand ( O14788 ) , have been implicated in the pathogenesis of postmenopausal osteoporosis and other metabolic bone diseases . Therefore , the aim of our study was to evaluate bone metabolism , bone mineral density ( BMD ) and O00300 / O14788 system in 65 male patients with CLD and in 65 healthy controls . Our patients showed lower BMD values than controls both at lumbar and femoral levels . Moreover , they had an unbalanced bone turnover with an increased resorption phase , as shown by high levels of urinary deoxypyridinoline and a decreased formation phase , as shown by the slightly , but significant , low levels of bone-alkaline phosphatase . Patients showed lower plasma levels of free-testosterone than controls and higher - although not significantly so - plasma levels of 17 beta-estradiol . Furthermore , patients with CLD had higher levels of sex hormone-binding globulin and O00300 , and lower levels of 25-hydroxyvitamin D ( 25- Q9BPY8 ) and P05019 than the control group , while O14788 levels were similar in the two groups . In conclusion , our data do not confirm the hypothesis that the O00300 / O14788 system could exert a key role in the pathogenesis of hepatic osteodystrophy , but rather that the observed increase in O00300 levels may represent either the result of the inflammatory process per se or a compensation for the observed enhanced bone resorption . Overexpression of the human 2-oxoglutarate carrier lowers mitochondrial membrane potential in P29320 -293 cells : contrast with the unique cold-induced mitochondrial carrier Q9BZJ4 . Using differential mRNA expression analysis , a previously uncharacterized gene was found to be up-regulated 2-fold in brown adipose tissue ( Q14032 ) of mice exposed to cold ( 4 degrees C ) for 48 h . Contig and homology analysis revealed that the gene represents the murine orthologue to a sequence from a public database encoding a putative human protein ( Q9BZJ4 ) . The presence of mitochondrial carrier domains in the human protein , its transmembrane topology and cold-induction of the mouse Q9BZJ4 gene in Q14032 prompted an analysis of the idea that Q9BZJ4 may represent a new uncoupling protein ( P25874 ) functional homologue . However , transfection of human Q9BZJ4 isoforms in P29320 -293 cells yielded no change in mitochondrial membrane potential ( Deltapsi(m) ) , despite localization of FLAG-tagged Q9BZJ4 to mitochondria of MCF7 cells . Surprisingly , overexpression of the human 2-oxoglutarate carrier ( OGC ) protein ( originally designed as a negative control ) sparked a significant drop in Deltapsi(m) , possibly signalling a previously unappreciated uncoupling activity for the OGC . Androgens and metabolic syndrome : lessons from androgen receptor knock out ( ARKO ) mice . DB00624 ( T ) is an important factor for determining body composition in males . Abdominal obesity is inversely correlated with serum T levels in men , leading to greater mortality . Pathologically hypogonadal men also have a significantly higher fat mass , which is reversed by T administration . However , the mechanism for such anti-obesity effect of androgen has not been well clarified . P10275 ( AR ) null male mice revealed late-onset obesity . Male ARKO mice were euphagic compared to the wild-type male controls , but also less dynamic and less oxygen consuming . Transcript profiling indicated that male ARKO mice had lower transcripts for the thermogenetic uncoupling protein 1 ( P25874 ) . We also found enhanced secretion of adiponectin , which is insulin-sensitizing , from adipose tissue in comparison to wild type , which might partly explain why the overall insulin sensitivity of male ARKO mice remained almost intact despite their apparent obesity . In addition , decreased lipolysis rather than increased lipid synthesis was observed , which might also account for the increased adiposity in male ARKO mice . The results revealed that AR plays important roles in male metabolism by affecting the energy balance , and is negative to both adiposity and insulin sensitivity . P19957 -kinase is essential for ADP-stimulated integrin alpha(IIb)beta3-mediated platelet calcium oscillation , implications for P2Y receptor pathways in integrin alpha(IIb)beta3-initiated signaling cross-talks . Phosphatidylinositol 3-kinase ( PI3K ) pathway is important for platelet activation . Recent studies showed that PI3K and oscillative calcium could cross talk to each other and positively regulate integrin alpha (IIb)beta3-mediated outside-in signaling . However , the mechanism of this feedback regulation remains to be further characterized . Here we found that treatments of both PI3K inhibitor wortmannin and P47900 inhibitor A3P5P could inhibit granular secretion in platelets . Additionally , when RGD-substrate adherent platelets were treated with the ADP scavenger apyrase to deplete the granular-released ADP , their attachments in engaging with substrates became looser and the frequency of calcium oscillation decreased . Since it is known that ADP stimulates the PI3K and calcium signal primarily through Q9H244 and P47900 receptors respectively , our data indicated that integrin alpha(IIb)beta3 downstream PI3K and calcium activation might be not completely coupled to integrin associated signaling complex , but in part through feedback stimulation by granular released ADP . Our data indicates the important roles of PI3K and granular released ADP in coordinating the feedback regulations in integrin alpha(IIb)beta3-mediated platelet activation . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men . DB00624 potentiates the hypoxic ventilatory response of adult male rats subjected to neonatal stress . Neonatal stress disrupts development of homeostatic systems . During adulthood , male rats subjected to neonatal maternal separation ( Q5H8A3 ) are hypertensive and show a larger hypoxic ventilatory response ( HVR ) , with greater respiratory instability during sleep . Neonatal stress also affects sex hormone secretion ; hypoxia increases circulating testosterone of Q5H8A3 ( but not control ) male rats . Given that these effects of Q5H8A3 are not observed in females , we tested the hypothesis that testosterone elevation is necessary for the stress-related increase of the HVR in adult male rats . Pups subjected to Q5H8A3 were placed in an incubator for 3 h per day from postnatal day 3 to 12 . Control pups remained undisturbed . Rats were reared until adulthood , and the HVR was measured by plethysmography ( fractional inspired O2 = 0.12 , for 20 min ) . We used gonadectomy to evaluate the effects of reducing testosterone on the HVR . Gonadectomy had no effect on the HVR of control animals but reduced that of Q5H8A3 animals below control levels . Immunohistochemistry was used to quantify androgen receptors in brainstem areas involved in the HVR . P10275 expression was generally greater in Q5H8A3 rats than in control rats ; the most significant increase was noted in the caudal region of the nucleus tractus solitarii . We conclude that the abnormal regulation of testosterone is important in stress-related augmentation of the HVR . The greater number of androgen receptors within the brainstem may explain why Q5H8A3 rats are more sensitive to testosterone withdrawal . Based on the similarities of the cardiorespiratory phenotype of Q5H8A3 rats and patients suffering from sleep-disordered breathing , these results provide new insight into its pathophysiology , especially sex-based differences in its prevalence . Placental expression of insulin-like growth factor-I , fibroblast growth factor-basic , and neural cell adhesion molecule in preeclampsia . OBJECTIVE : To investigate placental expression of insulin-like growth factor-I ( P05019 ) , fibroblast growth factor-basic ( FGF-b ) , and neural cell adhesion molecule ( N- P62158 ) in preeclampsia . STUDY DESIGN : An immunohistochemical analysis using P05019 , FGF-b , and N- P62158 antibodies was conducted on 4 % paraformaldehyde-fixed placental tissues of preeclamptic patients ( N = 14 ) and normotensive pregnant subjects ( N = 10 ) . Immunostaining patterns of chorionic villi and amniochorionic membranes were assessed . RESULTS : Significantly increased FGF-b and N- P62158 immunoreactivities in cytotrophoblasts and increased FGF-b immunoreactivity in capillary endothelium of chorionic villi of preeclamptic subjects were noted . Significantly increased FGF-b and decreased N- P62158 immunoreactivities in extravillous trophoblasts and decidual cells of amniochorionic membranes obtained from preeclamptic subjects were demonstrated . Additionally , a significantly increased P05019 immunoreactivity was shown in decidual cells of preeclamptic cases . CONCLUSION : Investigation of the regional distribution of P05019 , FGF-b , and N- P62158 at the maternal-fetal interface establishes a better understanding of cell-specific altered growth processes , which may be associated with the pathogenesis of preeclampsia . High-resolution mass spectrometry proteomics for the identification of candidate plasma protein biomarkers for chronic obstructive pulmonary disease . Although cigarette smoking is recognized as the most important cause of chronic obstructive pulmonary disease ( P48444 ) , the pathophysiological mechanisms underlying the lung function decline are not well understood . Using off-line strong cation exchange fractionation with RP-LC- P19957 -MS/MS and robust database searching , 1758 tryptic peptides were identified in plasma samples from cigarette smokers . Using two statistical approaches , 30 peptides were identified to be associated with the annualized rate of lung function decline over 5 years among smokers with P48444 characterized as having rapid ( n = 18 ) or slow ( n = 18 ) decline and 18 smokers without P48444 . The identified peptides belong to proteins that are involved in the complement or coagulation systems or have antiprotease or metabolic functions . This research demonstrates the utility of proteomic profiling to improve the understanding of molecular mechanisms involved in cigarette smoking-related P48444 by identifying plasma proteins that correlate with decline in lung function . DB00624 stimulates proliferation and inhibits interleukin-6 production of normal and hereditary gingival fibromatosis fibroblasts . Hereditary gingival fibromatosis ( P14210 ) is a rare oral condition characterized by a slow and progressive enlargement of the gingiva , involving both the maxilla and mandible . In vitro , P14210 fibroblasts demonstrate a proliferative index significantly higher than fibroblasts from normal gingiva ( NG ) . The objective of this study was to determine the effect of dihydrotestosterone on the proliferation of gingival fibroblasts derived from patients with P14210 ( n = 4 ) and from four healthy individuals . Additionally , we analyzed the effect of dihydrotestosterone on interleukin-6 ( P05231 ) production and determined the expression levels of androgen receptors in NG and P14210 fibroblasts . Gingival fibroblasts from NG and P14210 were incubated with increasing concentrations of dihydrotestosterone with or without androgen blockers , and cultured for 24 h , and the proliferation index was determined by automated cell counter . P05231 production , in this system , was quantified using a " capture " enzyme-linked immunosorbent assay ( ELISA ) . Semi-quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR ) was performed to measure the mRNA expression of androgen receptors . The results indicated that dihydrotestosterone simultaneously downregulates the production of P05231 and upregulates the cell proliferation . DB01216 and cyprosterone acetate , two anti-androgens , partially reversed these effects . P10275 mRNA expression was identified in both NG and P14210 fibroblasts ; however , the levels in NG were higher than those observed in P14210 . These results show that testosterone coordinates the proliferation and production of P05231 of normal and P14210 fibroblasts . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . P50591 -deficiency accelerates vascular calcification in atherosclerosis via modulation of O14788 . The osteoprotegerin ( O00300 ) and receptor activator of nuclear factor-κB ligand ( O14788 ) cytokine system , not only controls bone homeostasis , but has been implicated in regulating vascular calcification . P50591 ( P50591 ) is a second ligand for O00300 , and although its effect in vascular calcification in vitro is controversial , its role in vivo is not yet established . This study aimed to investigate the role of P50591 in vascular calcification in vitro using vascular smooth muscle cells ( VSMCs ) isolated from P50591 (-/-) and wild-type mice , as well as in vivo , in advanced atherosclerotic lesions of P50591 (-/-)ApoE(-/-) mice . The involvement of O00300 and O14788 in this process was also examined . P50591 dose-dependently inhibited calcium-induced calcification of human VSMCs , while P50591 (-/-) VSMCs demonstrated accelerated calcification induced by multiple concentrations of calcium compared to wild-type cells . Consistent with this , O14788 mRNA was significantly elevated with 24 h calcium treatment , while O00300 and P50591 expression in human VSMCs was inhibited . Brachiocephalic arteries from P50591 (-/-)ApoE(-/-) and ApoE(-/-) mice fed a high fat diet for 12 w demonstrated increased chondrocyte-like cells in atherosclerotic plaque , as well as increased aortic collagen II mRNA expression in P50591 (-/-)ApoE(-/-) mice , with significant increases in calcification observed at 20 w . P50591 (-/-)ApoE(-/-) aortas also had significantly elevated O14788 , P12643 , IL-1β , and Q07869 -γ expression at 12 w . Our data provides the first evidence that P50591 deficiency results in accelerated cartilaginous metaplasia and calcification in atherosclerosis , and that P50591 plays an important role in the regulation of O14788 and inflammatory markers mediating bone turn over in the vasculature . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Serum cytokine level and production of reactive oxygen species ( ROS ) by blood neutrophils from a schizophrenic patient with hypersensitivity to neuroleptics . BACKGROUND : There have been few publications concerning the role of the immune system in neuroleptic intolerance . Some studies have shown that in neuroleptic malignant syndrome ( Q5H8A3 ) , associated with disseminated intravascular coagulation ( DIC ) , the serum level of tumor necrosis factor alpha ( P01375 ) increases significantly , which is thought to trigger the onset of DIC . CASE REPORT : A 23-year-old woman suffering from catatonic schizophrenia developed hypersensitivity to neuroleptics . One month before being referred to the present authors , she had a haloperidol-induced Q5H8A3 episode in another psychiatric hospital , with high temperature , CPK activity , muscle rigidity and leukocytosis . On admission to our clinic and after treatment with promazine , laboratory tests showed an increase in serum CPK activity and mild leukocytosis . Neuroleptic treatment was discontinued , and the serum level of CPK and white blood cell count was monitored daily for 7 days , as well as the serum level of some cytokines and the production of reactive oxygen species ( ROS ) by blood neutrophils . The serum levels of interleukin 1 alpha ( IL-1 ) , P05231 and P01375 changed significantly over the observation period , forming waves with peak activity of P05231 and P01375 exceeding normal levels . The level of P01583 was within the control range . ROS production by the patient 's blood neutrophils was also increased , as well as catalase serum activity . CONCLUSIONS : Some proinflammatory cytokines may participate in the mechanisms leading to the development of neuroleptic intolerance in schizophrenic patients . Cytokine-stimulated ROS production may participate in tissue injury and increase CPK serum activity . Effect of interferon-gamma and P01375 on P15941 mucin expression in ovarian carcinoma cell lines . In view of the potential uses of cell surface tumour associated antigens in novel anticancer treatment , a study was designed to investigate whether the biological response modifiers interferon-gamma ( P01579 ) and tumour necrosis factor-alpha ( P01375 ) could effect the expression of an epitope on the tumour associated P15941 epithelial mucin . Four ovarian carcinoma cell lines showing high ( OAW42 and GG ) and low ( JAM and PE01 ) basal expression of P15941 were treated with 10-1000 U/mL of P01579 or P01375 for one or five days . Changes in P15941 expression in cells exposed to P01579 or P01375 were monitored using an ELISA technique with the monoclonal antibody O43633 which reacts with a core protein epitope on the P15941 mucin , and then corrected for the number of viable cells present . P01375 had little effect on P15941 expression , but one or five days exposure to P01579 significantly increased P15941 expression ( p < 0.01 ) in all cell lines including the two cell lines that initially showed little or no expression . Serum testosterone plays an important role in the metastatic ability of castration resistant prostate cancer . PURPOSE : Prostate cells are dependent on androgens for growth and proliferation . Androgen deprivation therapy is the recommended treatment for advanced/metastatic prostate cancer . Under this therapy , prostate cancer will inevitably progress to castration resistant prostate cancer ( CRPC ) . Despite putative castration resistance , testosterone might still play a crucial role in the progression of CRPC . The goal of this study was to determine the role of testosterone in the formation of metastases of CRPC in both in vitro and in vivo settings . METHODS : In vitro , the effect of testosterone and the non-aromatizable androgen methyltrienolone on migration , invasion and proliferation of a castration-resistant prostate cancer rat cell line ( Dunning R3327-MATLyLu ) was assessed using a transwell assay and a sulforhodamine B assay and immunohistochemical detection of ki67 . P10275 status was determined using Western blot . In vivo , Copenhagen rats were divided in four groups ( males , females , castrated males and females with testosterone suppletion ) and inoculated with MATLyLu cells . Tumor size was assessed daily . RESULTS : DB00624 increased cell migration and invasion in a concentration-dependent manner in vitro . DB00624 did not affect in vitro cell proliferation . No difference was shown between the effect of testosterone and methyltrienolone . In vivo , in groups with higher levels of circulating testosterone , more rats had (micro)metastases compared with groups with low levels of testosterone . No effect was observed on primary tumor size/growth . CONCLUSIONS : Despite assumed castration resistance , progression of prostate cancer is still influenced by androgens . Therefore , continuous suppression of serum testosterone in patients who show disease progression during castration therapy is still warranted . [ Hemostatic evaluation of a patient with haloperidol-induced neuroleptic malignant syndrome associated with disseminated intravascular coagulation ] . A 94-year-old man who had been admitted to our hospital for the treatment of senile dementia and restless behavior exhibited consciousness disturbances , acute respiratory failure , high fever , and thrombocytopenia the day after receiving haloperidol as prescribed by a psychiatrist . On the fourth day following administration of haloperidol , acute renal failure with rhabdomyolysis and disseminated intravascular coagulation ( DIC ) developed in the patient , who was accordingly given a diagnosis of haloperidol-induced neuroleptic malignant syndrome ( Q5H8A3 ) associated with DIC . He was then given heparin and antithrombin III , and his DIC symptoms improved soon thereafter . Elevated plasma levels of tissue factor and tumor necrosis factor-alpha ( P01375 ) were sustained during this therapy course . Other cytokines , including interleukin P01584 , P60568 and P05231 , were not elevated . There are activation of extrinsic coagulation and an elevated level of P01375 during acute renal failure and rhabdomyolysis associated with Q5H8A3 , which is thought to trigger the onset of DIC . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . P10275 CAG repeat polymorphism is associated with serum testosterone levels , obesity and serum leptin in men with type 2 diabetes . OBJECTIVE : To determine the relationships between androgen receptor CAG repeat polymorphism length ( AR CAG ) , sex hormones and clinical variables in men with type 2 diabetes ( DM2 ) . Men with DM2 are known to have a high prevalence of low testosterone levels . Studies suggest that testosterone replacement therapy may improve insulin sensitivity and glycaemic control in men with DM2 and reduces central obesity and serum leptin . AR CAG is known to correlate negatively with AR sensitivity and positively with body fat , insulin levels , and leptin in healthy men . DESIGN : Cross-sectional study set in a district general hospital diabetes centre . METHODS : Sex hormones , AR CAG and symptoms of hypogonadism were assessed in 233 men with DM2 . Associations were sought between these variables and others such as obesity , leptin , glycaemic control , and blood pressure . RESULTS : DB00624 was negatively associated and AR CAG positively associated with obesity and leptin . The associations of AR CAG with leptin and obesity were independent of testosterone , estradiol , gonadotropins , and age . AR CAG was also independently associated with total , bioavailable and free testosterone , LH , waist circumference , body mass index , leptin , and systolic blood pressure . There was no association of AR CAG with sex hormone binding globulin , estradiol , HbA(1C) or the symptoms of hypogonadism . CONCLUSIONS : The association of longer AR CAG with obesity and leptin suggests that shorter AR CAG may have an influence in maintaining healthy anthropomorphics and metabolism in men with DM2 . DB00624 and LH levels are higher in men with longer AR CAG , probably reflecting reduced negative feedback through a less sensitive receptor . P10275 coregulator Q96L73 -alpha interacts with death receptor-6 revealed by the yeast two-hybrid . Q96L73 -alpha is a newly identified androgen receptor coactivator . In order to further elucidate its precise role in cells , using the Q96L73 -alpha fragment containing four P20941 and one Q01105 conserved domains as bait we revealed an Q96L73 -alpha- P20941 - Q01105 -interacting protein , death receptor-6 ( O75509 ) , in the yeast two-hybrid screening . O75509 is the member of P01375 receptor family and has a death domain in its intracellular cytoplasmic portion ( DR6cp ) to mediate the cell apoptosis . The interaction between Q96L73 -alpha- P20941 - Q01105 and DR6cp was confirmed in vitro and in vivo . Our finding implied that androgen signaling pathway might cross talk with apoptosis signaling pathway through the interaction between Q96L73 -alpha and O75509 . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . Q07869 gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 - and P15692 -mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 ) - and basic fibroblast growth factor ( P09038 ) -induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 - and P09038 -induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 and P15692 in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients . Delayed haemolytic transfusion reaction caused by anti-M antibody in a patient receiving interleukin-2 and interferon for metastatic renal cell cancer . Anti-M is usually a naturally occurring cold-reactive immunoglobulin M ( IgM ) antibody , often with an immunoglobulin G ( IgG ) component , and is seldom implicated in delayed haemolytic transfusion reactions ( P10275 ) . However , cases have been reported . In the majority , a P10275 is not suspected until further blood is requested and a new antibody is detected on pretransfusion testing . We describe the case of a young man receiving therapy with interleukin-2 ( P60568 ) and interferon-alpha ( IFN-alpha ) for metastatic renal cell cancer who developed a clinically suspected P10275 that was confirmed serologically to be caused by anti-M , reactive at 37 degrees C . We discuss the possible role of his biochemotherapy in the development of the P10275 .

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interacts_with DB00470?

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Tolerance to chronic DB00470 ( Δ⁹-THC ) in rhesus macaques infected with simian immunodeficiency virus . Although Δ⁹-THC has been approved to treat anorexia and weight loss associated with AIDS , it may also reduce well-being by disrupting complex behavioral processes or enhancing HIV replication . To investigate these possibilities , four groups of male rhesus macaques were trained to respond under an operant acquisition and performance procedure , and administered vehicle or Δ⁹-THC before and after inoculation with simian immunodeficiency virus ( SIV(mac251) , 100 TCID₅₀/ml , i.v. ) . Prior to chronic Δ⁹-THC and SIV inoculation , 0.032-0.32 mg/kg of Δ⁹-THC produced dose-dependent rate-decreasing effects and small , sporadic error-increasing effects in the acquisition and performance components in each subject . Following 28 days of chronic Δ⁹-THC ( 0.32 mg/kg , i.m. ) or vehicle twice daily , DB00470 -treated subjects developed tolerance to the rate-decreasing effects , and this tolerance was maintained during the initial 7-12 months irrespective of SIV infection ( i.e. , +THC/-SIV , +THC/+SIV ) . Q8N1N2 necropsy was performed on all SIV subjects an average of 329 days post-SIV inoculation , with postmortem histopathology suggestive of a reduced frequency of CNS pathology as well as opportunistic infections in DB00470 -treated subjects . Chronic Δ⁹-THC also significantly reduced CB-1 and P34972 receptor levels in the hippocampus , attenuated the expression of a proinflammatory cytokine ( P13500 ) , and did not increase viral load in plasma , cerebrospinal fluid , or brain tissue compared to vehicle-treated subjects with SIV . Together , these data indicate that chronic Δ⁹-THC produces tolerance to its behaviorally disruptive effects on complex tasks while not adversely affecting viral load or other markers of disease progression during the early stages of infection . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Inhibition of P26718 receptor function by antibody therapy attenuates transfer-induced colitis in SCID mice . A role for the activating NK-receptor P26718 has been indicated in several autoimmune diseases in humans and in animal models of type 1 diabetes and multiple sclerosis , and treatment with monoclonal antibodies to P26718 attenuated disease severity in these models . In an adoptive transfer-induced model of colitis , we found a significantly higher frequency of P01730 (+) P26718 (+) cells in blood , mesenteric lymph nodes , colon , and spleen from colitic mice compared to BALB/c donor-mice . We , therefore , wanted to study the effect of anti- P26718 antibody ( P34972 ) treatment initiated either before onset of colitis , when the colitis was mild , or when severe colitis was established . P34972 treatment decreased the detectable levels of cell-surface P26718 and prophylactic administration of P34972 attenuated the development of colitis significantly , whereas a more moderate reduction in the severity of disease was observed after P34972 administration to mildly colitic animals . P34972 did not attenuate severe colitis . We conclude that the frequency of P01730 (+) P26718 (+) cells increase during development of experimental colitis . P26718 may play a role in the early stages of colitis in this model , since early administration of P34972 attenuated disease severity . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Live-cell monitoring of periodic gene expression in synchronous human cells identifies Forkhead genes involved in cell cycle control . We developed a system to monitor periodic luciferase activity from cell cycle-regulated promoters in synchronous cells . Reporters were driven by a minimal human Q01094 promoter with peak expression in P55008 /S or a basal promoter with six Forkhead DNA-binding sites with peak expression at G2/M . After cell cycle synchronization , luciferase activity was measured in live cells at 10-min intervals across three to four synchronous cell cycles , allowing unprecedented resolution of cell cycle-regulated gene expression . We used this assay to screen Forkhead transcription factors for control of periodic gene expression . We confirmed a role for Q08050 and identified two novel cell cycle regulators , Q9UPW0 and FOXK1 . Knockdown of Q9UPW0 and FOXK1 eliminated cell cycle-dependent oscillations and resulted in decreased cell proliferation rates . Analysis of genes regulated by Q9UPW0 and FOXK1 showed that Q9UPW0 may regulate a network of zinc finger proteins and that FOXK1 binds to the promoter and regulates P00374 , P04818 , P57764 , and the E2F binding partner Q14186 . Chromatin immunoprecipitation followed by high-throughput sequencing analysis identified 4329 genomic loci bound by FOXK1 , 83 % of which contained a FOXK1-binding motif . We verified that a subset of these loci are activated by wild-type FOXK1 but not by a FOXK1 ( H355A ) DNA-binding mutant . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . delta 9- DB00470 increases activity of tyrosine hydroxylase in cultured fetal mesencephalic neurons . The exposure of pregnant rats to delta 9-tetrahydrocannabinol ( delta 9-THC ) , the main psychoactive constituent of Cannabis sativa , during gestation and lactation , affects the gene expression and the activity of tyrosine hydroxylase ( TH ) in the brain of their offspring , measured at fetal and early postnatal ages , when the expression of this enzyme plays an important role in neural development . In the present article , we have examined whether delta 9-THC is able to affect TH activity in cultured mesencephalic neurons obtained from fetuses at gestational d 14 . Thus , TH activity increased approximately twofold in cells obtained from naive fetuses when exposed for 24 h to medium containing delta 9-THC . In addition , TH activity was also approx twofold higher in cells obtained from fetuses exposed daily to delta 9-THC from d 5 of gestation than in cells obtained from control fetuses , when both were exposed to basal media . This effect of delta 9-THC on TH activity seems to be produced via the activation to cannabinoid receptors , in particular the P21554 subtype , which would presumably be located in these cells . This is because the exposure to medium containing both delta 9-THC and SR141716A , a specific antagonist for P21554 receptors , abolished the effect observed with delta 9-THC alone . SR141716A alone was without effect on TH activity . Collectively , our results support the notion that delta 9-THC increased TH activity in cultured mesencephalic neurons , as previously observed in vivo , and that this effect was produced by activation of P21554 receptors , which seem to be operative at these early ages . All this points to a role for the endogenous cannabimimetic system in brain development . Neonatal melanocortin receptor agonist treatment reduces play fighting and promotes adult attachment in prairie voles in a sex-dependent manner . The melanocortin receptor ( P08235 ) system has been studied extensively for its role in feeding and sexual behavior , but effects on social behavior have received little attention . α-MSH interacts with neural systems involved in sociality , including oxytocin , dopamine , and opioid systems . Acute melanotan-II ( MTII ) , an MC3/4R agonist , potentiates brain oxytocin ( OT ) release and facilitates OT-dependent partner preference formation in socially monogamous prairie voles . Here we examined the long-term impact of early-life P08235 stimulation on hypothalamic neuronal activity and social development in prairie voles . Male and female voles were given daily subcutaneous injections of 10 mg/kg MTII or saline between postnatal days ( P01160 ) 1-7 . Neonatally-treated males displayed a reduction in initiated play fighting bouts as juveniles compared to control males . Neonatal exposure to MTII facilitated partner preference formation in adult females , but not males , after a brief cohabitation with an opposite-sex partner . Acute MTII injection elicited a significant burst of the immediate early gene P18146 immunoreactivity in hypothalamic OT , vasopressin , and corticotrophin releasing factor neurons , when tested in P01160 6-7 animals . Daily neonatal treatment with 1 mg/kg of a more selective , brain penetrant P32245 agonist , PF44687 , promoted adult partner preferences in both females and males compared with vehicle controls . Thus , developmental exposure to P08235 agonists lead to a persistent change in social behavior , suggestive of structural or functional changes in the neural circuits involved in the formation of social relationships . Delta(9)- DB00470 enhances MCF-7 cell proliferation via cannabinoid receptor-independent signaling . We recently reported that Delta(9)-tetrahydrocannabinol ( Delta(9)-THC ) has the ability to stimulate the proliferation of human breast carcinoma MCF-7 cells . However , the mechanism of action remains to be clarified . The present study focused on the relationship between receptor expression and the effects of Delta(9)-THC on cell proliferation . RT-PCR analysis demonstrated that there was no detectable expression of CB receptors in MCF-7 cells . In accordance with this , no effects of cannabinoid 1/2 ( P21554 /2 ) receptor antagonists and pertussis toxin on cell proliferation were observed . Although MCF-7 cell proliferation is suggested to be suppressed by Delta(9)-THC in the presence of CB receptors , it was revealed that Delta(9)-THC could exert upregulation of living cells in the absence of the receptors . Interestingly , Delta(9)-THC upregulated human epithelial growth factor receptor type 2 ( P04626 ) expression , which is known to be a predictive factor of human breast cancer and is able to stimulate cancer cells as well as MCF-7 cells . DB00970 -treatment interfered with the upregulation of P04626 and cell proliferation by cannabinoid . Taken together , these studies suggest that , in the absence of CB receptors , Delta(9)-THC can stimulate the proliferation of MCF-7 cells by modulating , at least in part , P04626 transcription . Evidence for an interaction between P21554 cannabinoid and melanocortin P08235 -4 receptors in regulating food intake . P32245 ( MCR4 ) and CB(1) cannabinoid receptors independently modulate food intake . Although an interaction between the cannabinoid and melanocortin systems has been found in recovery from hemorrhagic shock , the interaction between these systems in modulating food intake has not yet been examined . The present study had two primary purposes : 1 ) to examine whether the cannabinoid and melanocortin systems act independently or synergistically in suppressing food intake ; and 2 ) to determine the relative position of the CB(1) receptors in the chain of control of food intake in relation to the melanocortin system . Rats were habituated to the test environment and injection procedure and then received intracerebroventicular injections of various combinations of the MCR4 receptor antagonist JKC-363 , the CB(1) receptor agonist Delta(9)-tetrahydrocannabinol , the MCR4 receptor agonist alpha-MSH , or the cannabinoid CB(1) receptor antagonist SR 141716 . Food intake and locomotor activity were then recorded for 120 min . When administrated alone , SR 141716 and alpha-MSH dose-dependently attenuated baseline feeding , whereas sub-anorectic doses of SR 141716 and alpha-MSH synergistically attenuated baseline feeding when combined . Delta(9)- DB00470 -induced feeding was not blocked by alpha-MSH , whereas SR 141716 dose-dependently attenuated JKC-363-induced feeding . Locomotor activity was not significantly affected by any drug treatment , suggesting that the observed effects on feeding were not due to a nonspecific reduction in motivated behavior . These findings revealed a synergistic interaction between the cannabinoid and melanocortin systems in feeding behavior . These results further suggested that CB(1) receptors are located downstream from melanocortin receptors and CB(1) receptor signaling is necessary to prevent the melanocortin system from altering food intake . Identification of a high-affinity binding site involved in the transport of endocannabinoids . Phytocannabinoids , such as the principal bioactive component of marijuana , delta9-tetrahydrocannabinol , have been used for thousands of years for medical and recreational purposes . DB00470 and endogenous cannabinoids ( e.g. , anandamide ) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors ( P21554 and CB2 ) . The biosynthesis and physiology of anandamide is well understood , but its mechanism of uptake ( resulting in signal termination by fatty acid amide hydrolase ) has been elusive . Mounting evidence points to the existence of a specific anandamide transport protein ; however , no direct evidence for this protein has been provided . Here , we use a potent , competitive small molecule inhibitor of anandamide uptake ( LY2318912 , IC50 7.27 +/- 0.510 nM ) to identify a high-affinity , saturable anandamide transporter binding site ( LY2318912 ; K(d) = 7.62 +/- 1.18 nM , B(max) = 31.6 +/- 1.80 fmol/mg protein ) that is distinct from fatty acid amide hydrolase . Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function . Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling , and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors . The stimulation of ketogenesis by cannabinoids in cultured astrocytes defines carnitine palmitoyltransferase I as a new ceramide-activated enzyme . The effects of cannabinoids on ketogenesis in primary cultures of rat astrocytes were studied . Delta9- DB00470 ( THC ) , the major active component of marijuana , produced a malonyl- DB01992 -independent stimulation of carnitine palmitoyltransferase I ( CPT-I ) and ketogenesis from [14C]palmitate . The THC-induced stimulation of ketogenesis was mimicked by the synthetic cannabinoid HU-210 and was prevented by pertussis toxin and the P21554 cannabinoid receptor antagonist SR141716 . Experiments performed with different cellular modulators indicated that the THC-induced stimulation of ketogenesis was independent of cyclic AMP , Ca2+ , protein kinase C , and mitogen-activated protein kinase ( MAPK ) . The possible involvement of ceramide in the activation of ketogenesis by cannabinoids was subsequently studied . THC produced a P21554 receptor-dependent stimulation of sphingomyelin breakdown that was concomitant to an elevation of intracellular ceramide levels . Addition of exogenous sphingomyelinase to the astrocyte culture medium led to a MAPK-independent activation of ketogenesis that was quantitatively similar and not additive to that exerted by THC . Furthermore , ceramide activated CPT-I in astrocyte mitochondria . Results thus indicate that cannabinoids stimulate ketogenesis in astrocytes by a mechanism that may rely on P21554 receptor activation , sphingomyelin hydrolysis , and ceramide-mediated activation of CPT-I . delta(9)- DB00470 increases nerve growth factor production by prostate PC-3 cells . Involvement of P21554 cannabinoid receptor and P04049 . Cannabinoids , the active components of marihuana , exert a variety of effects in humans . Many of these effects are mediated by binding to two types of cannabinoid receptor , P21554 and CB2 . Although P21554 is located mainly in the central nervous system , it may also be found in peripheral tissues . Here , we study the effect of cannabinoids in the production of nerve growth factor by the prostate tumor cell line PC-3 . We show that addition of Delta(9)-tetrahydrocannabinol to PC-3 cells stimulated nerve growth factor production in a dose-dependent and time-dependent manner . Maximal effect was observed at 0.1 microM Delta(9)-tetrahydrocannabinol and 72 h of treatment . Stimulation was reversed by the P21554 antagonists AM 251 and SR 1411716A . Pre-treatment of cells with pertussis toxin also prevented the effect promoted by Delta(9)-tetrahydrocannabinol . These results indicate that Delta(9)-tetrahydrocannabinol stimulation of nerve growth factor production in these cells was mediated by the cannabinoid P21554 receptor . The implication of P04049 activation in the mode of action of Delta(9)-tetrahydrocannabinol is also suggested . Construction of a steric map of the binding pocket for cannabinoids at the cannabinoid receptor . In order to gain information about the topology of the brain cannabinoid receptor ( P21554 ) , a Receptor Steric ( RS ) Map for cannabinoids at this receptor was calculated . The classical cannabinoids (-)-11-hydroxy- DB00470 ( P04264 = 210 +/- 56 nM ) , (-)-9-nor-9-beta-hydroxy-hexahydrocannabinol ( P04264 = 124 +/- 17 nM ) , nabilone ( P04264 = 120 +/- 13 nM ) , and the non-classical cannabinoid , CP-55,244 ( P04264 = 1.4 +/- .3 nM ) were used as template molecules . The RS map was obtained as the union of the van der Waals ' volumes of only those accessible conformers identified by P08253 calculations that were able to clear a region of steric interference at the P21554 receptor previously characterized by us [ Reggio , P.H. , Panu , A.M. and Miles , S. ( 1993 ) , J. Med. Chem. , 36 , 1761-1771 ] . The utility of the RS Map was explored by screening the accessible conformers of the classical cannabinoid , cannabinol ( CBN ) , ( P04264 = 3200 +/- 450 nM ) , for its ability to fit within the RS map . Only the global minimum energy conformer of CBN ( 53.2 % abundance at 298K ) was able to fit within the RS map . These results imply that one reason for the reduced affinity of CBN may be that only 53.2 % of CBN molecules are shaped properly to fit in the binding pocket for cannabinoids at the P21554 receptor . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . Cannabinoid agonists in the treatment of blepharospasm -- a case report study . The benign essential blepharospasm is a subliminal form of primary torsion dystonia with still uncertain aetiology . It is characterized by involuntary convulsive muscle contractions of the M. orbicularis occuli , accompanied by unbearable pain of the cornea , eye bulb and the muscle itself . It has been suggested that blepharospasm is neurobiologically based on a dysfunction of the basal ganglia and an impairment of the dopamine neurotransmitter system . Therefore , therapy of blepharospasm contains administration of anticholinergic- and tranquillizing drugs as well as botulinum toxin as neuromuscular blocking agent . However serious side effects can be observed as well as failure of therapy . In the brain a dense co-localisation of cannabinoid ( P21554 ) and dopamine ( D2 ) -receptor was identified which had been associated with the influence of cannabinoids on the dopaminergic reward system . Additionally , it has been demonstrated that cannabinoids may have an impact on the central GABAergic and glutaminergic transmitter system and thus might be involved in the influence of movement control . In the present case we administered the cannabinoid receptor agonist Dronabinol ( Delta-9- DB00470 ) to a woman suffering from severe blepharospasm . Multiple treatments with botulinum toxin did not reveal a long-lasting beneficial effect . By contrast , treatment with 25 mg Dronabinol for several weeks improved the patients ' social life and attenuated pain perception remarkably . This case study demonstrates that the therapy with a cannabinoid agonist may provide a novel tool in the treatment of blepharospasm and maybe of other multifactorial related movement disorders . Development of a yeast protein fragment complementation assay ( DB11245 ) system using dihydrofolate reductase ( P00374 ) with specific additives . A yeast protein fragment complementation assay ( DB11245 ) system based on dihydrofolate reductase ( P00374 ) is difficult to be operated because it is not as sensitive to trimethoprim ( P54849 ) as the system using a prokaryotic microorganism . Here , the DB11245 system using P00374 , specific inhibitors , and a substrate in the yeast Saccharomyces cerevisiae was newly developed . As a model , the human oncoprotein Ras and the Ras-binding domain ( RBD ) of P04049 were individually and genetically fused to P00374 fragment , and each genetic construct was coexpressed under the control of the P22466 promoter . An interaction between Ras and RBD could be evaluated on the basis of cell proliferation . To establish the experimental conditions for the yeast DB11245 system based on the P00374 reconstitution , we examined yeast host strains and the concentration of inhibitory additives to prevent endogenous P00374 activity , namely , P54849 and sulfanilamide , and the substrate of P00374 , namely , folic acid . The transformant harboring wild-type Ras or its variants showed positive interaction signals , and the order of interactions for combination corresponded to the results of other in vitro assays . Moreover , combinatorial mutated Ras-binding domains were constructed , and the interaction of RBDs with Ras using this yeast DB11245 system was examined . As a result , various types of mutated clone for RBD were obtained . These demonstrations suggest that the yeast DB11245 system based on P00374 can be one of good , convenient , and inexpensive tools for investigating eukaryotic protein-protein interactions in vivo . Distribution of cannabinoid receptors in the central and peripheral nervous system . P21554 cannabinoid receptors appear to mediate most , if not all of the psychoactive effects of DB00470 and related compounds . This G protein-coupled receptor has a characteristic distribution in the nervous system : It is particularly enriched in cortex , hippocampus , amygdala , basal ganglia outflow tracts , and cerebellum -- a distribution that corresponds to the most prominent behavioral effects of cannabis . In addition , this distribution helps to predict neurological and psychological maladies for which manipulation of the endocannabinoid system might be beneficial . P21554 receptors are primarily expressed on neurons , where most of the receptors are found on axons and synaptic terminals , emphasizing the important role of this receptor in modulating neurotransmission at specific synapses . While our knowledge of P21554 localization in the nervous system has advanced tremendously over the past 15 years , there is still more to learn . Particularly pressing is the need for ( 1 ) detailed anatomical studies of brain regions important in the therapeutic actions of drugs that modify the endocannabinoid system and ( 2 ) the determination of the localization of the enzymes that synthesize , degrade , and transport the endocannabinoids . The Sprouty-related protein , Q7Z699 , localizes in a lipid raft/caveola and inhibits P29323 activation in collaboration with caveolin-1 . Q03135 ( Cav-1 ) has been suggested to function as a negative regulator of mitogen-stimulated proliferation and the Ras- Q8NFH3 /44 P29323 ( Q96HU1 kinase ) pathway in a variety of cell types . However , the molecular basis of this suppression has not been clarified . Spred/Sprouty family proteins are also negative regulators of the P29323 pathway by interacting with P04049 . The Spred/Sprouty family proteins contain a cysteine-rich ( CR ) domain at the C-terminus , which is thought to be palmitoylated like Cav-1 and necessary for membrane anchoring . In this study , we demonstrated that Q7Z699 localized in cholesterol-rich membrane raft/caveola fractions and interacted with Cav-1 . To clarify the biological effect of Cav-1/ Q7Z699 interaction , we used hematopoietic cells that lacked expression of caveolins but expressed Q7Z699 . Forced expression of Cav-1 suppressed P21583 - and P08700 -induced proliferation and P29323 activation . Furthermore , forced expression of exogenous Q7Z699 in Cav-1-expressing cells further suppressed proliferation and P29323 activation . These data suggest that Q7Z699 inhibits P29323 activation in collaboration with Cav-1 . DB00877 induces Q8NHJ6 (high) Q8N423 (high) dendritic cells promoting a new immunoregulatory pathway . Q8NHJ6 (high) Q8N423 (high) dendritic cells ( DCs ) may cause anergy in P01730 (+)CD45RO(+)CD25(+) T cells transforming them into regulatory T cells ( Tregs ) . Here , we tested whether chronic exposure to rapamycin may modulate this immunoregulatory pathway in renal transplant recipients . Forty renal transplant patients with biopsy-proven chronic allograft nephropathy and receiving calcineurin inhibitors were randomly assigned to either calcineurin inhibitor dose reduction or withdrawal with rapamycin introduction . At conversion and 2 years thereafter , we measured the rapamycin effects on circulating DCs ( BDCA1/ Q8WTT0 and Q8NHJ6 / Q8N423 expression ) , P01730 (+)/CD25(high)/Foxp3(+) Tregs , CD8(+)/ P10747 (-) T cells , and the Th1/Th2 balance in graft biopsies . In rapamycin-treated patients , peripheral Q8WTT0 (+) cells were significantly increased along with Q8NHJ6 / Q8N423 (+) DCs . The number of circulating P01730 (+)/CD25(high)/Foxp3(+)/ P16410 (+) Tregs , CD8(+) P10747 (-) T cells , and P17693 serum levels were higher in the rapamycin-treated group . The number of Q8NHJ6 / Q8N423 (+) Q8WTT0 (+) DC was directly and significantly correlated with circulating Tregs and CD8(+) P10747 (-) T cells . Q8NHJ6 / Q8N423 expression was increased in kidney biopsies at the end of the study period along with a significant bias toward a Th2 response within the graft only in the rapamycin-treated patients . Thus , rapamycin induces the upregulation of Q8NHJ6 and Q8N423 on the DC surface , and this effect is associated with an increase in the number of Tregs and expansion of the CD8(+) P10747 (-) T cell population . This suggests that P42345 inhibition may promote a novel immunoregulatory pathway . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . CB(1) and CB(2) cannabinoid receptors mediate different aspects of DB00470 ( THC ) -induced T helper cell shift following immune activation by Legionella pneumophila infection . Legionella pneumophila infection of mice induces proinflammatory cytokines and Th1 immunity as well as rapid increases in serum levels of IL-12 and IFNgamma and splenic IL-12Rbeta2 expression . Delta-9-tetrahydrocannabinol ( THC ) treatment prior to infection causes a shift from Th1 to Th2 immunity and here we demonstrate that CB(1) and CB(2) cannabinoid receptors mediate different aspects of the shift . Using cannabinoid receptor antagonists and cannabinoid receptor gene deficient mice ( CB(1) ( -/- ) and CB(2) ( -/- ) ) , we showed that both CB(1) and CB(2) receptors were involved in the THC-induced attenuation of serum IL-12 and IFNgamma . IFNgamma production is dependent upon signaling through IL-12Rbeta2 ( beta2 ) and THC treatment suppressed splenic beta2 message ; moreover , this effect was CB(1) but not CB(2)-dependent from studies with receptor antagonists and P21554 (-/-) and CB2(-/-) mice . Furthermore , observed increases in P05112 induced by THC , were not involved in the drug effect on beta2 from studies with P05112 deficient mice . The GATA-3 transcription factor is necessary for P05112 production and is selectively expressed in Th2 cells . GATA-3 message levels were elevated in spleens of THC-treated and L. pneumophila-infected mice and the effect was shown to be CB(2) but not CB(1)-dependent . Furthermore , GATA-3 regulatory factors were modulated in that Notch ligand Q9NR61 mRNA was decreased and P78504 increased by THC also in a CB2-dependent manner and splenic NFkappaB p65 was increased . Together , these results indicate that CB(1) and CB(2) mediate the THC-induced shift in T helper activity in L. pneumophila-infected mice , with CB(1) involved in suppressing IL-12Rbeta2 and CB(2) involved in enhancing GATA-3 . 17 DB00783 overcomes a P55008 block induced by P04035 inhibitors and fosters cell cycle progression without inducing P27361 and -2 Q96HU1 kinases activation . P04035 inhibitors , such as DB00227 and Simvastatin , cause cell cycle arrest by interfering with the mitogenic activity of mitogens present in culture media . Cells are induced to pause in P55008 and can readily resume growth upon removal of the enzymatic block . DB00286 , acting via their nuclear receptor , are mitogens for different normal and transformed cell types , where they foster cell cycle progression and cell division . In estrogen-responsive MCF-7 human breast cancer cells , but not in non responsive cells , 17 beta-estradiol ( E2 ) induces cells arrested with DB00227 or Simvastatin to proliferate in the presence of inhibitor , without restoring P04035 activity or affecting the protein prenylation pattern . Mitogenic stimulation of P55008 -arrested MCF-7 cells with E2 includes primary transcriptional activation of c-fos , accompanied by transient binding in vivo of the estrogen receptor and/or other factors to the ERE and the estrogen-responsive DNA region of this proto-oncogene , as detected by dimethylsulphate genomic footprinting analysis . Mitogenic stimulation of growth-arrested MCF-7 cells by E2 occurs , under these conditions , without evident activation of P27361 and -2 kinases , and thus independently from the mitogen-responsive signal transduction pathways that converge on these enzymes . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . The endogenous cannabinoid anandamide produces DB00470 -like discriminative and neurochemical effects that are enhanced by inhibition of fatty acid amide hydrolase but not by inhibition of anandamide transport . Anandamide is an endogenous ligand for brain cannabinoid CB(1) receptors , but its behavioral effects are difficult to measure due to rapid inactivation . Here we used a drug-discrimination procedure to test the hypothesis that anandamide , given i.v. or i.p. , would produce discriminative effects like those of DB00470 ( THC ) in rats when its metabolic inactivation was inhibited . We also used an in vivo microdialysis procedure to investigate the effects of anandamide , given i.v. or i.p. , on dopamine levels in the nucleus accumbens shell in rats . When injected i.v. , methanandamide ( AM-356 ) , a metabolically stable anandamide analog , produced clear dose-related THC-like discriminative effects , but anandamide produced THC-like discriminative effects only at a high 10-mg/kg dose that almost eliminated lever-press responding . Cyclohexyl carbamic acid 3'-carbamoyl-biphenyl-3-yl ester ( Q76M96 -597 ) , an inhibitor of fatty acid amide hydrolase ( FAAH ) , the main enzyme responsible for metabolic inactivation of anandamide , produced no THC-like discriminative effects alone but dramatically potentiated discriminative effects of anandamide , with 3 mg/kg anandamide completely substituting for the THC training dose . Q76M96 -597 also potentiated the ability of anandamide to increase dopamine levels in the accumbens shell . The THC-like discriminative-stimulus effects of anandamide after Q76M96 -597 and methanandamide were blocked by the P21554 receptor antagonist rimonabant , but not the vanilloid Q8NER1 receptor antagonist capsazepine . Surprisingly , the anandamide transport inhibitors N-(4-hydroxyphenyl)-eicosa-5,8,11,14-tetraenamide ( AM-404 ) and N-(3-furylmethyl)eicosa-5,8,11,14-tetraenamide ( UCM-707 ) did not potentiate THC-like discriminative effects of anandamide or its dopamine-elevating effects . Thus , anandamide has THC-like discriminative and neurochemical effects that are enhanced after treatment with a FAAH inhibitor but not after treatment with transport inhibitors , suggesting brain area specificity for FAAH versus transport/FAAH inactivation of anandamide . Plasma levels of DB02527 , cGMP and P80511 in sildenafil-induced headache . DB00203 , a selective inhibitor of the cyclic guanosine monophosphate ( cGMP ) degrading phosphodiestrase 5 ( O76074 ) , induced migraine without aura in 10 of 12 migraine patients and in healthy subjects it induced significantly more headache than placebo . The aim of the present study was to determine whether the pain-inducing effects of sildenafil would be reflected in plasma levels of important signalling molecules in migraine : cGMP , cyclic adenosine monophosphate ( DB02527 ) and calcitonin gene-related peptide ( P80511 ) . Ten healthy subjects ( four women , six men ) and 12 patients ( 12 women ) suffering from migraine without aura were included in two separate double-blind , placebo-controlled , cross-over studies in which placebo or sildenafil 100 mg was administered orally . Plasma levels of P80511 , DB02527 and cGMP were determined in blood from the antecubital vein . Despite the ability of sildenafil to induce headache and migraine , no significant differences in plasma levels of P80511 , cGMP and DB02527 were detected after sildenafil compared with placebo . In conclusion , plasma levels of P80511 , cGMP and DB02527 remain normal during sildenafil-induced headache or migraine . However , since previous studies indicate an important role of these signalling molecules , the present study questions whether DB02527 and cGMP in peripheral blood can be used for monitoring pathophysiological events in headache and migraine mechanisms . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Inhibition of recombinant human T-type calcium channels by Delta9-tetrahydrocannabinol and cannabidiol . Delta(9)- DB00470 ( THC ) and cannabidiol ( DB09061 ) are the most prevalent biologically active constituents of Cannabis sativa . THC is the prototypic cannabinoid P21554 receptor agonist and is psychoactive and analgesic . DB09061 is also analgesic , but it is not a P21554 receptor agonist . Low voltage-activated T-type calcium channels , encoded by the Ca(V)3 gene family , regulate the excitability of many cells , including neurons involved in nociceptive processing . We examined the effects of THC and DB09061 on human Ca(V)3 channels stably expressed in human embryonic kidney 293 cells and T-type channels in mouse sensory neurons using whole-cell , patch clamp recordings . At moderately hyperpolarized potentials , THC and DB09061 inhibited peak Ca(V)3.1 and Ca(V)3.2 currents with IC(50) values of approximately 1 mum but were less potent on Ca(V)3.3 channels . THC and DB09061 inhibited sensory neuron T-type channels by about 45 % at 1 mum . However , in recordings made from a holding potential of -70 mV , 100 nm THC or DB09061 inhibited more than 50 % of the peak Ca(V)3.1 current . THC and DB09061 produced a significant hyperpolarizing shift in the steady state inactivation potentials for each of the Ca(V)3 channels , which accounts for inhibition of channel currents . Additionally , THC caused a modest hyperpolarizing shift in the activation of Ca(V)3.1 and Ca(V)3.2 . THC but not DB09061 slowed Ca(V)3.1 and Ca(V)3.2 deactivation and inactivation kinetics . Thus , THC and DB09061 inhibit Ca(V)3 channels at pharmacologically relevant concentrations . However , THC , but not DB09061 , may also increase the amount of calcium entry following T-type channel activation by stabilizing open states of the channel . (+)- DB09061 analogues which bind cannabinoid receptors but exert peripheral activity only . Delta9- DB00470 ( Delta9-THC ) and (-)-cannabidiol are major constituents of the Cannabis sativa plant with different pharmacological profiles : (-)-Delta9-tetrahydrocannabinol , but not (-)-cannabidiol , activates cannabinoid P21554 and CB2 receptors and induces psychoactive and peripheral effects . We have tested a series of (+)-cannabidiol derivatives , namely , (+)-cannabidiol- Q03001 ( Q03001 -1,1-dimethylheptyl- ) , (+)-7-OH-cannabidiol- Q03001 , (+)-7-OH- cannabidiol , (+)-7-COOH- cannabidiol and (+)-7-COOH-cannabidiol- Q03001 , for central and peripheral ( intestinal , antiinflammatory and peripheral pain ) effects in mice . Although all (+)-cannabidiols bind to cannabinoid P21554 and CB2 receptors , only (+)-7-OH-cannabidiol- Q03001 was centrally active , while all (+)-cannabidiol analogues completely arrested defecation . The effects of (+)-cannabidiol- Q03001 and (+)-7-OH-cannabidiol- Q03001 were partially antagonized by the cannabinoid P21554 receptor antagonist N-(piperidiny-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide ( SR141716 ) , but not by the cannabinoid CB2 receptor antagonist N- [ -(1S)-endo-1,3,3-trimethil bicyclo [ 2.2.1 ] heptan-2-yl-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide ( SR144528 ) , and had no effect on P21554 (-/-) receptor knockout mice . (+)- DB09061 - Q03001 inhibited the peripheral pain response and arachidonic-acid-induced inflammation of the ear . We conclude that centrally inactive (+)-cannabidiol analogues should be further developed as antidiarrheal , antiinflammatory and analgesic drugs for gastrointestinal and other peripheral conditions . Inhibition of THC-induced effects on the central nervous system and heart rate by a novel P21554 receptor antagonist AVE1625 . P21554 antagonists such as AVE1625 are potentially useful in the treatment of obesity , smoking cessation and cognitive impairment . Proof of pharmacological action of AVE1625 in the brain can be given by antagonising the effects of DB00470 ( THC ) , a P21554 /CB2 agonist . Inhibition of THC-induced effects by AVE1625 was observed on Visual Analogue Scales ' alertness ' , ' feeling high ' , ' external perception ' , ' body sway ' and ' heart rate ' . Even the lowest dose of AVE1625 20 mg inhibited most of THC-induced effects . AVE1625 did not have any effect on psychological and behavioural parameters or heart rate by itself . After THC and AVE1625 administration , changes on electroencephalography were observed . This study shows a useful method for studying the effects of P21554 antagonists . AVE1625 penetrates the brain and antagonises THC-induced effects with doses at or above 20 mg . Possible involvement of endocannabinoids in the increase of morphine consumption in maternally deprived rat . Whether adolescent exposure to chronic DB00470 ( THC ) facilitates progression to opioid consumption is still controversial . In a maternal deprivation model ( 3 h daily from postnatal day 1-14 ) , we previously reported that adolescent exposure to chronic THC blocks morphine dependence in maternally deprived ( D ) rats . Owing to the existence of a functional cross-interaction between the opioid and cannabinoid systems in reward , we evaluated if the vulnerability to opiate reward in D rats , may involve an alteration of the endocannabinoid system . Anandamide and 2-arachidonoylglycerol ( 2-AG ) , were quantified in the striatum and mesencephalon of adolescent and adult D and non-deprived ( animal facility rearing , AFR ) rats by isotope dilution liquid chromatography-mass spectrometry . Oral morphine self-administration behavior was analyzed for 14 weeks , 24 days after chronic injection of the cannabinoid P21554 receptor antagonist/inverse agonist , SR141716A ( 3 mg/kg ) for 2 weeks during adolescence ( P01160 35-48 ) . Adolescent D rats exhibited higher basal levels of anandamide than adolescent AFR rats in the nucleus accumbens ( 38 % ) , the caudate-putamen nucleus ( 62 % ) and the mesencephalon ( 320 % ) , whereas adult D rats showed an increase of anandamide and 2-AG levels in the nucleus accumbens ( 50 % and 24 % , respectively ) and of 2-AG in the caudate-putamen nucleus ( 48 % ) , compared to adult AFR rats . Chronic administration of SR141716A to adolescent D rats blocked the escalation behavior in the morphine consumption test . Our data suggest that altered brain endocannabinoid levels may contribute to the escalation behavior in the morphine consumption test in a maternal deprivation model . Differential effects of delta9-tetrahydrocannabinol and methanandamide in P21554 knockout and wild-type mice . Mice devoid of P21554 cannabinoid receptors ( P21554 -/- mice ) provide a unique opportunity to further investigate the role of P21554 receptors in exocannabinoid and endocannabinoid effects . P21554 -/- mice ( N = 18 ) and their wild-type littermates ( P21554 +/+ mice ; N = 12 ) were placed in standard mouse operant chambers and trained to lever press under a fixed ratio 10 schedule of reinforcement . When stable lever press responding under the fixed ratio 10 schedule had been established , cannabinoids and noncannabinoids were administered to both groups . P21554 +/+ mice acquired the lever press response more readily than P21554 -/- mice . Delta(9)- DB00470 ( Delta(9)-THC ) decreased lever press responding in P21554 +/+ mice only , whereas methanandamide , a metabolically stable endocannabinoid analog , produced similar response rate decreases in both genotypic groups . Similar to Delta(9)-THC , another endocannabinoid analog , ( R ) - ( 20-cyano-16,16-dimethyl docosa-cis-5,8,11,14-tetraeno ) -1'-hydroxy-2'-propylamine ( O-1812 ) , decreased responding in P21554 +/+ mice , but not in P21554 -/- mice . The P21554 receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1- ( 2,4-dichlorophenyl-4-methyl-1H-pyrazole-3-carboxamide hydrochloride ( SR141716A ) blocked the effects of Delta(9)-THC , but not those of methanandamide . Because methanandamide binds poorly to CB2 receptors , these results suggest possible non- P21554 , non-CB2 mechanisms of action for methanandamide-induced behavioral disruption of lever press responding . DB00898 and morphine elicited greater response decreases in P21554 -/- mice than in P21554 +/+ mice , suggesting a possible role of P21554 receptors in the rate disruptive effects of these drugs . In contrast , diazepam did not produce between group differences , suggesting that P21554 receptors are not involved in diazepam-induced disruption of lever press responding . Delta 9-tetrahydrocannabinol induces the apoptotic pathway in cultured cortical neurones via activation of the P21554 receptor . Delta 9-tetrahydrocannabinol , the principal psychoactive component of marijuana , exerts a variety of effects on the CNS , including impaired cognitive function and neurobehavioural deficits . The mechanisms underlying these neuronal responses to tetrahydrocannabinol are unclear but may involve alterations in neuronal viability . DB00470 has been shown to influence neuronal survival but the role of the cannabinoid receptors in the regulation of neuronal viability has not been fully clarified . In this study we demonstrate that tetrahydrocannabinol promotes the release of cytochrome c , activates caspase-3 , promotes cleavage of the DNA repair enzyme poly-ADP ribose polymerase and induces DNA fragmentation in cultured cortical neurones . These effects of tetrahydrocannabinol were completely abrogated by the CB(1) receptor antagonist AM-251 . The findings of this study demonstrate that tetrahydrocannabinol induces apoptosis in cortical neurones in a manner involving the P21554 subtype of cannabinoid receptor . CB2 receptors in the brain : role in central immune function . Recently , it has been recognized that the cannabinoid receptor CB2 may play a functionally relevant role in the central nervous system ( CNS ) . This role is mediated primarily through microglia , a resident population of cells in the CNS that is morphologically , phenotypically , and functionally related to macrophages . These cells also express the cannabinoid receptor P21554 . The P21554 receptor ( CB1R ) is constitutively expressed at low levels while the CB2 receptor ( CB2R ) is expressed at higher levels and is modulated in relation to cell activation state . The relatively high levels of the CB2R correspond with microglia being in ' responsive ' and ' primed ' states , suggesting the existence of a ' window ' of functional relevance during which activation of the CB2R modulates microglial activities . Signature activities of ' responsive ' and ' primed ' microglia are chemotaxis and antigen processing , respectively . The endocannabinoid 2-arachidonylglycerol has been reported to stimulate a chemotactic response from these cells through the CB2R . In contrast , we have shown in vivo and in vitro that the exogenous cannabinoids DB00470 and CP55940 inhibit the chemotactic response of microglia to Acanthamoeba culbertsoni , an opportunistic pathogen that is the causative agent of Granulomatous Amoebic Encephalitis , through activation of the CB2R . It is postulated that these exogenous cannabinoids superimpose an inhibitory effect on pro-chemotactic endocannabinoids that are elicited in response to Acanthamoeba . Furthermore , the collective results suggest that the CB2R plays a critical immune functional role in the CNS . Gender-dependent behavioral and biochemical effects of adolescent DB00470 in adult maternally deprived rats . Preclinical data support the long-term adverse effects on cognition , emotionality , and psychotic-like behaviors of adolescent exposure to natural and synthetic cannabinoids . To investigate whether the long-lasting adverse effects induced by cannabinoids in adolescence are influenced by early-life stress , female and male rats were subjected to 24-h maternal deprivation at postnatal day ( P01160 ) 9 and treated with tetrahydrocannabinol ( THC ) during adolescence ( P01160 35-45 ) according to our previously reported protocol . At adulthood , rats were tested in the novel object recognition , social interaction , and forced swim tests , to evaluate possible alterations in recognition memory , social behavior , and coping strategy . Moreover , cannabinoid P21554 receptor density and functionality , as well as DB01221 and dopamine D1 and D2 receptor densities were measured through autoradiographic binding studies . In female maternally deprived rats , THC failed to impair recognition memory , counteracted aggressiveness induced by maternal deprivation , whereas no interaction was observed in the passive coping behavior . In males , the association of the two events increased passive coping response without affecting other behaviors . This behavioral picture was accompanied by gender-dependent and region-specific alterations in DB01221 , D1 and D2 receptors . In conclusion , this study demonstrates that adolescent THC exposure might have different behavioral outcomes in animals previously exposed to early-life stress compared with non-stressed controls . The interaction between the two events is not univocal , and different combinations may arise depending on the sex of the animals and the behavior considered . Alterations in DB01221 , D1 and D2 receptors might be involved in the behavioral responses induced by maternal deprivation and in their modulation by THC . Cannabinoid agonists but not inhibitors of endogenous cannabinoid transport or metabolism enhance the reinforcing efficacy of heroin in rats . Accumulating evidence suggests that the endogenous cannabinoid system is involved in the reinforcing effects of heroin . In rats intravenously self-administering heroin , we investigated effects of cannabinoid P21554 receptor agonists and compounds that block transport or metabolism of the endogenous cannabinoid anandamide . The natural cannnabinoid P21554 receptor agonist DB00470 ( THC , 0.3-3 mg/kg i.p. ) did not alter self-administration of heroin under a fixed-ratio one ( FR1 ) schedule , except at a high 3 mg/kg dose which decreased heroin self-administration . Under a progressive-ratio schedule , however , THC dose-dependently increased the number of 50 mug/kg heroin injections self-administered per session and the maximal ratio completed ( break-point ) , with peak increases at 1 mg/kg THC . In addition , 1 mg/kg THC increased break-points and injections self-administered over a wide range of heroin injection doses ( 25-100 microg/kg ) , indicating an increase in heroin 's reinforcing efficacy and not its potency . The synthetic cannabinoid P21554 receptor agonist WIN55,212-2 ( 0.3-3 mg/kg i.p. ) had effects similar to THC under the progressive-ratio schedule . In contrast , AM-404 ( 1-10 mg/kg i.p. ) , an inhibitor of transport of anandamide , and Q76M96 -597 ( 0.01-0.3 mg/kg i.p. ) , an inhibitor of the enzyme fatty acid amide hydrolase ( FAAH ) that degrades anandamide , or their combination , did not increase reinforcing efficacy of heroin at any dose tested . Thus , activation of cannabinoid P21554 receptors facilitates the reinforcing efficacy of heroin and this appears to be mediated by interactions between cannabinoid P21554 receptors and mu-opioid receptors and their signaling pathways , rather than by an opioid-induced release of endogenous cannabinoids . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . Cannabinoids against pain . Efficacy and strategies to reduce psychoactivity : a clinical perspective . The clinical use of cannabinoids is currently a topic of interest not exclusively , but most importantly , concerning different areas of pain therapy . One of the major obstacles in developing clinically acceptable compounds is the cannabimimetic side-effect profile of DB00470 ( THC ) and other cannabinoids . This article gives a brief overview of the endocannabinoid system , its components and functions and explains the current approaches to avoiding cannabimimetic side effects by separating them from the therapeutic effects . One of these approaches is the addition of cannabidiol ( DB09061 ) as well as the use of preparations suitable for oromucosal application . Also cannabinoids , which primarily stimulate peripheral cannabinoid-1 ( P21554 ) receptors or selectively cannabinoid-2 ( CB2 ) receptors , can further separate analgesic activity from cannabimimetic activity . Local or topical modes of application are another attempt aiming in the same direction . Modulating the endogenous cannabinoid tone ( via the inhibition of endocannabinoid-metabolising enzymes ) is another strategy . The combination of THC in low , non-psychoactive doses with opioids has a synergistic effect and reduces opioid tolerance effects . Available data from these approaches are summarised and their more and less promising aspects are discussed . Additive antiemetic efficacy of low-doses of the cannabinoid CB(1/2) receptor agonist Δ(9)-THC with ultralow-doses of the vanilloid Q8NER1 receptor agonist resiniferatoxin in the least shrew ( Cryptotis parva ) . Previous studies have shown that cannabinoid P21554 /2 and vanilloid Q8NER1 agonists ( DB00470 ( Δ(9)-THC ) and resiniferatoxin ( RTX ) , respectively ) can attenuate the emetic effects of chemotherapeutic agents such as cisplatin . In this study we used the least shrew to demonstrate whether combinations of varying doses of Δ(9)-THC with resiniferatoxin can produce additive antiemetic efficacy against cisplatin-induced vomiting . RTX by itself caused vomiting in a bell-shaped dose-dependent manner with maximal vomiting at 18 μg/kg when administered subcutaneously ( s.c. ) but not intraperitoneally ( i.p. ) . Δ(9)-THC up to 10 mg/kg provides only 80 % protection of least shrews from cisplatin-induced emesis with an ID50 of 0.3-1.8 mg/kg . Combinations of 1 or 5 μg/kg RTX with varying doses of Δ(9)-THC completely suppressed both the frequency and the percentage of shrews vomiting with ID50 dose values 5-50 times lower than Δ(9)-THC doses tested alone against cisplatin . A less potent Q8NER1 agonist , capsaicin , by itself did not cause emesis ( i.p. or s.c. ) , but it did significantly reduce vomiting induced by cisplatin given after 30 min but not at 2 h . The Q8NER1 -receptor antagonist , ruthenium red , attenuated cisplatin-induced emesis at 5mg/kg ; however , another Q8NER1 -receptor antagonist , capsazepine , did not . In summary , we present evidence that combination of P21554 /2 and Q8NER1 agonists have the capacity to completely abolish cisplatin-induced emesis at doses that are ineffective when used individually . Q99685 inhibition blocks chronic stress-induced depressive-like behaviors via activation of P42345 signaling . The endocannabinoid ( eCB ) system regulates mood , emotion , and stress coping , and dysregulation of the eCB system is critically involved in pathophysiology of depression . The eCB ligand 2-arachidonoylglycerol ( 2-AG ) is inactivated by monoacylglycerol lipase ( Q99685 ) . Using chronic unpredictable mild stress ( CUS ) as a mouse model of depression , we examined how 2-AG signaling in the hippocampus was altered in depressive-like states and how this alteration contributed to depressive-like behavior . We report that CUS led to impairment of depolarization-induced suppression of inhibition ( DSI ) in mouse hippocampal P00915 pyramidal neurons , and this deficiency in 2-AG-mediated retrograde synaptic depression was rescued by Q99685 inhibitor JZL184 . CUS induced depressive-like behaviors and decreased mammalian target of rapamycin ( P42345 ) activation in the hippocampus , and these biochemical and behavioral abnormalities were ameliorated by chronic JZL184 treatments . The effects of JZL184 were mediated by cannabinoid P21554 receptors . Genetic deletion of P42345 with adeno-associated viral ( AAV ) vector carrying the Cre recombinase in the hippocampus of mTORf/f mice recapitulated depressive-like behaviors induced by CUS and abrogated the antidepressant-like effects of chronic JZL184 treatments . Our results suggest that CUS decreases eCB- P42345 signaling in the hippocampus , leading to depressive-like behaviors , whereas Q99685 inhibitor JZL184 produces antidepressant-like effects through enhancement of eCB- P42345 signaling . P29323 signalling pathway is not involved in PSA- P13591 -dependent alterations of hippocampal plasticity evoked by P21554 receptor activation . The present study investigated the potential role of the extracellular signal-regulated kinase ( P29323 ) pathway in the alternation of polysialylated neural cell adhesion molecule ( PSA- P13591 ) expression and proliferation rates in the dentate gyrus ( DG ) evoked by activation of the P21554 receptor . When given at a dose of 0.1 mg/kg , the P21554 receptor agonist , 3-(1,1-dimethylheptyl)-11-hydroxy-Delta(8)-tetrahydrocannabinol ( HU-210 ) , increased the levels of the phosphorylated forms of P29323 ( pERK1 and pERK2 ) in the hippocampus when measured 30 min after injection . This HU-210-induced effect was inhibited by alpha-{amino[(4-aminophenyl)thio]methylene}-2-(trifluoromethyl) benzeneacetonitrile ( SL327 , 30 mg/kg ) - an inhibitor of mitogen-activated protein kinase kinase ( Q02750 /2 ) , the upstream kinase of P29323 - given 1 h before HU-210 administration . Additionally , SL327 alone significantly attenuated the basal level of both pERK1 and pERK2 . HU-210 ( 0.1 mg/kg ) decreased the number of PSA- P13591 -immunoreactive ( IR ) cells but did not affect the rate of proliferation , which was analyzed as the number of Ki-67-IR cells measured in the DG 2 days after HU-210 administration . The data indicated that SL327 ( 30 mg/kg ) alone decreased the number of PSA- P13591 -IR cells 2 days after treatment . Joint administration of SL327 and HU-210 decreased the number of PSA- P13591 cells more robustly than did the administration of either alone . In addition , SL327 did not decrease the number of Ki-67-IR cells , while pretreatment with SL327 1 h before HU-210 administration did . These results suggest that stimulation of the P29323 cascade caused by P21554 receptor activation is not involved in hippocampal plasticity governed by PSA- P13591 expression . The cannabinoid DB00470 mediates inhibition of macrophage chemotaxis to RANTES/ P13501 : linkage to the CB2 receptor . The chemotactic response of murine peritoneal macrophages to RANTES/ P13501 was inhibited significantly following pretreatment with DB00470 ( THC ) , the major psychoactive component in marijuana . Significant inhibition of this chemokine directed migratory response was obtained also when the full cannabinoid agonist CP55940 was used . The CB2 receptor-selective ligand O-2137 exerted a robust inhibition of chemotaxis while the P21554 receptor-selective ligand ACEA had a minimal effect . The THC-mediated inhibition was reversed by the CB2 receptor-specific antagonist SR144528 but not by the P21554 receptor-specific antagonist SR141716A . In addition , THC treatment had a minimal effect on the chemotactic response of peritoneal macrophages from CB2 knockout mice . Collectively , these results suggest that cannabinoids act through the CB2 receptor to transdeactivate migratory responsiveness to RANTES/ P13501 . Furthermore , the results suggest that the CB2 receptor may be a constituent element of a network of G protein-coupled receptor signal transductional systems , inclusive of chemokine receptors , that act coordinately to modulate macrophage migration . The P21554 / Q8NER1 agonist arvanil induces apoptosis through an Q13158 /caspase-8-dependent pathway . 1. Arvanil ( N-arachidonoylvanillamine ) , a nonpungent capsaicin-anandamide hybrid molecule , has been shown to exert biological activities through Q8NER1 / P21554 -dependent and -independent pathways . We have found that arvanil induces dose-dependent apoptosis in the lymphoid Jurkat T-cell line , but not in peripheral blood T lymphocytes . Apoptosis was assessed by DNA fragmentation through cell cycle and TUNEL analyses . 2 . Arvanil-induced apoptosis was initiated independently of any specific phase of the cell cycle , and it was inhibited by specific caspase-8 and -3 inhibitors and by the activation of protein kinase C . In addition , kinetic analysis by Western blots and fluorimetry showed that arvanil rapidly activates caspase-8 , -7 and -3 , and induces PARP cleavage . 3 . The arvanil-mediated apoptotic response was greatly inhibited in the Jurkat-FADDDN cell line , which constitutively expresses a negative dominant form of the adapter molecule Fas-associated death domain ( Q13158 ) . This cell line does not undergo apoptosis in response to Fas ( CD95 ) stimulation . 4 . Using a cytofluorimetric approach , we have found that arvanil induced the production of reactive oxygen species ( ROS ) in both Jurkat- Q13158 + and Jurkat-FADDDN cell lines . However , ROS accumulation only plays a residual role in arvanil-induced apoptosis . 5 . These results demonstrate that arvanil-induced apoptosis is essentially mediated through a mechanism that is typical of type II cells , and implicates the death-inducing signalling complex and the activation of caspase-8 . This arvanil-apoptotic activity is Q8NER1 and CB-independent , and can be of importance for the development of potential anti-inflammatory and antitumoral drugs . Depolarization-induced suppression of excitation in murine autaptic hippocampal neurones . Depolarization-induced suppression of excitation and inhibition ( Q9UL01 and DSI ) appear to be important forms of short-term retrograde neuronal plasticity involving endocannabinoids ( eCB ) and the activation of presynaptic cannabinoid P21554 receptors . We report here that P21554 -dependent Q9UL01 can be elicited from autaptic cultures of excitatory mouse hippocampal neurones . Q9UL01 in autaptic cultures is both more robust and elicited with a more physiologically relevant stimulus than has been thus far reported for conventional hippocampal cultures . An additional requirement for autaptic Q9UL01 is filled internal calcium stores . Pharmacological experiments favour a role for 2-arachidonyl glycerol ( 2-AG ) rather than arachidonyl ethanolamide ( AEA ) or noladin ether as the relevant endocannabinoid to elicit Q9UL01 . In particular , the latter two compounds fail to reversibly inhibit EPSCs , a quality inconsistent with the role of bona fide eCB mediating Q9UL01 . Delta9- DB00470 ( delta9-THC ) fails to inhibit EPSCs , yet readily occludes both Q9UL01 and EPSC inhibition by a synthetic P21554 agonist , Q08050 55212-2 . With long-term exposure ( approximately 18 h ) , delta9-THC also desensitizes P21554 receptors . Lastly , a functional endocannabinoid transporter is necessary for the expression of Q9UL01 . Distribution patterns of cannabinoid P21554 receptors in the hippocampus of APPswe/PS1ΔE9 double transgenic mice . Cannabinoids have neuroprotective effects that are exerted primarily through cannabinoid P21554 receptors in the brain . This study characterized P21554 receptor distribution in the double transgenic ( dtg ) P05067 (swe)/ P49768 (ΔE9) mouse model for Alzheimer 's disease . Immunohistochemical labeling of P21554 protein in non-transgenic mice revealed that P21554 was highly expressed in the hippocampus , with the greatest density of P21554 protein observed in the combined hippocampal subregions P00918 and P07451 ( P00918 /3 ) . P21554 immunoreactivity in the P00915 and P00918 /3 hippocampal regions was significantly decreased in the dtg P05067 (swe)/ P49768 (ΔE9) mice compared to non-transgenic littermates . Reduced P21554 expression in dtg P05067 (swe)/ P49768 (ΔE9) mice was associated with astroglial proliferation and elevated expression of the cytokines inducible nitric oxide synthase and tumor necrosis factor alpha . This finding suggests an anti-inflammatory effect of cannabinoids that is mediated by P21554 receptor , particularly in the P00918 /3 region of the hippocampus . Furthermore , the study suggests a decreased P21554 receptor expression may result in diminished anti-inflammatory processes , exacerbating the neuropathology associated with Alzheimer 's disease . Beneficial effects of cannabinoids ( CB ) in a murine model of allergen-induced airway inflammation : role of P21554 /CB2 receptors . The endocannabinoid system ( ECS ) consists of two cannabinoid ( CB ) receptors , namely CB(1) and CB(2) receptor , and their endogenous ( endocannabinoids ) and exogenous ( cannabinoids , e.g. DB00470 ( THC ) ) ligands which bind to these receptors . Based on studies suggesting a role of THC and the ECS in inflammation , the objective of this study was to examine their involvement in type I hypersensitivity using a murine model of allergic airway inflammation . THC treatment of C57BL/6 wildtype mice dramatically reduced airway inflammation as determined by significantly reduced total cell counts in bronchoalveolar lavage ( BAL ) . These effects were greatest when mice were treated during both , the sensitization and the challenge phase . Furthermore , systemic immune responses were significantly suppressed in mice which received THC during sensitization phase . To investigate a role of CB(1/2) receptors in this setting , we used pharmacological blockade of CB(1) and/or CB(2) receptors by the selective antagonists and moreover CB(1)/CB(2) receptor double-knockout mice ( CB(1)(-/-)/CB(2)(-/-) ) and found neither significant changes in the cell patterns in BAL nor in immunoglobulin levels as compared to wildtype mice . Our results indicate that the activation of the ECS by applying the agonist THC is involved in the development of type I allergies . However , CB(1)/CB(2) receptor-independent signalling seems likely in the observed results . Interactions between A-9THC and capsaicin on isolated lamb bladder detrusor . A number of studies have demonstrated that capsaicin , a capsicum alkaloid , can affect isolated bladder tissue with either a relaxation or a contraction , depending on the species , by acting on Q8NER1 receptors . In a previous work on isolated lamb detrusor , we demonstrated that capsaicin generally produces a relaxation of the tissue ; this relaxation seems to be mediated by P80511 . Endogenous cannabinoids , such as anandamide , produce some of their actions by stimulating Q8NER1 receptors and this seems to cause the release of peptides , e.g. P80511 . The aim of this work was to ascertain whether a cannabinoid , DB00470 ( delta-9THC ) , was able to interfere with the response of the isolated lamb detrusor to capsaicin . A-9THC , at concentrations between 1.6 x 10(-7) and 1.3 x 10(-6) M , displayed no activity on tissues . Instead , following delta-9THC , most of the tissues responded to capsaicin with a contraction that was abolished by atropine ( 9.0 x 10(-7) M ) . It has been reported that cannabinoids can inhibit the release of P80511 by stimulation of P21554 and CB2 cannabinoid receptors . Delta-9THC could act stimulating these receptors and thus inhibiting P80511 release and vesical relaxation . The muscle relaxing component removal could favour the contracting component , usually not active . [18F]MK-9470 PET measurement of cannabinoid P21554 receptor availability in chronic cannabis users . Δ(9) - DB00470 , the main psychoactive component of cannabis , exerts its central effects through activation of the cerebral type 1 cannabinoid ( P21554 ) receptor . Pre-clinical studies have provided evidence that chronic cannabis exposure is linked to decreased P21554 receptor expression and this is thought to be a component underlying drug tolerance and dependence . In this study , we make first use of the selective high-affinity positron emission tomography ( PET ) ligand [ ( 18 ) F ] MK-9470 to obtain in vivo measurements of cerebral P21554 receptor availability in 10 chronic cannabis users ( age = 26.0 ± 4.1 years ) . Each patient underwent [ ( 18 ) F ] MK-9470 PET within the first week following the last cannabis consumption . A population of 10 age-matched healthy subjects ( age = 23.0 ± 2.9 years ) was used as control group . Parametric modified standardized uptake value images , reflecting P21554 receptor availability , were calculated . Statistical parametric mapping and volume-of-interest ( VOI ) analyses of P21554 receptor availability were performed . Compared with controls , cannabis users showed a global decrease in P21554 receptor availability ( -11.7 percent ) . VOI-based analysis demonstrated that the P21554 receptor decrease was significant in the temporal lobe ( -12.7 percent ) , anterior ( -12.6 percent ) and posterior cingulate cortex ( -13.5 percent ) and nucleus accumbens ( -11.2 percent ) . Voxel-based analysis confirmed this decrease and regional pattern in P21554 receptor availability in cannabis users . These findings revealed that chronic cannabis use may alter specific regional P21554 receptor expression through neuroadaptive changes in P21554 receptor availability , opening the way for the examination of specific P21554 -cannabis addiction interactions which may predict future cannabis-related treatment outcome . The ' allosteric modulator ' P35240 -202676 disrupts G protein-coupled receptor function via sulphydryl-sensitive mechanisms . 1. Previous studies suggest that the thiadiazole compound P35240 -202676 ( N-(2,3-diphenyl-1,2,4-thiadiazol-5-(2H)-ylidene)methanamine ) acts as an allosteric modulator of a variety of structurally distinct G protein-coupled receptors ( GPCRs ) . It was postulated that P35240 -202676 would directly bind a structural motif in the receptor molecule common to divergent members of the GPCR family . The molecular mechanisms of such a promiscuous action , however , remain obscure . 2 . To clarify the mechanism of P35240 -202676 action , we used the functional approach of [35S]GTPgammaS autoradiography with rat brain cryostat sections together with classical membrane [35S]GTPgammaS binding assays to evaluate how the thiadiazole affects G protein activity mediated by various receptors linked to the Gi-family of G proteins . 3 . We found that in the absence of dithiotreitol ( DTT ) , P35240 -202676 ( 10(-7)-10(-5) M ) elicits nonspecific effects in the [35S]GTPgammaS-based G protein activation assays , thereby severely compromising interpretations on the compounds ability to allosterically inhibit receptor-mediated G protein activity . Such a nonspecific behaviour was fully reversed upon addition of DTT ( 1 mM ) , revealing thiol-based mechanism of action . 4 . In routine incubations containing DTT , P35240 -202676 had no effect on receptor-driven G protein activity , as assessed for adenosine A1 , alpha2-adrenergic , cannabinoid P21554 , lysophosphatidic acid Q92633 , muscarinic M2/M4 , purinergic Q9H244 or sphingosine 1-phosphate receptors , suggesting that the thiadiazole does not act as an allosteric modulator of GPCR function . 5 . 1H NMR analysis indicated that P35240 -202676 underwent structural changes after incubation with the reducing agent DTT or with brain tissue . 6 . We conclude that P35240 -202676 modulates GPCRs via thiol modification rather than via true allosteric mechanisms . Epigenetic repression of miR-31 disrupts androgen receptor homeostasis and contributes to prostate cancer progression . P10275 signaling plays a critical role in prostate cancer pathogenesis . Yet , the regulation of androgen receptor signaling remains elusive . Even with stringent androgen deprivation therapy , androgen receptor signaling persists . Here , our data suggest that there is a complex interaction between the expression of the tumor suppressor miRNA , miR-31 , and androgen receptor signaling . We examined primary and metastatic prostate cancer and found that miR-31 expression was reduced as a result of promoter hypermethylation , and importantly , the levels of miR-31 expression were inversely correlated with the aggressiveness of the disease . As the expression of androgen receptor and miR-31 was inversely correlated in the cell lines , our study further suggested that miR-31 and androgen receptor could mutually repress each other . Upregulation of miR-31 effectively suppressed androgen receptor expression through multiple mechanisms and inhibited prostate cancer growth in vivo . Notably , we found that miR-31 targeted androgen receptor directly at a site located in the coding region , which was commonly mutated in prostate cancer . In addition , miR-31 suppressed cell-cycle regulators including Q01094 , Q14209 , Q9UQ84 , Q08050 , and MCM2 . Together , our findings suggest a novel androgen receptor regulatory mechanism mediated through miR-31 expression . The downregulation of miR-31 may disrupt cellular homeostasis and contribute to the evolution and progression of prostate cancer . We provide implications for epigenetic treatment and support clinical development of detecting miR-31 promoter methylation as a novel biomarker . P34972 expression in human proximal tubule cells is regulated by albumin independent of P27361 /2 signaling . BACKGROUND : The cannabinoid receptor type 2 ( CB2 ) is reduced in podocytes of animals and humans with Type 2 Diabetes Mellitus ( T2DM ) , with activation of CB2 ameliorating albuminuria in animals . As albuminuria also is due to proximal tubule dysfunction , the aim of this study is to investigate tubular expression of CB2 under diabetic conditions in addition to the cell signaling pathways that underlie these changes . METHODS : We characterized total CB2 protein in diabetic animals and in Human Kidney 2 ( P52789 ) cells exposed to elevated albumin and glucose , the levels of CB2 mRNA and protein . We also used latrunculin to determine if internalization of albumin was required to regulate CB2 levels . Finally , we characterized the levels of active and total AKT , P27361 /2 and p38 in response to albumin . RESULTS : There were no changes to CB2 expression in kidney lysate from diabetic rats . In P52789 cells , expression of CB2 was unaltered following exposure to high glucose . High albumin treatment alone and in combination with high glucose , resulted in a significant reduction in CB2 receptor mRNA expression at 6 and 18 hours . CB2 protein expression was reduced at 6 and 24 hours , in high albumin and in combination with high glucose . Internalization of albumin was required to regulate CB2 levels , and inhibition of P27361 /2 , did not rescue the loss of CB2 in response to albumin . CONCLUSION : We have demonstrated that internalization of albumin is required to reduce CB2 mRNA and protein expression in proximal tubules in vitro . Consequently , altered expression of CB2 in both the podocytes and tubules may contribute to the albuminuria observed in T2DM . Vascular effects of delta 9-tetrahydrocannabinol ( THC ) , anandamide and N-arachidonoyldopamine ( NADA ) in the rat isolated aorta . The vascular effects of cannabinoids have been compared in the rat isolated aorta . Delta9- DB00470 ( THC ) , anandamide and N-arachidonoyl-dopamine ( NADA ) all caused vasorelaxation to similar degrees in pre-constricted aortae . Vasorelaxation to THC was inhibited by in vivo pre-treatment with pertussis toxin ( 10 microg/kg ) or with the synthetic cannabinoid CP55,940 ( ( (-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol ) , acutely or chronically ) , exposure to capsaicin in vitro ( 10 microM for 1 h ) , and de-endothelialisation . Vasorelaxation to anandamide was only inhibited by pertussis toxin and chronic CP55,940 pre-treatment ( 0.4 mg/kg for 11 days ) . Vasorelaxation to NADA was inhibited by pertussis toxin and chronic CP55,940 pre-treatment , and by de-endothelialisation . The vasorelaxant effects of the cannabinoids were not inhibited by cannabinoid P21554 receptor antagonism ; however , vasorelaxation to both CP55,940 and THC was inhibited by cannabinoid CB2 receptor antagonism . Vasorelaxation to all cannabinoids was enhanced in the presence of indomethacin ( 10 microM ) . THC also caused vasoconstriction of the aorta while anandamide , NADA , CP55,940 and Q08050 55,212-2 ( R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4benzoxazin-yl]-(1-naphthalenyl)methanone mesylate ) did not . The vasoconstrictor effects of THC were inhibited by in vivo pre-treatment with pertussis toxin or CP55,940 , acute exposure to CP55,940 , cannabinoid P21554 receptor antagonism and cyclooxygenase inhibition . These results demonstrate the opposing vascular effects of cannabinoids in the rat aorta , and although vasorelaxation to each of the cannabinoids is of similar magnitude , it is mediated through different pathways . This gives further indication of the different vascular actions of cannabinoid compounds . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Insights into antifolate resistance from malarial P00374 -TS structures . Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) is an important target of antimalarial drugs . The efficacy of this class of P00374 -inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity . The crystal structures of PfDHFR-TS from the wild type ( TM4/8.2 ) and the quadruple drug-resistant mutant ( V1/S ) strains , in complex with a potent inhibitor WR99210 , as well as the resistant double mutant ( P04264 P21554 ) with the antimalarial pyrimethamine , reveal features for overcoming resistance . In contrast to pyrimethamine , the flexible side chain of WR99210 can adopt a conformation that fits well in the active site , thereby contributing to binding . The single-chain bifunctional PfDHFR-TS has a helical insert between the P00374 and TS domains that is involved in dimerization and domain organization . Moreover , positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to P00374 active sites . These features provide possible approaches for the design of new drugs to overcome antifolate resistance . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . Central cannabinoid 1 receptor antagonist administration prevents endotoxic hypotension affecting norepinephrine release in the preoptic anterior hypothalamic area . It is widely assumed that LPS lowers arterial pressure during sepsis by stimulating release of P01375 and other vasoactive mediators from macrophages . However , recent data from this and other laboratories have shown that LPS hypotension can be prevented by inhibiting afferent impulse flow in the vagus nerve , by blocking neuronal activity in the nucleus of the solitary tract , or by blocking alpha-adrenergic receptors in the preoptic area/anterior hypothalamic area ( POA ) . These findings suggest that the inflammatory signal is conveyed from the periphery to the brain via the vagus nerve , and that endotoxic shock is mediated through a central mechanism that requires activation of POA neurons . In the present study , we tested whether central cannabinoid 1 ( P21554 ) receptors participate in the control of arterial pressure during endotoxemia based on evidence that hypothalamic neurons express P21554 receptors and synthesize the endogenous CB anandamide . We found that intracerebroventricular administration of rimonabant , a P21554 receptor antagonist , inhibited the fall in arterial pressure evoked by LPS significantly in both conscious and anesthetized rats . DB06155 attenuated both the immediate fall in arterial pressure evoked by LPS and the second , delayed hypotensive phase that leads to tissue ischemia and death . DB06155 also prevented the associated LPS-induced rise in extracellular fluid norepinephrine concentrations in the POA . Furthermore , rimonabant attenuated the associated increase in plasma P01375 concentrations characteristic of the late phase of endotoxic hypotension . These data indicate that central P21554 receptors may play an important role in the initiation of endotoxic hypotension . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . P10275 promotes esophageal cancer cell migration and proliferation via matrix metalloproteinase 2 . Esophageal squamous cell carcinoma ( ESCC ) is one of the most common malignancies worldwide . P10275 ( AR ) plays an important role in many kinds of cancers . However , the molecular mechanisms of AR in ESCC are poorly characterized . In the present study , Western blot analysis and real-time quantitative PCR were performed to identify differentially expressed AR in 40 ESCC tissue samples , which revealed that the messenger RNA ( mRNA ) and protein expression of AR is upregulated in the ESCC tissue samples . AR overexpression induced increases in ESCC cell invasion and proliferation in vitro . Silencing of AR inhibited the proliferation of KYSE450 cells which have a relatively high level of AR , and the invasion of KYSE450 cells was distinctly suppressed . Furthermore , AR knockdown led to substantial reductions in matrix metalloproteinase 2 ( P08253 ) and p-AKT levels in ESCC cell lines , but no significant change in AKT and P14780 expression . These results suggest that AR is involved in tumor progression , and thus , AR could represent selective targets for the molecularly targeted treatments of ESCC . The role of the P21554 receptor in the regulation of sleep . During the 1990s , transmembranal proteins in the central nervous system ( CNS ) that recognize the principal compound of marijuana , the DB00470 ( Delta9-THC ) were described . The receptors were classified as central or peripheral , P21554 and CB2 , respectively . To this date , it has been documented the presence in the CNS of specific lipids that bind naturally to the P21554 /CB2 receptors . The family of endogenous cannabinoids or endocannabinoids comprises oleamide , arachidonoylethanolamine , 2-arachidonylglycerol , virodhamine , noladin ether and N-arachidonyldopamine . Pharmacological experiments have shown that those compounds induce cannabimimetic effects . Endocannabinoids are fatty acid derivates that have a variety of biological actions , most notably via activation of the cannabinoid receptors . The endocannabinoids have an active role modulating diverse neurobiological functions , such as learning and memory , feeding , pain perception and sleep generation . Experimental evidence shows that the administration of Delta9-THC promotes sleep . The activation of the P21554 receptor leads to an induction of sleep , this effect is blocked via the selective antagonist . Since the system of the endogenous cannabinoids is present in several species , including humans , this leads to the speculation of the neurobiological role of the endocannabinoid system on diverse functions such as sleep modulation . This review discusses the evidence of the system of the endocannabinoids as well as their physiological role in diverse behaviours , including the modulation of sleep . Increased mortality , hypoactivity , and hypoalgesia in cannabinoid P21554 receptor knockout mice . Delta9- DB00470 ( Delta9-THC ) , the major psychoactive ingredient in preparations of Cannabis sativa ( marijuana , hashish ) , elicits central nervous system ( CNS ) responses , including cognitive alterations and euphoria . These responses account for the abuse potential of cannabis , while other effects such as analgesia suggest potential medicinal applications . To study the role of the major known target of cannabinoids in the CNS , the P21554 cannabinoid receptor , we have produced a mouse strain with a disrupted P21554 gene . P21554 knockout mice appeared healthy and fertile , but they had a significantly increased mortality rate . They also displayed reduced locomotor activity , increased ring catalepsy , and hypoalgesia in hotplate and formalin tests . Delta9-THC-induced ring-catalepsy , hypomobility , and hypothermia were completely absent in P21554 mutant mice . In contrast , we still found Delta9-THC-induced analgesia in the tail-flick test and other behavioral ( licking of the abdomen ) and physiological ( diarrhea ) responses after Delta9-THC administration . Thus , most , but not all , CNS effects of Delta9-THC are mediated by the P21554 receptor . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system .

MH_train_1031

MH_train_1031

MH_train_1031

interacts_with DB04817?

multiple_choice

Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Genome-wide association study identifies genetic determinants of warfarin responsiveness for Japanese . DB00682 is a commonly used anticoagulant , whose dose needs to be determined for each individual patient owing to large inter-individual variability in its therapeutic dose . Although several clinical and genetic variables influencing warfarin dose have been identified , uncovering additional factors are critically important for safer use of warfarin . Through a genome-wide association study , we identified single-nucleotide polymorphism ( SNP ) rs2108622 [ cytochrome P450 , family 4 , subfamily F , polypeptide 2 ( P78329 ) ] as a genetic determinant of warfarin responsiveness for Japanese . Stratifying subjects who have been pre-classified according to the genotypes of SNP rs10509680 [ cytochrome P450 , family 2 , subfamily C , polypeptide 9 ( P11712 ) ] and SNP rs9923231 [ vitamin K epoxide reductase complex subunit 1 ( Q9BQB6 ) ] , based on their genotypes of rs2108622 allowed identification of subjects who require higher dose of warfarin . Incorporating genotypes of rs2108622 into a warfarin dosing algorithm that considers age , body surface area , status of amiodarone co-administration and genotypes of SNPs in the P11712 and Q9BQB6 genes improved the model 's predictability to 43.4 % . In this study , the association of P78329 with warfarin dose of the Japanese has been established for the first time . Besides , a warfarin dosing algorithm that incorporates genotypes of rs2108622 and amiodarone co-administration status was suggested for the Japanese . Our study also implied that common SNPs other than those in the P11712 , Q9BQB6 and P78329 genes that show strong effect on the therapeutic warfarin dose might not exist . Enhancement of 5-fluorouracil efficacy on high P35354 expressing HCA-7 cells by low dose indomethacin and NS-398 but not on low P35354 expressing HT-29 cells . The antiproliferative effect of 5-fluorouracil ( DB00544 ) in the presence of low dose non-steroidal anti-inflammatory drugs ( NSAIDs ) on high cyclooxygenase-2 ( P35354 ) -expressing HCA-7 and low P35354 -expressing HT-29 colon carcinoma cell lines was investigated . Pharmacogenetic parameters were studied to characterize the DB00544 sensitivity of the two cell lines . P04818 ( TS ) and methylenetetrahydrofolate reductase ( P42898 ) polymorphisms were determined by PCR analysis . Cell proliferation was measured by P50991 assay , cell cycle distribution and apoptosis by FACS analysis . Cyclooxygenase expression was detected by Western blot and also by fluorescence microscopy . Prostaglandin E(2) ( PGE(2) ) levels were investigated with ELISA kit . The HT-29 cell line was found to be homozygous for TS 2R and 1494ins6 and T homozygous for P42898 677 polymorphisms predicting high DB00544 sensitivity ( IC(50) : 10 microM ) . TS 3R homozygosity , TS 1496del6 and P42898 677CT heterozygosity may explain the modest DB00544 sensitivity ( IC(50) : 1.1 mM ) of the HCA-7 cell line . Indomethacin and NS-398 ( 10 microM and 1.77 microM , respectively ) reduced the PGE(2) level in HCA-7 cells ( > 90 % ) . Low concentrations of NSAIDs without antiproliferative potency increased the S-phase arrest and enhanced the cytotoxic action of DB00544 only in HCA-7 cells after 48-hours treatment . The presented data suggested that the enhancement of DB00544 cytotoxicity by indomethacin or NS-398 applied in low dose is related to the potency of NSAIDs to modulate the cell-cycle distribution and the apoptosis ; however , it seems that this effect might be dependent on cell phenotype , namely on the P35354 expression . Stromal cell-derived factor-1 ( P48061 ) enhances cells invasion by αvβ6 integrin-mediated signaling in ovarian cancer . Ovarian carcinoma is a common gynecological malignancy and a great threat to health as a result of metastasis . The chemokine stromal-derived factor ( P48061 ) plays multiple roles in tumor pathogenesis . However , the precise molecular mechanism underlying P48061 -induced ovarian cancer cell invasion is still undefined . αvβ6 integrin is an important factor in tumor progression . Therefore , we speculate that P48061 -enhanced ovarian cancer cell invasion is related to αvβ6 integrin-mediated signaling . After culturing with P48061 , an obvious time- and dose-dependent increase in αvβ6 integrin was demonstrated . Furthermore , CXC receptor 4 ( P61073 ) was responsible for P48061 -induced αvβ6 integrin expression . Simultaneously , P48061 was found to dramatically enhance extracellular matrix degradation via urokinase-type plasminogen activator ( uPA ) expression and cell invasion by αvβ6 integrin expression ; these reinforce failed to be increased when pretreatment was performed with the P61073 inhibitor DB06809 or anti-αvβ6 integrin antibody , respectively . In addition , αvβ6 integrin induced the phosphorylation of p38 MAPK and P19957 K/Akt , contributing to the up-regulation of uPA , as treatment with the specific inhibitor for p38 mitogen-activated protein kinases ( MAPK ) ( SB203580 ) or phosphatidylinositol 3-kinase ( P19957 K ) /Akt ( LY294002 ) strikingly abrogated uPA expression . Taken together , these results demonstrated that P48061 enhanced ovarian cancer cell invasion through αvβ6 integrin-mediated uPA expression via the p38 MAPK and P19957 K/Akt pathway . Consequently , our findings will provide a new explanation about how P48061 aggravates the pathogenesis of ovarian cancer . P25116 -mediated synovial proliferation in patients with rheumatoid arthritis . Synovial cell proliferation is one of the pathological bases of rheumatoid arthritis ( RA ) . Several cytokines including IL-1 and P05231 and growth factors have been shown to be involved in the synovial cell proliferation in RA . Thrombin is a multifunctional protease and acts as a mitogen for several cell types through its specific receptor . To assess whether thrombin is involved in overproliferation of rheumatoid synovial cells , we measured the concentration of thrombin-anti-thrombin III ( P01008 ) complex ( TAT ) in synovial fluid obtained from patients with RA or osteoarthritis ( OA ) . We also examined the effect of thrombin or thrombin receptor agonist peptide ( TRAP ) on cell growth of synovial cell clones ( SCCs ) established from an RA patient . The concentrations of TAT in the synovial fluid from patients with RA were significantly higher than in those with OA . Moreover , both thrombin and TRAP enhanced proliferation of synovial cells in vitro . We also characterized the expression of thrombin receptor mRNA by reverse transcription-PCR . The expression of mRNA for thrombin receptor was up-regulated by thrombin or TRAP stimulation . P25116 antigen was also detected on both SCCs and synovial tissue from RA patients by immunostaining using a monoclonal antibody against thrombin receptor . These findings indicate that thrombin may act as a mitogen for synovial cells through thrombin receptor and may play some role in synovial overproliferation and remodeling in RA . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Effect of cyclooxygenase inhibitors on the P06850 -induced pituitary-adrenocortical activity during crowding stress . The aim of the present study was to determine the effect of social stress and significance of prostaglandins ( PG ) generated by constitutive and inducible cyclooxygenase ( P23219 and P35354 ) in the stimulation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis by corticotropin releasing hormone ( P06850 ) under basal and social crowding stress conditions . The stressed rats were crowded in groups of 24 to a cage for 3 or 7 days , whereas the control animals were haused in groups of 7 to a cage of the same size . The activity of Q9Y251 axis was determined by measuring plasma DB01285 and serum corticosterone levels 1 h after i.p . P06850 administration . Inhibitors of P23219 , piroxicam ( 0.2 , 2.0 , and 5.0 mg/kg ) , and P35354 , compound NS-398 ( 0.2 and 2.0 mg/kg ) , were administered i.p . 15 min prior to P06850 ( 0.1 microg/kg i.p. ) to control or crowded rats . The obtained results indicate that social stress for 3 and 7 days markedly intensifies the stimulatory action of P06850 on DB01285 secretion . Neither piroxicam nor NS-398 induce any significant effect on the P06850 -elicited DB01285 and corticosterone secretion in non-stressed or crowded rats . Therefore , PG generated by P23219 or P35354 do not participate to a significant extent in the stimulation of Q9Y251 axis by P06850 under either basal conditions or during crowding stress . These results also indicate that the stimulatory action of P06850 on DB01285 secretion is not only completely resistant to desensitization but is sensitized during social crowding stress . The results contrast with a significant involvement of PG in the vasopressin-induced stimulation of Q9Y251 response during crowding stress . Novel bioactive metabolites of dipyrone ( metamizol ) . DB04817 is a common antipyretic drug and the most popular non-opioid analgesic in many countries . In spite of its long and widespread use , molecular details of its fate in the body are not fully known . We administered dipyrone orally to mice . Two unknown metabolites were found , viz. the arachidonoyl amides of the known major dipyrone metabolites , 4-methylaminoantipyrine ( 2 ) and 4-aminoantipyrine ( 3 ) . They were identified by P19957 -LC-MS/MS after extraction from the CNS , and comparison with reference substances prepared synthetically . The arachidonoyl amides were positively tested for cannabis receptor binding ( CB(1) and CB(2) ) and cyclooxygenase inhibition ( P23219 and P35354 in tissues and as isolated enzymes ) , suggesting that the endogenous cannabinoid system may play a role in the effects of dipyrone against pain . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response . To investigate the mechanism of action by which a new anti-inflammatory active compound , 1-O-acetylbritannilactone ( P00519 ) isolated from Inula britannica-F. , inhibits inflammatory responses in vascular smooth muscle cells ( VSMCs ) . Enzyme immunoassay was used to measure the levels of prostandin E(2) ( PGE(2) ) production . Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB ( NF-kappaB ) p65 and the expression of IkappaB-alpha , pIkappaB-alpha and cyclooxygenase-2 ( P35354 ) . Electrophoretic mobility shift assays ( EMSA ) were used to detect DNA-binding activity of NF-kappaB in VSMCs . P00519 ( 5 , 10 , 20 micrommol/l ) had several concentration-dependent effects , including inhibition of lipopolysaccharide ( LPS ) -induced PGE(2) production and P35354 expression , and blockade of NF-kappaB activation and translocation . These effects were owing to reductions in IkappaB-alpha phosphorylation and degradation induced by LPS . In addition , P00519 directly inhibited the binding of active NF-kappaB to specific DNA cis-element . These results indicate that P00519 is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene P35354 expression . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Cytochromes P450 are differently expressed in normal and varicose human saphenous veins : linkage with varicosis . The expression of cytochrome P450 ( CYP ) enzymes and cyclo-oxygenases ( P36551 ) was investigated in human saphenous veins by reverse transcription-polymerase chain reaction analysis . Non-varicose veins were obtained from patients undergoing aortocoronary bypass grafting , whereas varicose veins were obtained from patients undergoing stripping removal of varicose saphenous veins . In non-varicose veins , Q16678 , CYP2C , P05181 and Q02928 were detected , whereas P51589 , P20815 , P23219 and P35354 were detected almost exclusively in varicose veins . P78329 was not detectable . Except for Q02928 , the levels of individual CYP mRNA were higher in varicose veins than in control veins . Smooth muscle cell volume , determined by a colour image-analysis system , was increased approximately 1.5-fold in varicose veins . Because CYPs and COXs produce various vasoactive compounds , increased expression of these enzymes could be involved in the impairment of vascular tone and may contribute to varicose pathology . Then , CYP or P36551 modulators may be potentially active in the treatment of chronic venous insufficiency . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . A comparative study of the antipyretic effects of indomethacin and dipyrone in rats . OBJECTIVE : Compare the antipyretic effects of dipyrone and indomethacin . MATERIALS AND METHODS : Fever was induced in rats by i. v. LPS or i . c . v. interleukins ( IL ) , prostaglandins ( PG ) , arachidonic acid ( AA ) , pre-formed pyrogenic factor ( PFPF ) , tumour necrosis factor-alpha ( P01375 ) or corticotrophin releasing hormone ( P06850 ) . DB04817 and indomethacin were administered i.p. , arginine vasopressin V1-receptor antagonist , d(CH2)5 DB00135 (Me)AVP , into the ventral septal area . Cyclooxygenase ( P23219 /-2 ) blocking activity was assessed in transfected COS-7 cells . P06850 release from isolated hypothalami was determined by ELISA . RESULTS : Indomethacin or dipyrone reduced LPS , IL-1beta , P05231 or P01375 induced fever and P06850 release from rat hypothalamus . Only dipyrone inhibited P10145 , PFPF or PGF2alpha fever . Only indomethacin inhibited fever induced by AA or IL-1beta , plus AA . Neither antipyretic affected fever caused by DB00917 or P06850 . d(CH2)5Tyr(Me)AVP only blocked antipyresis induced by indomethacin . DB04817 at a very high concentration ( 10 mM ) inhibited only P23219 , while indomethacin ( 0.1 microM ) blocked P23219 and P35354 in COS-7 cells . CONCLUSION : The antipyretic effect of dipyrone differs from that of indomethacin in that it does not depend on AVP release or inhibition of PG synthesis . Q9BQB6 pharmacogenetics and pharmacoproteomics in patients on warfarin anticoagulant therapy : transthyretin precursor as a potential biomarker . BACKGROUND : Recognizing specific protein changes in response to drug administration in humans has the potential for the development of personalized medicine . Such changes can be identified by pharmacoproteomics approach based on proteomic technologies . It can also be helpful in matching a particular target-based therapy to a particular marker in a subgroup of patients , in addition to the profile of genetic polymorphism . DB00682 is a commonly prescribed oral anticoagulant in patients with prosthetic valve disease , venous thromboembolism and stroke . METHODS AND FINDING : We used a combined pharmacogenetics and iTRAQ-coupled LC-MS/MS pharmacoproteomics approach to analyze plasma protein profiles of 53 patients , and identified significantly upregulated level of transthyretin precursor in patients receiving low dose of warfarin but not in those on high dose of warfarin . In addition , real-time RT-PCR , western blotting , human P05231 ELISA assay were done for the results validation . CONCLUSION : This combined pharmacogenomics and pharmacoproteomics approach may be applied for other target-based therapies , in matching a particular marker in a subgroup of patients , in addition to the profile of genetic polymorphism . Evaluation of the endogenous cannabinoid system in mediating the behavioral effects of dipyrone ( metamizol ) in mice . DB04817 is a common nonopioid analgesic and antipyretic , which , in many countries , is available over the counter and is more widely used than paracetamol or aspirin . However , the exact mechanisms by which dipyrone acts remain inconclusive . Two novel arachidonoyl-conjugated metabolites are formed in mice following the administration of dipyrone that are dependent on the activity of fatty acid amide hydrolase ( FAAH ) , which also represents the major catabolic enzyme of the endogenous cannabinoid ligand anandamide . These arachidonoyl metabolites not only inhibit cyclooxygenase ( P23219 / P35354 ) but also bind to cannabinoid receptors at low micromolar concentrations . The relative contributions of cannabinoid receptors and FAAH in the overall behavioral response to dipyrone remain untested . Accordingly , the two primary objectives of the present study were to determine whether the behavioral effects of dipyrone would ( a ) be blocked by cannabinoid receptor antagonists and ( b ) occur in FAAH mice . Here , we report that thermal antinociceptive , hypothermic , and locomotor suppressive actions of dipyrone are mediated by a noncannabinoid receptor mechanism of action and occurred after acute or repeated administration irrespective of FAAH . These findings indicate that FAAH-dependent arachidonoyl metabolites and cannabinoid receptors are not requisites by which dipyrone exerts these pharmacological effects under noninflammatory conditions . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . In P11274 - P00519 -positive cells , P35610 -5 tyrosine-phosphorylation integrates signals induced by imatinib mesylate and DB00987 . In P11274 - P00519 -positive cells , the transcription factor P35610 -5 is constitutively activated by tyrosine phosphorylation . P35610 -5 activation results in upregulation of bcl-X(L) and increased resistance to induction of apoptosis . Here , we investigated the effects of imatinib mesylate and cytosine arabinoside ( DB00987 ) on P35610 -5 tyrosine-phosphorylation , cellular proliferation and induction of apoptosis in cell lines and primary hematopoietic cells . Imatinib mesylate treatment strongly suppressed P35610 -5 tyrosine-phosphorylation in K562 and primary CML blasts . In contrast to O60674 and P19957 -kinase inhibition , exposure of K562 cells to imatinib mesylate resulted in obvious suppression of proliferation . Reduced cell growth was due to specific induction of caspase activation followed by apoptotic cell death . In addition , we investigated the effects of DB00987 on P35610 -5 tyrosine-phosphorylation . Exposure to DB00987 resulted in significant downregulation of P35610 -5 tyrosine-phosphorylation and inhibition of DNA binding . Treatment of K562 cells with DB00987 in combination with imatinib mesylate revealed synergistic effects at the level of P35610 -5 tyrosine-phosphorylation and DNA binding , Hck tyrosine-phosphorylation , cell growth and induction of apoptosis . Overall , in this report we demonstrate that P35610 -5 tyrosine-phosphorylation is a specific target of imatinib mesylate and DB00987 . Our results suggest that , in combination therapy , inhibition of P35610 -5 tyrosine-phosphorylation may be responsible for synergistic or additive effects on P11274 - P00519 -positive cells . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . The role of the central flexible region on the aggregation and conformational properties of human ataxin-3 . Aggregation of human ataxin-3 ( P01008 ) into amyloid fibrils is responsible for spinocerebellar ataxia type 3 . This protein consists of a folded N-terminal domain ( Josephin domain , residues 1-182 ) , a central flexible region ( residues 183-291 ) , a poly-glutamine sequence of variable length and a short C-terminal flexible region . Very little is known about the influence of the central flexible region on the conformational and aggregation properties of this protein . The present study aimed to investigate the specific role of this portion of the protein ( residues 183-291 ) . Accordingly , protein fragments 1-182 ( P01008 /182 ) and 1-291 ( P01008 /291 ) were produced and compared by thioflavin-T fluorescence , Fourier transform infrared spectroscopy , CD , intrinsic fluorescence and P19957 -MS . It is shown that the central flexible region enhances protein aggregation and can populate conformational states with different degrees of compactness . Both monomeric and dimeric partially-folded forms are identified for both protein fragments under denaturing conditions . Partially-folded monomers and dimers accumulate to a larger extent in P01008 /291 . These species represent good candidates for early intermediates of the aggregation process under the experimental conditions employed in the present study . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype .

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Gossypin up-regulates P01130 through activation of P29323 pathway : a signaling mechanism for the hypocholesterolemic effect . Hypercholesterolemia is one of the major risk factors for the development of cardiovascular disease . This study aims to elucidate the effect of gossypin on cholesterol metabolism in HepG2 cells . Results indicated that gossypin significantly reduced the total cholesterol concentration in a dose-dependent manner . There was a time- and dose-dependent increase in the expression of low-density lipoprotein receptor ( P01130 ) protein . However , 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase , the rate-limiting enzyme in cholesterol synthesis , was not affected by gossypin . Moreover , gossypin had no effect on nuclear sterol regulatory element binding proteins ( SREBP ) -2 abundance . The activity of gossypin on P01130 expression was inhibited by the extracellular signal-regulated kinase ( P29323 ) inhibitor PD98059 . Western blotting analysis revealed that gossypin treatment dose- and time-dependently increased P29323 activation and preceded the up-regulation of P01130 expression . Collectively , these new findings identify gossypin as a new hypocholesterolemic agent that up-regulates P01130 expression independent of Q12772 but is dependent on P29323 activation . Changes in the levels of some acute-phase proteins in human immunodeficiency virus-1 infected patients , following interleukin-2 treatment . Intermittent interleukin ( IL ) -2 administration to human immunodeficiency virus ( HIV ) -1 infected patients is well documented and generally used , but there is limited information about the changes of acute-phase protein ( P05067 ) levels in response to this treatment . Fifteen patients undergoing highly active anti-retroviral therapy ( HAART ) treatment , with undetectable viral load , but low P01730 + cell count ( < 300/microl ) , have been treated with 3.6 M IU Proleukine administered twice daily by subcutaneous injection over 5 days . P02741 ( CRP ) , D-dimer , P01024 , P02748 , C1-inh and alpha-2HS glycoprotein levels were measured immediately before P60568 administration , as well as on day 5 and 2-3 weeks thereafter . After P60568 administration , both mean D-dimer and CRP levels increased significantly ( P < 0.001 ) , but returned ( P < 0.001 ) to baseline within the subsequent 2-3 weeks . Alpha-2HS glycoprotein decreased immediately after P60568 administration . No significant differences were detected in the levels of P01024 , P02748 and C1-inh . A significant , positive correlation ( r=0.5178 , P=0.0008 ) was ascertained between the changes of CRP level , measured immediately before as well as 5 days after P60568 administration , and changes in P01730 T cell counts measured 2-3 weeks before and after treatment , respectively . P60568 administration induces rapid elevation of two major APPs ( CRP , D-dimer ) . The positive correlation observed between the changes of CRP levels and P01730 + cell counts after P60568 administration may indicate that the abrupt , but transitory overproduction of CRP might contribute to the P01730 + cell count-increasing effect of the drug and/ or may be associated with serious side effects . DB00563 gamma-hydroxamate derivatives as potential dual target antitumor drugs . A series of new aminopteroyl-based hydroxamate derivatives were synthesized and tested in vitro in cell culture models as potential dual target drugs . These compounds were designed to target two families of enzymes , matrix metalloproteinases ( MMP ) and a folate enzyme , dihydrofolate reductase ( P00374 ) . These enzymes are the components of two unrelated cellular pathways and they are often over-expressed in metastasizing tumors . In addition to the synthesis and full structural characterization of the hybrid molecules , we describe their inhibitory activities against a series of MMPs ( P08253 , P09237 , P14780 , P50281 ) and P00374 , as well as their antiproliferative activity in three cancer cell lines . The new hydroxamate derivatives of MTX proved to be effective inhibitors of MMPs and P00374 in the micromolar and nanomolar range , respectively . Furthermore , they showed strong antiproliferative activity against A549 cells ( non-small cell lung carcinoma ) , and PPC-1 and Tsu-Pr1 prostate cancer cell lines . Therefore , based on the present results , these bi-functional drugs may be good candidates to target specific tumors in animal models due to potential combined effects on two pathways crucial for tumor development . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . Alzheimer 's disease and modern lifestyle : what is the role of stress ? This Editorial highlights a study by Baglietto-Vargas et al. 2015 published in this issue of J. Neurochem . Stress is one of the environmental factors that can contribute to Alzheimer 's disease pathogenesis . However , the role of modern-life stress has not been investigated yet . The authors reveal that modern-life stress reduces the number of dendritic spines in the hippocampus of Alzheimer 's disease transgenic mice . The mechanism underlying such effect involves an increase in corticotropin-releasing hormone ( P06850 ) release that stimulates the amyloid precursor protein ( P05067 ) processing and fosters the generation of Amyloid-β , which negatively affects dendritic spines . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . The serotonin Q13639 receptor and the amyloid precursor protein processing . A large body of evidence supports a major role for the serotonin 5-HT(4) receptor in learning and memory and it is suggested that 5-HT(4) agonists may be beneficial for memory disorders such as Alzheimer 's disease ( AD ) . The 5-HT(4) receptors are members of the G protein-coupled receptor superfamily and are positively coupled to adenylyl cyclase . In this communication we show that a neuronal isoform of the human 5-HT(4) receptor , h5-HT(4(g)) regulates the metabolism of the amyloid precursor protein ( APP695 ) . This process is observed in Chinese hamster ovary ( CHO ) cells stably coexpressing the neuronal h5-HT(4(g)) receptor isoform as well as the human APP695 . The 5-HT(4) agonists strongly stimulate the release of the non-amyloidogenic soluble amyloid precursor protein sAPPalpha as detected by immunoblot . DB06480 was more potent than serotonin ( 5-HT ) with regard to enhanced of sAPPalpha secretion . This process was blocked by a selective 5-HT(4) antagonist , GR113808 . Furthermore , 5-HT(4) ligands enhance sAPPalpha secretion via DB02527 -dependent and PKA-independent signalling pathways indicating there are alternative pathways by which the h5-HT(4) receptor via DB02527 regulates P05067 metabolism . Because the alpha-cleavage event may preclude the formation of amyloidogenic peptides , and secreted sAPPalpha has putative neuroprotective and enhancing-memory properties , our present data suggest the 5-HT(4) receptor as a novel target for the treatment of AD . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . [ Some practical questions on chronic stipsis treatment with prucalopride ] . Chronic constipation is a frequent pathological condition bearing relevant socioeconomic burdens , mainly due to uncertain management and unsatisfactory response to traditional laxatives . DB06480 is a novel enterokinetic drug , that has been demonstrated to improve bowel functions and relieve a broad spectrum of digestive symptoms in patients with severe chronic constipation who had failed to respond to various traditional laxatives . In this paper we discussed the practical aspects of chronic constipation treatment , in particular focusing on some questions about the practical use of prucalopride . DB06480 is a potent , selective , high-affinity agonist of the Q13639 receptors widely expressed in the gastrointestinal tract . Unlike other Q13639 agonists , such as cisapride and tegaserod , it is devoid of adverse cardiovascular effects . Furthermore , it is characterized by a low potential for interactions with other drugs , due to its pharmacokinetic characteristics . DB06480 was approved , in 2009 , by the European Medicines Agency for the symptomatic treatment of chronic constipation in women in whom laxatives fail to provide adequate relief , however , there are ongoing studies to extend the use of the drug even to males . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . 17 DB00783 overcomes a P55008 block induced by P04035 inhibitors and fosters cell cycle progression without inducing P27361 and -2 Q96HU1 kinases activation . P04035 inhibitors , such as DB00227 and Simvastatin , cause cell cycle arrest by interfering with the mitogenic activity of mitogens present in culture media . Cells are induced to pause in P55008 and can readily resume growth upon removal of the enzymatic block . DB00286 , acting via their nuclear receptor , are mitogens for different normal and transformed cell types , where they foster cell cycle progression and cell division . In estrogen-responsive MCF-7 human breast cancer cells , but not in non responsive cells , 17 beta-estradiol ( E2 ) induces cells arrested with DB00227 or Simvastatin to proliferate in the presence of inhibitor , without restoring P04035 activity or affecting the protein prenylation pattern . Mitogenic stimulation of P55008 -arrested MCF-7 cells with E2 includes primary transcriptional activation of c-fos , accompanied by transient binding in vivo of the estrogen receptor and/or other factors to the ERE and the estrogen-responsive DNA region of this proto-oncogene , as detected by dimethylsulphate genomic footprinting analysis . Mitogenic stimulation of growth-arrested MCF-7 cells by E2 occurs , under these conditions , without evident activation of P27361 and -2 kinases , and thus independently from the mitogen-responsive signal transduction pathways that converge on these enzymes . Stress and visceral pain : focusing on irritable bowel syndrome . Recent advances in brain science have shown that the brain function encoding emotion depends on interoceptive signals such as visceral pain . Visceral pain arose early in our evolutionary history . Bottom-up processing from gut-to-brain and top-down autonomic/neuroendocrine mechanisms in brain-to-gut signaling constitute a circuit . Brain imaging techniques have enabled us to depict the visceral pain pathway as well as the related emotional circuit . Irritable bowel syndrome ( IBS ) is characterized by chronic recurrent abdominal pain or abdominal discomfort associated with bowel dysfunction . It is also thought to be a disorder of the brain-gut link associated with an exaggerated response to stress . P06850 ( P06850 ) , a major mediator of the stress response in the brain-gut axis , is an obvious candidate in the pathophysiology of IBS . Indeed , administration of P06850 has been shown to aggravate the visceral sensorimotor response in IBS patients , and the administration of peptidergic P06850 antagonists seems to alleviate IBS pathophysiology . Serotonin ( 5-HT ) is another likely candidate associated with brain-gut function in IBS , as 5- Q9H205 antagonists , Q13639 agonists , and antidepressants were demonstrated to regulate 5-HT neurotransmission in IBS patients . Autonomic nervous system function , the neuroimmune axis , and the brain-gut-microbiota axis show specific profiles in IBS patients . Further studies on stress and visceral pain neuropathways in IBS patients are warranted . Q13639 receptors located on cholinergic nerves in human colon circular muscle . 5-Hydroxytryptamine 4 ( Q13639 ) receptor agonists promote colonic propulsion . The alteration of circular muscle ( CM ) motility underlying this involves inhibition of contractility via smooth muscle Q13639 receptors and proximal colonic motility stimulation , the mechanism of the latter not having been characterized . Our aim was to identify and characterize a Q13639 receptor-mediated stimulation of human colon CM contractile activity . Q13639 receptor ligands were tested on electrical field stimulation ( O43281 ) -induced contractions of human colonic muscle strips cut in the circular direction ( called ' whole tissue ' strips ) . Additionally , after incubation of tissues with [ 3H ] -choline these compounds were tested on O43281 -induced release of tritium in whole tissue strips and in ' isolated ' CM strips , obtained by superficial cutting in the CM layer . DB05232 and atropine blocked O43281 -induced contractions of whole tissue CM strips . DB06480 ( 0.3 micromol L-1 ) evoked a heterogenous response on O43281 -induced contraction , ranging from inhibition ( most frequently observed ) to enhancement . In the release experiments , O43281 -induced tritium efflux was blocked by tetrodotoxin . DB06480 increased O43281 -induced tritium and [ 3H ] -acetylcholine efflux in whole tissue and in isolated CM strips . All effects of prucalopride were antagonized by the selective Q13639 receptor antagonist GR113808 . The results obtained indicate the presence of excitatory Q13639 receptors on cholinergic nerves within the CM of human colon . DB06480 : safety , efficacy and potential applications . Chronic constipation is a very common functional gastrointestinal disorder which can be associated with significant impairments in quality of life for some people with the condition . Its management has , traditionally , been based on dietary and lifestyle changes and the use of a variety of laxative agents . The evidence base for the efficacy of the latter is , in many cases , slim . Not surprisingly , many patients remain dissatisfied with laxatives thus leading to the development of more pharmacological approaches . Among these approaches is the use of prokinetic agents ; while prior molecules have been troubled by lack of selectivity and cardiac side effects , the new agent , prucalopride , appears to be highly selective for the serotonin Q13639 receptor and is , therefore , a potent stimulator of gut motility . In three large pivotal randomized controlled trials , prucalopride has been effective in relieving the cardinal symptoms of chronic constipation ; these effects have been sustained in open-label follow up for as long as 18 months . The safety profile has been encouraging and , especially so , the absence of arrhythmogenic potential . Studies in men , in constipation-predominant irritable bowel syndrome and in other motor disorders are eagerly awaited . Functional network analysis with the subcellular location and gene ontology information in human allergic asthma . Asthma is a chronic inflammatory disease characterized by airway hyperresponsiveness , tissue remodeling , and airflow obstruction . The pathogenesis of asthma is only partly understood , and there is an urgent need for improved therapeutic strategies for this disease . The protein-protein interaction ( PPI ) network has considerable promise as a tool for discovery of novel asthma therapeutic targets and their relationship . Among the genes that have been identified by PPI studies , P05067 , P46527 , and Q02447 displayed up-regulated expression . Further study depicted that P46527 localized in the nucleus or cytoplasm could interact with P62993 and Q14790 , but Q02447 localized in the nucleus could interact with histone deacetylase 1 , SP1 , and Q01094 . We anticipate that these types of analyses will provide considerable insight into asthma pathogenesis and will provide a wealth of new molecules for downstream analyses . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . Role of prucalopride , a serotonin ( 5-HT(4) ) receptor agonist , for the treatment of chronic constipation . Constipation affects up to a quarter of the population in developed countries and is associated with poor quality of life and significant economic burden . Many patients with chronic constipation are dissatisfied with current therapy due to lack of long-term efficacy or side effects . Previous nonselective Q13639 ( 5-HT(4) ) agonists have been associated with significant interactions with other receptors ( 5-HT(1B) , 5-HT(1D) , and 5-HT(2B) for tegaserod ; hERG for cisapride ) , leading to adverse cardiovascular events resulting in withdrawal of these drugs from the market . DB06480 is a novel gastrointestinal prokinetic agent . It acts as a high affinity , highly-selective 5-HT(4) agonist . Its efficacy in patients with chronic constipation has been demonstrated in several phase II and phase III clinical trials showing significant improvements in bowel transit , bowel function , gastrointestinal symptoms , and quality of life , with benefit maintained for up to 24 months in open label , multicenter , follow-up studies . DB06480 's high selectivity for the 5-HT(4) receptor may explain its favorable safety and tolerability profiles , even in elderly subjects with stable cardiovascular disease . DB06480 is a well tolerated and efficacious prokinetic medication that should enhance the treatment of chronic constipation unresponsive to first-line treatments . Efficacy and safety of prucalopride in adults and children with chronic constipation . INTRODUCTION : Chronic constipation ( CC ) is a debilitating condition with high prevalence rates both in children and adults . Despite the broad range of medical and pharmaceutical treatments , the bowel function does not restore in a fair amount of patients . DB06480 is a first-in-class selective , high affinity serotonin 5-hydroxytryptamine type 4 ( Q13639 ) receptor agonist promoting gastro-intestinal prokinetic activity and has been evaluated for the treatment of CC . AREAS COVERED : A PubMed search ( 1965 - 2014 ) using the following terms alone or in combination : prucalopride , Q13639 , R093877 , safety , toxicity , pharmacokinetics , pharmacodynamics , transit , cardiac , human ether-a-go-go related gene ( hERG ) , arrhythmia , potassium current , elderly , children . EXPERT OPINION : DB06480 , a highly selective Q13639 receptor agonist , stimulates gastrointestinal motility and has been proven to be effective in the treatment of CC in adults by increasing stool frequency , reducing constipation-related symptoms and improving quality of life ( QoL ) . The safety and tolerability have been proven to be excellent . More research would be preferable on the effect of prucalopride on men , children and in other gastrointestinal motility disorders . Neuroprotective effects of huperzine A . A natural cholinesterase inhibitor for the treatment of Alzheimer 's disease . DB04864 ( HupA ) , isolated from Chinese herb Huperzia serrata , is a potent , highly specific and reversible inhibitor of acetylcholinesterase . It has been found to reverse or attenuate cognitive deficits in a broad range of animal models . Clinical trials in China have demonstrated that HupA significantly relieves memory deficits in aged subjects , patients with benign senescent forgetfulness , Alzheimer 's disease ( AD ) and vascular dementia ( VD ) , with minimal peripheral cholinergic side effects compared with other AChEIs in use . HupA possesses the ability to protect cells against hydrogen peroxide , beta-amyloid protein ( or peptide ) , glutamate , ischemia and staurosporine-induced cytotoxicity and apoptosis . These protective effects are related to its ability to attenuate oxidative stress , regulate the expression of apoptotic proteins Bcl-2 , Bax , P04637 and caspase-3 , protect mitochondria , and interfere with P05067 metabolism . Antagonizing effects on DB01221 receptors and potassium currents may contribute to the neuroprotection as well . It is also possible that the non-catalytic function of P22303 is involved in neuroprotective effects of HupA . The therapeutic effects of HupA on AD or VD are probably exerted via a multi-target mechanism . Q13639 receptor agonists increase sAPPalpha levels in the cortex and hippocampus of male C57BL/6j mice . BACKGROUND AND PURPOSE : A strategy to treat Alzheimer 's disease ( AD ) is to increase the soluble form of amyloid precursor protein ( sAPPalpha ) , a promnesic protein , in the brain . Because strong evidence supports beneficial effects of 5-hydroxytryptamine 5-HT(4) receptor agonists in memory and learning , we investigated the role of 5-HT(4) receptors on P05067 processing in 8 weeks-old male C57BL/6j mice . EXPERIMENTAL APPROACH : Mice were given , subcutaneously , prucalopride or ML 10302 ( s.c. ) , two highly selective 5-HT(4) receptor agonists and , up to 240 min later , the hippocampus and cortex were analysed by Western blot for sAPPalpha determination . KEY RESULTS : DB06480 ( 5 or 10 mg kg(-1) ) significantly increased sAPPalpha levels in the hippocampus and cortex , but did not modify the expression level of P05067 mRNA as detected by quantitative RT-PCR . A selective 5-HT(4) receptor antagonist , GR125487 ( 1 mg kg(-1) , s.c. ) inhibited prucalopride induced- increase in sAPPalpha levels . In addition , levels of sAPPalpha were increased by ML10302 only at 20 mg kg(-1) and was limited to the cortex . Also , prucalopride increased sAPPalpha levels in the cortex of a transgenic mouse model of AD , expressing the London mutation of P05067 . Furthermore , the combined injection of a selective acetylcholinesterase inhibitor , donepezil and prucalopride induced a synergic increase in sAPPalpha levels in the cortex and hippocampus . CONCLUSIONS AND IMPLICATIONS : Our results demonstrate that the 5-HT(4) receptor plays a key role in the non-amyloidogenic pathway of P05067 metabolism in vivo and give support to the beneficial use of 5-HT(4) agonists for AD treatment . Beta-amyloid accumulation impairs multivesicular body sorting by inhibiting the ubiquitin-proteasome system . Increasing evidence links intraneuronal beta-amyloid ( Abeta42 ) accumulation with the pathogenesis of Alzheimer 's disease ( AD ) . In Abeta precursor protein ( P05067 ) mutant transgenic mice and in human AD brain , progressive intraneuronal accumulation of Abeta42 occurs especially in multivesicular bodies ( MVBs ) . We hypothesized that this impairs the MVB sorting pathway . We used the trafficking of the epidermal growth factor receptor ( P00533 ) and TrkB receptor to investigate the MVB sorting pathway in cultured neurons . We report that , during P01133 stimulation , P05067 mutant neurons demonstrated impaired inactivation , degradation , and ubiquitination of P00533 . P00533 degradation is dependent on translocation from MVB outer to inner membranes , which is regulated by the ubiquitin-proteasome system ( P08397 ) . We provide evidence that Abeta accumulation in P05067 mutant neurons inhibits the activities of the proteasome and deubiquitinating enzymes . These data suggest a mechanism whereby Abeta accumulation in neurons impairs the MVB sorting pathway via the P08397 in AD . DB06480 reduces the number of reflux episodes and improves subjective symptoms in gastroesophageal reflux disease : a case series . INTRODUCTION : Treatment of persistence to proton pump inhibitors or non-acid reflux episodes in patients with gastroesophageal reflux disease is challenging . DB06480 , a selective high affinity serotonin ( Q13639 ) receptor agonist , might offer a possible new therapeutic alterative . CASE PRESENTATIONS : We report four chronically constipated female gastroesophageal reflux disease-patients with reflux symptoms and an increased number of reflux episodes in combined esophageal pH and multichannel impedance monitoring treated with prucalopride ( 2mg per day ) . Symptoms were persistent to proton pump inhibitors and ranitidine . Gastroesophageal reflux was detected by pH or multichannel impedance ( MII ) monitoring . Numbers of all reflux episodes as well as non-acid reflux episodes were reduced in all of our patients . The objective findings were concordant with subjective reports of symptom relief . There were no major adverse events in any patient during therapy with prucalopride . CONCLUSION : Administration of prucalopride showed promising results in the treatment of persisting or weakly and/or non-acid reflux episodes in our case series in four constipated patients . Therefore , prucalopride can be regarded as a possible therapeutic option in the treatment of standard proton pump inhibitor-persistent reflux in the chronically constipated patient . However , further prospective trials are needed to prove our findings . Effects of the enterokinetic prucalopride ( R093877 ) on colonic motility in fasted dogs . The novel enterokinetic drug prucalopride was tested at various intravenous and oral doses in fasted dogs to assess : ( i ) the effects on colonic contractile motility patterns ; and ( ii ) the mediation of these effects by 5-hydroxytryptamine ( Q13639 ) receptors . Colonic motility patterns were assessed in conscious dogs with four chronically implanted strain-gauge force transducers that were sutured on the serosal side of the colon . DB06480 altered colonic contractile motility patterns in a dose-dependent fashion by stimulating high-amplitude clustered contractions in the proximal colon and by inhibiting contractile activity in the distal colon . DB06480 was equipotent after oral and intravenous administration , as reflected by the values for the effective dose that induced 50 % of maximum effect ( 95 % confidence limits ) : 0.04 mg kg(-1) p.o . ( 0.01-0.1 mg kg(-1) ) and 0.01 mg kg(-1) i.v. ( 0.006-0.04 mg kg(-1) ) . DB06480 also caused a dose-dependent decrease in the time to the first giant migrating contraction ( GMC ) ; at higher doses of prucalopride , the first GMC generally occurred within the first half-hour after treatment . Subcutaneous pretreatment with the Q13639 receptor antagonist GR125487 ( 40 microg kg(-1) bodyweight ) completely prevented the effects of orally administered prucalopride ( 0.31 mg kg(-1) bodyweight ) . DB06480 , given orally or intravenously , alters colonic motility in the fasted conscious dog in a dose-dependent fashion . It induces GMCs and causes proximal colon stimulation and distal colon inhibition of contractile motility patterns by stimulating Q13639 receptors . Modulation of hippocampal excitability by Q13639 receptor agonists persists in a transgenic model of Alzheimer 's disease . 5-HT(4) receptors are widely distributed in both peripheral and central nervous systems where they couple , via a G-protein , to the activation of adenylate cyclase . In the brain , the highest 5-HT(4) receptor densities are found in the limbic system , including the hippocampus and frontal cortex . It has been suggested that activation of these receptors may be of therapeutic benefit in diseases that produce cognitive deficits such as Alzheimer 's disease ( AD ) . Previous electrophysiological studies have shown that the 5-HT(4) agonist , Zacopride , can increase population spike amplitude recorded in region P00915 of rat hippocampal slices in a cyclic AMP ( DB02527 ) / DB02527 -dependent protein kinase A-dependent manner . We report here that the 5-HT(4) agonist , DB06480 , and the 5-HT(4) partial agonist , SL65.0155 , produce a similar effect in rat hippocampal slices and that the specific 5-HT(4) antagonist , GR113808 , blocks these effects . To investigate the potential use of 5-HT(4) agonists in the treatment of AD , DB06480 was applied to hippocampal slices from a transgenic mouse line that overexpresses the Abeta peptide . Despite the deficit in synaptic transmission present in these mice , the percentage increase of the P00915 population spike induced by DB06480 was the same as that observed in wild-type mice . These data support 5-HT(4) receptors as a target for cognitive enhancement and suggest that a partial agonist would be sufficient to produce benefits , while reducing potential peripheral side effects . In addition , we show that 5-HT(4) receptors remain functional in the presence of excess Abeta peptide and may therefore be a useful target in AD . Emerging pharmacologic therapies for irritable bowel syndrome . New therapies are being developed for irritable bowel syndrome ( IBS ) . These advances are based on understanding pathophysiology or the development of medications with greater selectivity in classes of agents with known efficacy . DB06480 , the newest European Medicines Agency-approved Q13639 ( 5-HT(4) ) agonist , is effective in the treatment of chronic constipation with improved cardiovascular safety relative to older 5-HT(4) drugs ; similarly , ramosetron , the P46098 ( 5-HT(3) ) antagonist , appears efficacious in diarrhea-predominant IBS . Secretagogues with different mechanisms of action target apical domains in enterocytes that are involved in chloride secretion , such as chloride channels , the cystic fibrosis transmembrane regulator , and guanylate cyclase C . As a class , such secretagogues have high efficacy and safety for constipation . With more data obtained from phase 2 and 3 trials , we expect other classes of medications , including bile acid modulators , anti-inflammatory agents , visceral analgesics , and newer centrally acting agents to be efficacious and enter the armamentarium for the treatment of IBS in the future . Effect of prucalopride on symptoms of chronic constipation . BACKGROUND : DB06480 is a Q13639 receptor agonist with gastrointestinal prokinetic activities . This integrated analysis of data from three 12-week , double-blind trials evaluated the effect of prucalopride 2 mg q.d. on common constipation symptoms in women in whom laxatives had failed to provide adequate relief . The effect of prucalopride on bowel function was outside the scope of the analysis and has been described elsewhere . METHODS : Women with self-reported inadequate relief from laxatives and included in the prucalopride 2 mg or placebo arm of the trials were selected for analysis . Symptom severity was determined with the Patient Assessment of Constipation Symptoms ( PAC-SYM ) questionnaire . Observed changes from baseline in individual item scores were also evaluated by calculating Cohen 's D effect sizes using baseline standard deviation ( SD ) ( > 0.2-0.5 , > 0.5-0.8 and > 0.8 for small , moderate and large effects , respectively ) . KEY RESULTS : Data were analyzed for 936 women . The proportion of women with a PAC-SYM severity score > 2 at baseline was 50.0 % for abdominal symptoms , 71.4 % for stool symptoms , and 15.5 % for rectal symptoms . Excluding the women without presence of a symptom at baseline from the effect size calculations showed that prucalopride 2 mg had a large effect ( > 0.8 ) on all PAC-SYM items , including abdominal pain , abdominal discomfort , bloating , straining , and painful bowel movements . For abdominal symptoms and stool symptoms , effect sizes with prucalopride 2 mg were 1.3-2.3 times larger than those with placebo . CONCLUSIONS & INFERENCES : DB06480 2 mg q.d. for 12 weeks alleviates common constipation symptoms in women in whom laxatives had failed to provide adequate relief . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . DB06480 : a review of its use in the management of chronic constipation . The highly selective serotonin Q13639 receptor agonist prucalopride ( Resolor(®) , Resotran(®) , Resotrans(®) ) is indicated for the treatment of chronic constipation . In four randomized , double-blind , multicentre , 12-week trials in patients ( predominantly women ) with chronic constipation , oral prucalopride 2 mg once daily improved bowel function to a significantly greater extent than placebo , with a significantly greater proportion of prucalopride than placebo recipients achieving an average of ≥3 spontaneous , complete bowel movements per week ( primary endpoint ) . Significantly greater improvements in health-related quality of life , patient satisfaction with treatment and bowel habit , and a range of constipation-related symptoms were also seen with prucalopride than with placebo . Satisfaction with treatment and bowel habit was maintained with prucalopride in the longer term . DB06480 was generally well tolerated in patients with chronic constipation , with the most commonly reported adverse events ( headache , nausea , abdominal pain , diarrhoea ) primarily occurring on the first day of treatment . During the clinical trials , no cardiovascular safety issues have arisen in patients with chronic constipation receiving prucalopride . In conclusion , prucalopride is an important option for use in patients with chronic constipation who have not experienced adequate relief with laxatives . 5-HT₄ receptor stimulation leads to soluble AβPPα production through P14780 upregulation . Serotonin 4 ( Q13639 ) receptor signaling does not only have the physiological function of improving cognition , but might also be helpful in the therapy of Alzheimer 's disease ( AD ) through regulation of the production of soluble amyloid-β protein precursor alpha ( sAβPPα ) . To analyze the relationship between Q13639 receptor signaling and sAβPPα production , we stably transfected H4 cells with AβPP and Q13639 receptor ( H4/AβPP/ Q13639 cells ) . We found that 24-h incubation with the Q13639 receptor agonist RS-67333 upregulates matrix metalloproteinase-9 ( P14780 ) . Furthermore , P14780 overexpression enhanced sAβPPα levels , whereas knockdown with P14780 siRNA decreased sAβPPα levels . When RS-67333 was injected for 10 days in Tg2576 mice , a model of amyloid-β peptide ( Aβ ) deposition , there was an increase in hippocampal levels of sAβPPα , C-terminal fragment α , and P14780 , as well as a decrease in hippocampal senile plaque number and levels of the 40 amino acid peptide , Aβ40 . Taken all together , these experiments demonstrate that Q13639 receptor stimulation induces expression of P14780 which cleaves AβPP through α-secretase-like activity , leading to an increase of sAβPPα levels and a reduction of Aβ load . Proteases and lipoprotein receptors in Alzheimer 's disease . Alzheimer 's disease ( AD ) is the leading cause of senile dementia , and is a complex disorder . The pathological hallmarks of AD were discovered by Dr. Alois Alzheimer in 1907 , and include deposits of amyloid or senile plaques and neurofibrillar tangles . Plaques are composed of a peptide , termed the Abeta peptide , that is derived by proteolytic processing of the amyloid precursor protein ( P05067 ) , while neurofibrillar tangles result from a hyperphosphorylation of the tau protein . Mechanisms associated with the formation of plaques and neurofibrillar tangles and their respective contributions to the disease process have been intensely investigated . Proteolytic processing of P05067 that results in the generation of the Abeta peptide is now well understood and is influenced by several proteins . Recent evidence suggests that the Abeta levels are carefully regulated , and several proteases play an important role in removing the Abeta peptide . Finally , it is becoming apparent that several members of the P01130 family play important roles in the brain , and may modulate the course of AD . p-Chloroamphetamine , a serotonin-releasing drug , elicited in rats a hyperglycemia mediated by the P08908 and P41595 /2C receptors . The effects of a serotonin ( 5-HT ) releasing drug , p-chloroamphetamine , on plasma glucose levels were investigated in rats . p-Chloroamphetamine elicited a significant hyperglycemia . The hyperglycemic effects of p-chloroamphetamine were completely prevented by the 5-HT synthesis inhibitor , p-chlorophenylalanine . Prior adrenodemedullation abolished the hyperglycemia elicited by p-chloroamphetamine . p-Chloroamphetamine-induced hyperglycemia was prevented by methysergide , which blocks the 5-HT1 and 5-HT2 receptor , the P08908 /1B/2C receptor antagonist , (-)-propranolol , the selective P08908 receptor antagonist , 4- ( 2'-methoxyphenyl-1- [ 2'-n-2 " pyridinyl ) -p-iodobenzamido ] -ethyl-pi perazine ( p-MPPI ) , the 5- Q13049 /2B/2C receptor antagonists , ritanserin and 4-isopropyl-7-methyl-9-(2-hydroxy-1-methyl-propoxycarbonyl)-4,6A,7 , 8,9,10,10A-octahydro-indolo[4,3-FG]quinolone maleate ( LY 53857 ) . However , the 5- Q9H205 and Q13639 receptor antagonist , tropisetron , the Q13639 receptor antagonist , 2-methoxy-4-amino-5-chloro-benzoic acid 2-(diethylamino) ethyl ester ( SDZ 205-557 ) , and the 5- Q13049 receptor antagonist , ketanserin , did not affect the p-chloroamphetamine-induced hyperglycemia . These results suggest that p-chloroamphetamine-induced hyperglycemia is elicited by an enhanced 5-HT release and facilitated adrenaline release . Moreover , our results indicate that p-chloroamphetamine-induced hyperglycemia is mediated by P08908 and P41595 /2C receptors . Spinal cord injury induces early and persistent lesional Q99571 receptor expression . Following spinal cord injury ( SCI ) , neuropathic , chronic pain is a major cause of disability . Recently , glial Q99571 receptor ( P2X4R ) has been identified as a major contributor to the development of neuropathic pain after peripheral nerve injury . Here we report analysis of P2X4R expression following rat SCI . Significant lesional accumulation of P2X4R+ cells was detected as early as 24 h after SCI , reaching maximum cell numbers on Day 7 . Thereafter cell numbers declined but persisted at significantly elevated , sub-maximal levels ( > 70 % ) until 1 month post injury . Double-immunolabeling identified the majority of lesional P2X4R+ cells as activated microglia/macrophages and surviving neurons/neurites . Increase of P2X4R+ , beta- P05067 + hypertrophic neurites correlated with proximity to the lesion . Further , P2X4R+ cells coexpressed the intracellular regulators of signalling cascades , P23219 ( > 20 % ) , P35354 ( > 5 % ) , RhoA ( > 60 % ) and RhoB ( > 10 % ) . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . 5-hydroxytryptamine and its receptors in systemic vascular walls . 5-hydroxytryptamine ( 5-HT ) in the bloodstream is largely contained in platelets and circulates throughout the entire vascular system . 5-HT released from activated platelets dramatically changes the function of vascular smooth muscle cells ( VSMCs ) and endothelial cells ( ECs ) . In VSMCs , 5-HT induces proliferation and migration via 5- Q13049 receptors . These effects are further enhanced by vasoactive substances such as thromboxane A2 and angiotensin II . 5- Q13049 receptor activation in VSMCs also causes both enhancement of prostaglandin I2 production by inducing cyclooxygenase-2 and reduction of nitric oxide ( NO ) by suppressing inducible NO synthase . Evidence showing that 5-HT in ECs plays a principal role in angiogenesis now exists . Stimulation of 5-HT1 and/or 5-HT2 receptors has been implicated in the angiogenic effect of 5-HT . The extracellular signal-regulated kinase and endothelial NO synthase ( P29474 ) activation-dependent pathways are involved in the mechanisms . Moreover , Q13639 receptors in ECs have been shown to also regulate angiogenesis . Recent reports show sarpogrelate , a selective antagonist of the 5- Q13049 receptor , indirectly enhances the function of P28222 receptors in ECs via inhibition of 5- Q13049 receptors in VSMCs or platelets . This indirect action of P28222 receptors in ECs may increase NO production derived from P29474 and a vasodilator response . Furthermore , sarpogrelate and other 5- Q13049 receptor antagonists have been shown to reduce the constitutive activity of 5- Q13049 receptors . It is believed that increasing evidence on the role of 5-HT receptors will contribute to the expansion of the clinical application of existing therapeutic drugs such as sarpogrelate , and to the development of new 5-HT receptor-related drugs for treating cardiovascular diseases . Evaluation of hypoxia inducible factor expression in inflammatory and neurodegenerative brain models . The neuroinflammatory process is thought to contribute to the progression of neurological disorders and brain pathologies . The release of pro-inflammatory cytokines and chemokines by activated glial cells , astrocytes and microglia plays an important role in this process . However , the role of hypoxia-inducible factor-1α ( HIF-1α ) , the key transcription factor regulating the expression of hypoxia-inducible genes , during glial activation is less known . Thus , we examined the significance of HIF-1α in three experimental models : first in an acute model of inflammation induced by pro-inflammatory cytokines P01375 -α , IL-1β and IFN-γ ; secondly , in a chronic model of inflammation using an APPswe/PS1dE9 ( P05067 / P49768 ) transgenic mouse model of Alzheimer 's disease and thirdly via the inhibition of the PI3K/AKT pathway in a model of neuronal apoptosis . During acute glial inflammation induced by in vitro administration of P01375 -α , IL-1β and IFN-γ , mRNA expression levels of HIF-1α were significantly upregulated ; however , this effect was blocked by SP600126 , a pharmacological inhibitor of mitogen-activated protein kinases ( MAPKs ) . These data suggest that MAPKs could be involved in HIF-1α regulation . In addition , we observed that HIF-1α is not involved in the neuronal apoptotic process mediated by P19957 -kinase inhibition , which is regulated by c-Jun . Finally , we did not detect significant differences in the expression of HIF-1α mRNA in P05067 / P49768 mice during the course of the study ( 3-12 months of age ) . Thus , we demonstrated that HIF-1α has a prominent role in acute but not in chronic inflammatory processes , such as the one which occurs in the P05067 / P49768 experimental model of AD . Moreover , HIF-1α is not involved in neuronal apoptosis after PI3K/AKT inhibition . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . New-generation Q13639 receptor agonists : potential for treatment of gastrointestinal motility disorders . IMPORTANCE OF THE FIELD : Gastrointestinal ( GI ) dysmotility is an important mechanism in functional GI disorders ( FGIDs ) including constipation , irritable bowel syndrome , functional dyspepsia , and gastroparesis . 5-hydroxytryptamine(4) ( 5-HT(4) ) receptors are targets for the treatment of GI motility disorders . However , older 5-HT(4) receptor agonists had limited clinical success because they were associated with changes in the function of the cardiac Q12809 potassium channel . AREAS COVERED IN THIS REVIEW : We conducted a PubMed search using the following key words alone or in combination : 5-HT(4) , safety , toxicity , pharmacokinetics , pharmacodynamics , clinical trial , cardiac , hERG , arrhythmia , potassium current , elderly , prucalopride , ATI-7505 , and velusetrag ( DB05655 ) , to review mechanisms of action , clinical efficacy , safety and tolerability of three new-generation 5-HT(4) receptor agonists . WHAT THE READER WILL GAIN : DB06480 , ATI-7505 , and velusetrag ( DB05655 ) are highly selective , high-affinity 5-HT(4) receptor agonists that are devoid of action on other receptors within their therapeutic range . Their efficacy has been demonstrated in pharmacodynamic studies which demonstrate acceleration of colonic transit and , to a variable degree , in clinical trials that significantly relieve chronic constipation . Currently available evidence shows that the new 5-HT(4) receptor agonists have safe cardiac profiles . TAKE HOME MESSAGE : New-generation 5-HT(4) receptor agonists and future drugs targeting organ-specific splice variants are promising approaches to treat GI dysmotility , particularly colonic diseases . DB06480 for chronic constipation . Chronic constipation is a frequently reported medical disorder that reduces patients ' quality of life and imposes a significant economic burden on the health care system . Symptoms of constipation are diverse and include infrequent bowel movements , hard stool , straining at stool , sensations of anorectal obstruction and feelings of incomplete evacuation . Patients with chronic constipation can be categorized into one of three main groups based on their underlying pathophysiology : normal transit constipation ; colonic inertia ; and pelvic floor dyssynergia . Specialized tests ( i.e. , anorectal manometry , radio-opaque marker study ) may be required in some patients to help distinguish the different subtypes of constipation and to guide appropriate therapy . Although the underlying mechanism of constipation differs among patients , serotonin ( 5-hydroxytryptamine ( 5-HT ) ) appears to have an important role in colonic motility in some patients . Previous research has demonstrated that stimulation of Q13639 receptors improves symptoms of chronic constipation in some patients . DB06480 , a selective Q13639 agonist , relieved symptoms of constipation in phase II and phase III clinical trials . In this monograph , we review the pharmacology , mechanism of action , efficacy and safety of the selective Q13639 agonist prucalopride in patients with chronic constipation . Single-walled carbon nanotubes ( SWCNTs ) enhance DB00761 - , acetylcholine- , and serotonin-induced contractions and evoke oxidative stress on rabbit ileum . We examined the effects of intravenous administration of purified arc-discharge single-walled carbon nanotubes ( SWCNTs ) on rabbit ileum to establish the possibility of using these SWCNTs as cell markers or drug carriers for the treatment of intestinal diseases . The SWCNT purification process eliminated carbonaceous impurities and decreased the amount of metals . SWCNTs increased the contractile responses induced by DB00761 , acetylcholine ( ACh ) , and serotonin ( 5-HT ) in rabbit ileum . Verapamil , apamin , glibenclamide , quinine and charybdotoxin reduced the contractile responses induced by ACh and 5-HT in ileum from rabbits treated with SWCNTs , indicating that voltage-dependent Ca2+ channels and small , intermediate , and large-conductance Ca(2+)-activated , DB00171 -sensitive , and voltage-dependent K+ channels are involved in these effects . Atropine and hexamethonium reduced the ACh response , indicating that muscarinic and nicotinic receptors are involved in this effect . DB00904 and GR 113808 reduced the 5-HT response , indicating that serotonin 5- Q9H205 and Q13639 receptors are involved in this effect . SWCNTs increased the malondialdehyde plus 4-hydroxyalkenals and carbonyl levels in rabbit plasma and ileum , indicating that SWCNTs produce oxidative stress . SWCNTs did not produce relevant histological changes or modify the levels of the inflammatory mediators P35228 and P35354 in the ileum . In conclusion , this study demonstrates that the intravenous administration of SWCNTs can evoke oxidative stress and affect contractility in rabbit ileum . These effects could reduce the possibility of using the arc-discharge SWCNTs as cell markers or drug carriers to treat intestinal diseases .

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[ DB00563 for treatment of rheumatoid arthritis ] . Rheumatoid arthritis ( RA ) is a common chronic inflammatory and destructive arthropathy that can not be cured and that has substantial personal , social and economic costs . DB00563 is a well-known folate analogue , and is the most frequently applied drug for the disease to modify antirheumatic therapy in patients with rheumatoid arthritis . Although results of studies have shown the efficacy of such drugs as methotrexate on rheumatoid arthritis , activity measures and their effect on mortality in patients with the disease remain unknown . The current therapeutic approach to rheumatoid arthritis consists of administration of anti-inflammatory , immunomodulating , and immunosuppressive drugs . Immunomodulating drugs , as opposed to non-steroid anti-inflammatory drugs , which only reduce the signs of inflammation , are capable of gradually checking the course of the disease . However , they do not prevent the slow but progressive destruction of joints . The prominent role of proinflammatory cytokines and growth factors in the pathogenesis of RA has been documented . Recent studies have demonstrated the efficacy of anticytokine treatment . DB00065 is a chimeric monoclonal antibody capable of neutralizing human P01375 alpha . A number of clinical trials for the treatment of rheumatoid arthritis with infliximab indicated that P01375 alpha blockade was effective and well tolerated , with excellent results occurring at 3 and 10 mg/kg in combination with methotrexate . Treatment of RA patients with the combination of infliximab and methotrexate also prevented radiographic evidence of progression of joint damage . If its clinical efficacy is sustainable and its safety confirmed over the long term , infliximab may become an essential agent of choice for the treatment of RA . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . DB00065 treatment for steroid-refractory acute graft-versus-host disease after orthotopic liver transplantation : a case report . Acute graft-versus-host disease ( GVHD ) following orthotopic liver transplantation is a rare but severe disease with a 75 % death rate in adults . Various therapeutic strategies have been proposed for steroid-refractory GVHD , but there is still no consensus . P01375 -alpha is a key inflammatory cytokine involved in acute GVHD physiopathology , and infliximab has shown encouraging results for the treatment of acute GVHD following hematopoietic stem cell transplantation . We report the first case of acute GVHD following liver transplantation that was refractory to steroids and anti-lymphocyte globulin but was successfully treated with infliximab . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . [ DB00065 in the treatment of rheumatoid arthritis ] . DB00065 is a chimeric monoclonal antibody capable of neutralizing human P01375 alpha . A number of clinical trials for the treatment of rheumatoid arthritis(RA) with infliximab indicated that P01375 alpha blockade was effective and well tolerated with the excellent results occurring at 3 and 10 mg/kg in combination with methotrexate . Treatment of RA patients with the combination of infliximab and methotrexate also prevented radiographic evidence of progression of joint damage . If the clinical efficacy is sustained and the safety is confirmed over the long term , infliximab may become an essential agent of choice for the treatment of RA . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . DB00065 for the therapy of chronic sarcoidosis , Baughman RP , Drent M , Kavuru M et al. : DB00065 therapy in patients with chronic sarcoidosis and pulmonary involvement . Am . J . Respir . Crit . Care Med . ( 2006 ) 174(7):795-802 . Sarcoidosis is an inflammatory multiorgan disease in which the lungs are the most commonly affected . It can also involve the skin , lupus pernio being a common form of chronic cutaneous sarcoidosis . The histopathologically specific lesion is represented by non-caseating granulomas occurring in the involved organs , with P01375 playing a role in granuloma generation . Several therapies are available , with corticosteroids representing the conventional therapy given as topic or systemic formulations . Anti- P01375 therapies ( such as etanercept or infliximab ) have been assessed so far , the latter most commonly in refractory sarcoidosis . The discussed study evaluates the safety and efficacy of infliximab in chronic sarcoidosis with pulmonary manifestations . DB00065 restores glucose homeostasis in an animal model of diet-induced obesity and diabetes . P01375 plays an important role in obesity-linked insulin resistance and diabetes mellitus by activating at least two serine kinases capable of promoting negative regulation of key elements of the insulin signaling pathway . Pharmacological inhibition of P01375 is currently in use for the treatment of rheumatoid and psoriatic arthritis , and some case reports have shown clinical improvement of diabetes in patients treated with the P01375 blocking monoclonal antibody infliximab . The objective of this study was to evaluate the effect of infliximab on glucose homeostasis and insulin signal transduction in an animal model of diabetes . Diabetes was induced in Swiss mice by a fat-rich diet . DB09341 and insulin homeostasis were evaluated by glucose and insulin tolerance tests and by the hyperinsulinemic-euglycemic clamp . Signal transduction was evaluated by immunoprecipitation and immunoblotting assays . Short-term treatment with infliximab rapidly reduced blood glucose and insulin levels and glucose and insulin areas under the curve during a glucose tolerance test . Furthermore , infliximab increased the glucose decay constant during an insulin tolerance test and promoted a significant increase in glucose infusion rate during a hyperinsulinemic-euglycemic clamp . In addition , the clinical outcomes were accompanied by improved insulin signal transduction in muscle , liver , and hypothalamus , as determined by the evaluation of insulin-induced insulin receptor , insulin receptor substrate-1 , and receptor substrate-2 tyrosine phosphorylation and Akt and forkhead box protein O1 serine phosphorylation . Thus , pharmacological inhibition of P01375 may be an attractive approach to treat severely insulin-resistant patients with type 2 diabetes mellitus . Acute renal artery occlusion following infliximab infusion . We report the case of a 44-year-old male patient who presented with acute renal artery occlusion , 3 d after first injection of infliximab for steroid refractory attack of ulcerative colitis . Extensive work-up provided no evidence of predisposing factors for arterial thrombosis . DB00065 was thus suspected in the genesis of thrombosis , based on both intrinsic and extrinsic criteria . At month 3 after thrombosis with ongoing anticoagulation , angio-tomodensitometry showed complete revascularization of the left renal artery with renal atrophy . Renal function remained normal and the patient was still in steroid free remission on mercaptopurin monotherapy at maximal follow-up . Few thromboembolic events have been described with anti- tumor necrosis factor ( P01375 ) agents , but it is the first case reported of renal artery thrombosis after infliximab infusion . In addition , we review thrombosis associated with anti- P01375 agents . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Anti- P01375 alpha therapy of rheumatoid arthritis : what have we learned ? Rheumatoid arthritis ( RA ) , a systemic disease , is characterized by a chronic inflammatory reaction in the synovium of joints and is associated with degeneration of cartilage and erosion of juxta-articular bone . Many pro-inflammatory cytokines including P01375 alpha , chemokines , and growth factors are expressed in diseased joints . The rationale that P01375 alpha played a central role in regulating these molecules , and their pathophysiological potential , was initially provided by the demonstration that anti- P01375 alpha antibodies added to in vitro cultures of a representative population of cells derived from diseased joints inhibited the spontaneous production of IL-1 and other pro-inflammatory cytokines . Systemic administration of anti- P01375 alpha antibody or sTNFR fusion protein to mouse models of RA was shown to be anti-inflammatory and joint protective . Clinical investigations in which the activity of P01375 alpha in RA patients was blocked with intravenously administered infliximab , a chimeric anti- P01375 alpha monoclonal antibody ( mAB ) , has provided evidence that P01375 regulates P05231 , P10145 , P13500 , and P15692 production , recruitment of immune and inflammatory cells into joints , angiogenesis , and reduction of blood levels of matrix metalloproteinases-1 and -3 . Randomized , placebo-controlled , multi-center clinical trials of human P01375 alpha inhibitors have demonstrated their consistent and remarkable efficacy in controlling signs and symptoms , with a favorable safety profile , in approximately two thirds of patients for up to 2 years , and their ability to retard joint damage . DB00065 ( a mAB ) , and etanercept ( a sTNF-R-Fc fusion protein ) have been approved by regulatory authorities in the United States and Europe for treating RA , and they represent a significant new addition to available therapeutic options . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . DB00065 's influence on anastomotic strength and degree of inflammation in intestinal surgery in a rabbit model . BACKGROUND : DB00065 , a P01375 -α inhibitor , is a potent anti-inflammatory drug in the treatment of inflammatory bowel diseases . Recent studies have investigated the effect of infliximab treatment on postoperative complications such as anastomotic leakage , however , with conflicting results and conclusions . The purpose of this study was to investigate whether a single dose infliximab has an adverse effect on the anastomotic healing process , observed as reduced anastomotic breaking strength and histopathologically verified lower grade of inflammatory response , in the small intestine of a rabbit . METHODS : Thirty New Zealand rabbits ( median weight 2.5 kg ) were allocated to treatment with an intravenous bolus of either 10 mg/kg infliximab ( n = 15 ) or placebo ( n = 15 ) . One week later all rabbits underwent two separate end-to-end anastomoses in the jejunum under general anesthesia . At postoperative day three , the anastomotic breaking strength was determined and histopathological changes were examined . RESULTS : The mean value of anastomotic breaking strength in the placebo group was 1.89 ± 0.36 N and the corresponding value was 1.81 ± 0.33 N in the infliximab treated rabbits . There was no statistically significant difference between the groups ( p = 0.51 ) . The infliximab-treated rabbits had a significant lower degree of inflammatory infiltration response compared to the placebo group ( p = 0.047 ) . CONCLUSIONS : Our conclusion , limited by the small sample sizes in both groups , is that a single dose of infliximab , given one week prior to surgery , does not have an impact on the anastomotic breaking strength on the third postoperative day in the small intestine of rabbits . A clinical study assessing the tolerability and biological effects of infliximab , a P01375 inhibitor , in patients with advanced cancer . BACKGROUND : Tumour necrosis factor-alpha ( P01375 ) is an important regulator of the chronic inflammation contributing to tumour progression . DB00065 , an anti- P01375 monoclonal antibody was investigated in this trial of patients with advanced cancer . The primary objectives were to determine the safety profile and biological response of infliximab in a cancer population . Clinical response was a secondary objective . PATIENTS AND METHODS : Forty-one patients received infliximab at 5 mg/kg ( n = 21 ) or 10 mg/kg ( n = 20 ) i.v. at 0 and 2 weeks and then every 4 weeks . Post-treatment samples were measured for changes in plasma and serum P01375 , P13500 , P05231 and P02741 ( CRP ) . RESULTS : DB00065 was well tolerated with no dose-limiting toxic effects . At both doses of infliximab , neutralisation of serum P01375 was observed after 1 h while plasma P13500 , P05231 and serum CRP were decreased 24 and 48 h following infliximab administration . Seven patients experienced disease stablisation ( range 10-50+ weeks ) . There was no evidence of disease acceleration in any patient . CONCLUSIONS : DB00065 treatment was safe and well tolerated in patients with advanced cancer . There was evidence of biological activity with baseline P01375 and P13500 being correlated with infliximab response . Tumour necrosis factor-alpha blockade : a new era for effective management of rheumatoid arthritis . Tumour necrosis factor ( P01375 ) -alpha inhibitors have emerged as a new treatment option for rheumatoid arthritis ( RA ) . The scientific rationale for targeting P01375 in RA derives from extensive work in the laboratory , showing the importance of this pro-inflammatory cytokine as a mediator of joint inflammation . Proof of principle has now been firmly established in clinical trials where P01375 inhibitors have been shown to decrease the signs and symptoms of joint inflammation and slow radiological progression of joint damage . Presently , the two P01375 inhibitors available for use in RA are etanercept and infliximab . DB00005 is a soluble P01375 receptor : Fc fusion protein that competes with the endogenous P01375 receptors for P01375 binding . DB00065 is a chimeric anti- P01375 monoclonal antibody , which also binds with high affinity to soluble P01375 . DB00005 and infliximab will be rapidly incorporated into current treatment paradigms , which call for early and intensive treatment of RA using disease-modifying antirheumatic drugs ( DMARDs ) , such as methotrexate , sulfasalazine and hydroxychloroquine . A major drawback to the widespread use of these biologics is their high costs . Some patients with limited financial means may be denied access to these effective anti-inflammatory agents . Moreover , long-term experience with P01375 inhibitor therapy has been limited and concerns linger about the possibility that etanercept and infliximab may cause unforeseen side effects or increase the risk for opportunistic infection . Despite these caveats , P01375 inhibitors represent a major advance for the treatment of RA and will likely spawn new indications for anti- P01375 therapy and the development of novel therapeutic compounds with similar biological activity . Efficacy of infliximab in a patient with refractory idiopathic retroperitoneal fibrosis . Glucocorticoids are the mainstay of treatment of idiopathic retroperitoneal fibrosis ( Q969Q1 ) . However , relapses are frequent upon tapering of the glucocorticoid dose . A variety of traditional immunosuppressants have been proposed as steroid-sparing agents , but some patients fail to adequately respond to combined glucocorticoid and immunosuppressive therapy . We report a patient with Q969Q1 refractory to combined glucocorticoid and methotrexate therapy treated with the anti- P01375 -α monoclonal antibody infliximab . DB00065 was administered at 5 mg/kg/bodyweight at week 0 , 2 , 6 and 8-weekly thereafter for 3 consecutive years . Drug efficacy and safety were assessed clinically and by laboratory tests at treatment onset and subsequently before each infusion . In addition , 18FFluorodeoxyglucose ( DB09150 ) positron emission computerised tomography ( PET/CT ) and abdominal CT scans were used to monitor disease activity and response to treatment . DB00065 therapy resulted in a satisfactory clinical and laboratory response paralleled by an improvement in imaging findings . No serious adverse events were noted . DB00065 may be an effective and safe treatment for refractory Q969Q1 . A controlled study is required to confirm our findings . Development of interstitial pneumonia in a rheumatoid arthritis patient treated with infliximab , an anti-tumor necrosis factor alpha-neutralizing antibody . DB00065 , an anti-tumor necrosis factor ( P01375 ) -alpha antibody , was introduced to a 66-year-old woman with methotrexate ( MTX ) -resistant rheumatoid arthritis ( RA ) . Although the P01375 -blocking therapy was successful , she developed noninfectious interstitial pneumonia ( IP ) after a second infusion of infliximab . In most cases reported previously , infliximab-associated noninfectious IP occurred after a second or third infusion of infliximab , and this type of IP was more fatal in comparison with cases associated with MTX treatment alone . Keeping a sharp lookout on IP development during this period is crucial to the success of infliximab treatment . After MTX discontinuation and steroid pulse therapy , our patient made a dramatic recovery from IP . [ Treatment of Crohn disease in adults with tumor necrosis factor-alpha ( P01375 ) antibodies ] . Crohn 's disease ( CD ) is a chronic inflammatory disease of the bowel characterized by segmental transmural inflammation and granulomatous changes . P01375 alpha is a member of a large family of proteins and receptors that are involved in immune regulation . It is a proinflammatory and immunoregulatory cytokine synthesized by monocytes , macrophages , and T cells . P01375 alpha plays an early central role in the cytokine cascade of the inflammatory process . Recently , chimeric monoclonal antibodies that inhibits P01375 alpha have been used in the treatment of Crohn 's disease . DB00065 has been the most largely used antibody . It is commercialized in the USA and has recently obtained an European marketing approvement . DB00065 is indicated for the treatment of moderately to severely active CD in patients having an inadequate response to conventional therapy . Clinical trials have demonstrated efficacy when the agent is initiated as a 5 mg/kg single intravenous infusion . In patients with fistulizing CD , administration of 2 subsequent 5 mg/kg doses 2 and 6 weeks after the initial dose appears to be efficacious . Limited clinical data also suggest that infliximab retreatment regimen restores response and maintains remission rates . DB00065 appears to be well tolerated . To date , very little is known about the potential for long-term toxicity with infliximab therapy . DB00065 -Induced Hypothyroidism : A Novel Case and Postulations concerning the Mechanism . We report a patient with cutaneous sarcoidosis who developed hypothyroidism following 17 months of infliximab therapy . To our knowledge , this is the first reported case of hypothyroidism following infliximab administration . While it is possible that the patient 's hypothyroidism was unrelated to the use of infliximab , the time course and lack of alternative explanations make such an association plausible . We postulate that hypothyroidism in this patient may have been related to the development of autoantibodies to infliximab that triggered the development of an autoimmune thyroiditis . Regardless of the mechanism , we would encourage clinicians to keep the potential mechanisms of P01375 -α in mind when treating patients with P01375 -α antagonist medications . DB00065 as rescue medication for patients with severe ulcerative/indeterminate colitis refractory to tacrolimus . BACKGROUND : The calcineurin inhibitor tacrolimus and the anti- P01375 -antibody infliximab are established options in steroid-refractory ulcerative colitis ( UC ) . AIM : To evaluate the efficacy of infliximab-salvage therapy in patients with refractory UC failing to respond to tacrolimus . METHODS : Twenty-four patients were enrolled in this evaluation . Reasons for tacrolimus therapy were steroid-refractory disease in 19 patients and steroid-dependency in five patients . All patients receiving infliximab had tacrolimus-refractory active disease ( Lichtiger score > 10 ) and were treated with 5 mg/kg at weeks 0 , 2 and 6 and every 8 weeks thereafter , if tolerated . RESULTS : Six of 24 patients ( 25 % ) achieved remission following infliximab infusion and four of 24 ( 17 % ) had an initial response only , but underwent proctocolectomy later because of loss of response ( 3 ) or development of a delayed hypersensitivity reaction ( 1 ) . Fourteen patients ( 58 % ) completely failed to respond with 10 undergoing colectomy . Eight patients experienced side effects under infliximab , including two infectious complications ( herpes zoster and herpes pneumonia ) . CONCLUSIONS : DB00065 offers a therapeutic option as rescue therapy in about a quarter of patients with active UC after failing to respond to tacrolimus . This benefit has to be weighed against the risks of infectious complications . DB00065 in patients who have spondyloarthropathy : clinical efficacy , safety , and biological immunomodulation . A major breakthrough has been achieved in the treatment of patients who have AS and other types of SpA . The identification of the expression and role of P01375 in patients who have these diseases and the recognition of their relation with gut inflammation ( where infliximab therapy has proven efficacious already ) has led to the successful use of P01375 blockade in SpA , establishing a new indication for this type of anticytokine therapy . Evidence supports equal response in cases of axial or peripheral disease . DB00065 therapy has been most extensively documented in this new indication for anti- P01375 therapy , but other compounds are also in the field . Gorman et al reported on 40 patients who had active AS who were randomly assigned to receive twice-weekly subcutaneous injections of etanercept ( 25 mg ) or placebo for 4 months [ 65 ] . The primary endpoint was a composite of improvements . Treatment with etanercept resulted in significant and sustained improvement ( treatment response in 80 % in the etanercept group versus 30 % in the placebo ) . Data regarding the human anti- P01375 monoclonal antibody adalimumab in SpA are not yet available . Different questions remain open , including optimal dosing , long-term safety , and effects of this new treatment on the structural articular level ; however , a therapeutic breakthrough like the one currently reviewed has seldom occurred in arthritis care . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Tumour necrosis factor in sarcoidosis and its potential for targeted therapy . Tumour necrosis factor ( P01375 ) -alpha is a potent cytokine involved in the inflammatory reactions of many acute and chronic diseases . Recently , agents that block TNFalpha either directly or indirectly have been successful in the treatment of a variety of immune-mediated inflammatory disorders including rheumatoid arthritis and Crohn 's disease . Sarcoidosis is an immune-mediated inflammatory disorder characterised by the formation of granulomas . TNFalpha is important in the initiation and perpetuation of inflammation in sarcoidosis , contributing to the initiation of granulomas and the progression of fibrosis , as well as to nongranulomatous inflammation . Various agents used to treat sarcoidosis affect P01375 , including the most widely used drug class , corticosteroids , which are usually effective in blocking TNFalpha release from cells . Other agents that nonspecifically inhibit TNFalpha release include methotrexate , azathioprine and pentoxifylline . Specific P01375 -antagonising biological agents such as infliximab and etanercept are being tested in patients with sarcoidosis , with mixed success . DB00065 has been shown to produce clinical improvement and reduce the requirement for corticosteroids in a small number of patients with sarcoidosis . However , as infliximab can be associated with reactivation of tuberculosis , which could be mistaken as worsening sarcoidosis , it should be used with caution in this patient group . DB00065 : a new therapeutic agent in acute pancreatitis ? PURPOSE : P01375 alpha ( P01375 ) has a central role in the pathogenesis of acute pancreatitis and related systemic complications . The aim of this study is to investigate the therapeutic effectiveness of monoclonal P01375 antibody ( infliximab ) in acute edematous and severe necrotizing pancreatitis models in rats . METHODS : One hundred rats were randomly divided into 10 groups . Acute edematous pancreatitis ( AEP ) was induced by injection of cerulein 20 microg/kg 4 times subcutaneously at hourly intervals . Severe necrotizing pancreatitis ( SNP ) was induced by retrograde injection of 3 % taurocholate into the common biliopancreatic duct . DB00065 8 mg/kg was given via intravenous infusion . Serum amylase activity , pancreatic histopathology , myeloperoxidase enzyme activity ( P05164 ) , and pulmonary changes were assessed . RESULTS : DB00065 treatment significantly decreased serum amylase activity ( 11939 +/- 1914 U/L versus 3458 +/- 915 U/L , P < 0.001 ) and histopathologic score ( 4.1 +/- 0.5 versus 1.5 +/- 0.3 , P < 0.001 ) in AEP . It also suppressed neutrophil infiltration and P05164 activity of the pancreatic tissue . In SNP , infliximab treatment was found to decrease pathologic score ( 9.4 +/- 1.2 versus 3.6 +/- 0.8 , P < 0.001 ) and serum amylase activity ( 20442 +/- 2375 versus 8990 +/- 1730 , P < 0.01 ) . It ameliorated both parenchymal and fatty tissue necrosis of the pancreas . DB00065 also alleviated alveolar edema and acute respiratory distress syndrome like pulmonary complications , but the difference was not significant . CONCLUSIONS : Chimeric P01375 antibody , infliximab , should be evaluated for treatment of acute pancreatitis . Attenuation of experimental autoimmune myocarditis by blocking activated T cells through inducible costimulatory molecule pathway . OBJECTIVE : Inducible costimulator ( Q9Y6W8 ) is a member of the P10747 family . Although inflammation is an essential pathological feature of myocarditis , the role of Q9Y6W8 in myocarditis remains unclear . METHODS AND RESULTS : Lewis rats were immunized on day 0 with purified porcine cardiac myosin to establish experimental autoimmune myocarditis ( EAM ) . Flow cytometry was used to examine expression of Q9Y6W8 on myocardial infiltrating cells . Anti- Q9Y6W8 antibody or Q9Y6W8 -immunoglobulin ( ICOSIg ) was administered intravenously , and rats were killed on day 14 or 21 to study effects of Q9Y6W8 / Q9Y6W8 -ligand ( O75144 ) pathway blockade during the antigen priming phase ( days 0-14 ) or immune response phase ( days 14-21 ) , respectively . The heart weight to body weight ratio was determined , and histological examination and echocardiogram were performed to evaluate the severity of the disease . Cytokine expression in the heart and T cell proliferation against cardiac myosin were analyzed . Flow cytometry revealed that the majority of infiltrating cells , especially P01730 -positive cells , expressed Q9Y6W8 . Blockade of the Q9Y6W8 / O75144 pathway during the immune response phase attenuated EAM development . However , blockade of the Q9Y6W8 / O75144 pathway during the antigen priming phase did not attenuate and exacerbate EAM . Blockade of T cell activation through Q9Y6W8 suppressed expression of cytokines including P27352 -gamma , P05112 , P05231 , P22301 , P01584 , and P01375 and inhibited T cell proliferation in vitro . CONCLUSIONS : Blockade of T cell activation through Q9Y6W8 during the immune response phase regulates development of EAM , and therefore , Q9Y6W8 may be an effective target for treating myocarditis . DB00065 in relapsing polychondritis . Relapsing polychondritis ( RP ) is a rare systemic disease of unknown etiology , characterized by recurrent inflammation of cartilaginous structures and other connective tissues , including the ears , nose , joints , respiratory tract , and others . Due to the presence of typical signs and symptoms , biopsy is seldom necessary . Treatment includes corticosteroids , occasionally associated with immunosuppressive agents , but refractory cases are described . Recent reports suggest that anti- P01375 agents , such as infliximab , may be of value in patients who do not respond to conventional therapy , but experience with this treatment is scarce . In this paper , the authors report the case of a patient with RP refractory to combined treatment with corticosteroids and immunosuppressive agents , who showed a good response to infliximab . Effects of infliximab on bacterial translocation in experimental acute necrotizing pancreatitis . BACKGROUND & OBJECTIVES : Translocation of bacteria from the gut is an important factor in the development of septic complications and mortality in acute pancreatitis ( AP ) . The present study was designed to assess the effects of infliximab treatment on bacterial translocation ( BT ) in experimental acute necrotizing pancreatitis . METHODS : Male Sprague-Dawley rats ( n=45 ) were allocated into three groups . AP was induced in group II ( positive control , n=15 ) and group III ( DB00065 ; n=15 ) by retrograde injection of taurocholate into the common biliopancreatic duct . Group I rats ( Sham ; n=15 ) received normal saline infusion into the common biliopancreatic duct as placebo . Groups I and II were treated by normal saline and group III was treated with infliximab intraperitoneally on 6 , 30 and 54 h after induction of pancreatitis . All surviving animals were killed 60 h after the induction of pancreatitis , and specimens were collected for amylase measurement as well as histopathologic and microbiologic examinations . RESULTS : Oedema , acinar cell necrosis , inflammatory infiltration , haemorrhage , fat necrosis and perivascular inflammation in group III rats were decreased with infliximab treatment when compared with group II ( P < 0.001 ) . BT to mesentery lymph node in groups I , II and III were 20 , 100 and 46 per cent , respectively . BT to peritoneum and pancreas in group III was lower than group II ( P < 0.05 ) . INTERPRETATION & CONCLUSIONS : DB00065 administration resulted in beneficial effects on BT and histopathologic changes in the experimental necrotizing pancreatitis . Whether anti- P01375 therapy has a role in prevention of complications of P01160 needs to be established . Incidence and clinical significance of immunogenicity to infliximab in Crohn 's disease : a critical systematic review . BACKGROUND : DB00065 ( IFX ) is a chimeric ( mouse/human ) anti- P01375 monoclonal antibody approved for the treatment of refractory luminal and fistulizing Crohn 's disease ( CD ) . It is a source of potential immunogenicity for humans , with the occurrence of anti-infliximab antibodies ( ATIs ) , which are thought to interfere with the pharmacodynamics and/or pharmacokinetics of the compound . It remains unclear whether ATIs have any clinical importance for drug efficacy or safety . We review studies specifically evaluating the incidence of ATIs in CD and their impact on the efficacy and safety of IFX . METHODS : A systematic review was undertaken by electronic searches of the PubMed and SCOPUS databases from earliest records to October 2008 , as well as reference lists of all relevant articles and relevant abstracts from meetings . RESULTS : The biological and clinical mechanisms of ATI development are poorly understood . The incidence of ATIs in vivo depends on multiple analytical and clinical factors , both patient- and treatment-related . The presence of ATIs is weakly and variably associated with clinical response or infusion reactions , but not with reactions relevant to clinical decision-making . Enormous variation in the methods of reporting ATIs and immunogenicity of IFX make almost any interpretation possible from different studies , but few have clinical relevance . CONCLUSIONS : There is no clear evidence that ATIs have an impact on efficacy or safety , nor a need to measure or prevent them in clinical practice . Circulating drug concentration may be a more relevant measure of immunogenicity . Acute effects of sarpogrelate , a 5- Q13049 receptor antagonist on cytokine production in endotoxin shock model of rats . Serotonin ( 5-HT ) (2A) receptors are involved in cytokine production in infection or sepsis . Therefore , 5-HT(2A) receptor antagonist might be useful to treat sepsis . The present study investigates the effects of a 5-HT(2A) receptor antagonist , sarpogrelate on endotoxin shock . Catheters were inserted into the femoral artery and vein of Sprague-Dawley rats . First , sarpogrelate 0 ( control ) , 3 , or 10 mg/kg dissolved in 0.5 ml of distilled water has been given , followed by endotoxin 10 mg/kg in saline 0.5 ml 5 min later . Blood pressure , pulse rate and survival rate were monitored in 20 rats per dose . Blood gas and plasma cytokine concentrations were measured in 8 rats per dose . In four rats each of sarpogrelate 0 , 3 , or 10 mg/kg , and sham operation , the lung histology was examined . Zero , 15 , and 12 rats survived for 8 h in the control , 3 mg/kg , and 10 mg/kg groups , respectively . The control group had the lowest blood pressure , pulse rate , pH and arterial oxygen tension , and the highest arterial carbon dioxide tension and plasma IL-1beta concentration . The increase of P01375 was significantly lower in 3 mg/kg group than in the control group . Pathological changes of the lung were inhibited in 3 and 10 mg/kg groups . In conclusion , sarpogrelate might be effective to decrease production of pro-inflammatory cytokines , to keep hemodynamics , to inhibit lung damage , and to decrease mortality in endotoxin shock . DB00065 and etanercept are equally effective in reducing enterocyte APOPTOSIS in experimental colitis . Loss of epithelial barrier integrity is considered an early step in the pathogenesis of Crohn 's disease ( CD ) , and the rate of enterocyte apoptosis is one of the determinants of the intestinal barrier function . P01375 -alpha ( P01375 ) , one of the major proinflammatory mediators in CD , is one of the extrinsic signals which initiate apoptosis of enterocytes . The aim of this study was to investigate the early effects of experimental colitis on enterocyte apoptosis , and the effects of two anti- P01375 treatments , infliximab ( IFX ) and etanercept ( ETC ) . In addition , the importance of receptor I for P01375 was tested in TNFR-1 ( -/- ) mice . Circulating P01375 levels were effectively reduced by IFX and ETC ( p < 0.01 , both ) at 3 and 6 h . Apoptosis of the ileal enterocytes , assessed by TUNEL staining , staining for Fas-ligand , and bax , increased at 3 and 6h . These alterations were prevented by both anti- P01375 strategies , and in TNFR-1(-/-) animals . The anti-apoptotic protein Bcl-2 was expressed in the ileal epithelium under control conditions , but was suppressed in DNB-colitis . Expression of Bcl-2 was maintained in both anti- P01375 treatments and TNFR-1(-/-) mice.DNB colitis induced a very early , rapid increase of enterocyte apoptosis . Both anti- P01375 strategies , IFX and ETC , were equally effective in suppressing enterocyte apoptosis , most likely by inactivation of circulating P01375 . [ Safety in the diagnosis and treatment of inflammatory bowel disease ] . The presentations at Digestive Disease Week 2013 emphasized treatment safety . Anti-tumor necrosis factor ( P01375 ) agents and thiopurines are reasonably safe in breastfeeding and pregnancy . Several studies indicate that controlling the risk of tuberculosis when anti- P01375 agents are planned presents several problems , both in the initial diagnosis of latent tuberculosis and in subsequent patient follow-up , given that cases of tuberculosis continue to occur , despite recommendations . Thiopurines increase the risk of lymphoma , but there is no residual risk when these drugs are withdrawn . Despite increasing knowledge of the risks and recommendations on how to avoid them , there remain considerable shortfalls in the application of preventive measures and , more specifically , in vaccinations . DB00065 and cyclosporin produce similar results when used to treat severe outbreaks of ulcerative colitis . Thromboembolism prevention continues to be deficient , and the barriers to effective prevention concern not only physicians but can also involve nursing staff , for example . There is still a wide margin for improvement in safety . New drugs under study ( vedolizumab , DB06674 ) have not shown any hitherto unknown signs of significant toxicity . [ DB00065 in inflammatory bowel diseases -- conference summary and suggested guidelines ] . DB00065 , the monoclonal anti-tumor necrosis factor-alpha ( P01375 ) antibodies preparation , is efficacious in the treatment of inflammatory bowel diseases . However , the optimal therapeutic approach is still under investigation . Reports on side effects and potential complications of infliximab therapy , as well as of other anti- P01375 blocking agents are accumulating . Hence , the Israeli Gastroenterological Association had initiated a conference in order to discuss the frequent clinical issues that have arisen following the use of infliximab for the treatment of inflammatory bowel diseases . The aim was to report on the published clinical experience and problems regarding several practical aspects of the use of DB00065 , to suggest guidelines that are evidence-based and to discuss them with experienced Q9UKU7 -oriented gastroenterologists . The subjects that were discussed include : ( 1 ) treatment protocols ; ( 2 ) maintenance therapy in Crohn 's disease ; ( 3 ) prevention of infections and ( 4 ) therapeutic potential in ulcerative colitis . These topics reflect everyday issues that gastroenterologists deal with while treating inflammatory bowel disease patients . The manuscript summarizes the literature and evidence that were presented in the conference , the points raised at the discussions as well as guidelines suggested by work groups that were established for each subject . These guidelines may assist and direct the gastroenterologist treating inflammatory bowel disease patients with infliximab . DB00065 but not etanercept induces apoptosis in lamina propria T-lymphocytes from patients with Crohn 's disease . BACKGROUND & AIMS : Steroid-refractory Crohn 's disease responds to therapy with the chimeric anti-tumor necrosis factor ( P01375 ) -alpha antibody infliximab . DB00005 , a recombinant P01375 receptor/immunoglobulin G fusion protein , is highly effective in rheumatoid arthritis but not in Crohn 's disease . Because both infliximab and etanercept are P01375 -neutralizing drugs , we investigated the differences in P01375 -neutralizing capacity and human lymphocyte binding and apoptosis-inducing capacity of both molecules . METHODS : We used a nuclear factor kappaB reporter assay and a cytotoxicity bioassay to study P01375 neutralization by infliximab and etanercept . Lymphocyte binding and apoptosis-inducing capacity was investigated using fluorescence-activated cell sorter analysis , annexin V staining , and cleaved caspase-3 immunoblotting using mixed lymphocyte reaction-stimulated peripheral blood lymphocytes ( PBL ) from healthy volunteers and lamina propria T cells from patients with Crohn 's disease . RESULTS : Both infliximab and etanercept neutralized P01375 effectively . DB00065 bound to activated PBL and lamina propria T cells , whereas binding of etanercept was equal to a nonspecific control antibody . DB00065 but not etanercept induced peripheral and lamina propria lymphocyte apoptosis when compared with a control antibody . DB00065 activated caspase 3 in a time-dependent manner , whereas etanercept did not . CONCLUSIONS : Although both infliximab and etanercept showed powerful P01375 neutralization , only infliximab was able to bind to PBL and lamina propria T cells and subsequently to induce apoptosis of activated lymphocytes . These data may provide a biological basis for the difference in efficacy of the 2 P01375 -neutralizing drugs . DB00065 in chronic ocular inflammation . OBJECTIVE : DB00065 is a chimeric antibody which binds tumor necrosis factor ( P01375 ) . It is effective in several chronic inflammatory conditions , including sarcoidosis . METHODS : We report our experience with infliximab in chronic ocular inflammation as part of a retrospective review of all patients treated for chronic inflammatory ocular conditions seen over a 2-year period at our institution . RESULTS : 14 patients with various underlying ocular conditions were treated during the previous two years including patients with sarcoidosis ( 7 ) , Crohn 's disease ( 2 ) , birdshot choroiditis ( 2 ) , idiopathic disease ( 2 ) , Volt-Koyanagi-Harada ( 1 ) and Behçet 's disease ( 1 ) . All patients had persistent inflammation despite systemic immunosuppressive agents and all but one patient experienced marked improvement in ocular inflammation with infliximab . One patient was non-compliant and non-evaluable ; four patients , who had previously received etanercept with either no response ( 3 patients ) or subsequent relapse ( 1 patient ) , responded to infliximab . CONCLUSION : DB00065 is an effective therapy in chronic inflammatory eye disease , especially when related to sarcoidosis . Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced P28906 + cell-derived human dendritic cells . The efficient genetic modification of P28906 + cell-derived dendritic cells ( DC ) will provide a significant advancement towards the development of immunotherapy protocols for cancer , autoimmune disorders and infectious diseases . Recent reports have described the transduction of P28906 + cells via retrovirus- and lentivirus-based gene transfer vectors and subsequent differentiation into functional DC . Since there is significant apprehension regarding the clinical uses of HIV-based vectors , in this report , we compare a murine leukemia virus ( MLV ) - and a human immunodeficiency virus ( HIV ) -based bicistronic vector for gene transfer into human P28906 + cells and subsequent differentiation into mature DC . Each vector expressed both EGFP and the dominant selectable marker P00374 (L22Y) allowing for the enrichment of marked cells in the presence of the antifolate drug trimetrexate ( TMTX ) . Both MLV-based and HIV-based vectors efficiently transduced cytokine mobilized human peripheral blood P28906 + cells . However , in vitro expansion and differentiation in the presence of GM- P04141 , P01375 , Flt-3L , P21583 and P05112 resulted in a reduction in the percentage of DC expressing the transgene . Selection with TMTX during differentiation increased the percentage of marked DC , resulting in up to 79 % ( MLV vector ) and up to 94 % ( lentivirus-vector ) transduced cells expressing EGFP without loss of DC phenotype . Thus , MLV-based vectors and in vitro selection of transduced human DC show great promise for immunotherapy protocols . Cordycepin suppresses P01375 -α-induced NF-κB activation by reducing p65 transcriptional activity , inhibiting IκBα phosphorylation , and blocking IKKγ ubiquitination . Cordycepin is reported to participate in multiple pharmacological activities including anti-tumor and anti-inflammation , and is involved in the regulation of NF-κB signaling pathway . However , the detailed molecular mechanism of cordycepin in suppression of NF-κB signaling pathway remains ambiguous . In this study , we first analyzed the effect of cordycepin on NF-κB activity in P29320 -293T cells , and found that cordycepin resulted in a dose-dependent reduction in P01375 -α-induced NF-κB activation . Although cordycepin did not block P01375 -α-induced nuclear translocation of p65 , high concentration of cordycepin reduced the DNA-binding and transcriptional activities of NF-κB . Moreover , cordycepin also inhibited IκBα phosphorylation so as to suppress the degradation of IκBα . Further investigation revealed that cordycepin suppressed IKKs-mediated NF-κB activation and inhibited the ubiquitination of IKKγ . In conclusion , cordycepin effectively inhibits NF-κB signaling through suppressing the activities of NF-κB , IκB and IKK . Thus , cordycepin may provide some potential therapeutic application in inflammation-associated disorders and cancer . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Defective expression of Q96EB6 contributes to sustain inflammatory pathways in the gut . In inflammatory bowel disease ( Q9UKU7 ) , tissue damage is driven by an excessive immune response , poorly controlled by counter-regulatory mechanisms . Q96EB6 , a class III NAD+-dependent deacetylase , regulates negatively the expression of various proteins involved in the control of immune-inflammatory pathways , such as Stat3 , O15105 , and NF-κB . Here we examined the expression , regulation , and function of Q96EB6 in Q9UKU7 . Q96EB6 RNA and protein expression was less pronounced in whole biopsies and lamina propria mononuclear cells ( LPMCs ) of Q9UKU7 patients in comparison with normal controls . Q96EB6 expression was downregulated in control LPMC by tumor necrosis factor ( P01375 ) -α and interleukin ( IL ) -21 , and upregulated in Q9UKU7 LPMC by neutralizing P01375 -α and IL-21antibodies . Consistently , Q96EB6 expression was increased in mucosal samples taken from Q9UKU7 patients successfully treated with DB00065 . Treatment of Q9UKU7 LPMC with Cay10591 , a specific Q96EB6 activator , reduced NF-κB activation and inhibited inflammatory cytokine synthesis , whereas Ex527 , an inhibitor of Q96EB6 , increased interferon ( IFN ) -γ in control LPMC . Q96EB6 was also reduced in mice with colitis induced by 2,4,6-trinitrobenzenesulphonic acid or oxazolone . Cay10591 prevented and cured experimental colitis whereas Ex527 exacerbated disease by modulating T cell-derived cytokine response . Data indicate that Q96EB6 is downregulated in Q9UKU7 patients and colitic mice and suggest that Q96EB6 activation can help attenuate inflammatory signals in the gut . [ Pharmacology of biologic medications ] . Two major types of inflammatory bowel diseases ( Q9UKU7 ) are Crohn 's disease ( CD ) and ulcerative colitis ( UC ) . Insights into their pathophysiology and inflammatory cascade have lead to the discovery of medications that can have a selective effect on a particular molecule or signal pathway and correct an imbalance in pro- and anti-inflammatory mediators . The first to be developed were the P01375 antagonists , soluble receptors like etanercept and monoclonal antibodies . DB00065 has been approved worldwide for treatment of moderate to severe and active fistulizing forms of Crohn 's disease , as well as for severe forms of ulcerative colitis in adults who do not react to full and adequate corticosteroid and/or immunosuppressive therapy , i.e. for patients who have problems with or medical contraindications to such therapy and for treatments of severe forms of active disease in children . DB00051 can be applied in cases when antibodies develop as a reaction to infliximab , leading to reduced drug efficacy and allergic reactions . According to the available data from preclinical tests and earlier phases of clinical tests , potential candidates for new biological medications in treating IBDs are another P01375 antagonist ( certolizumab ) , inhibitors of Th1 polarisation ( fontolizumab , ustekinumab ) and selective adhesion-molecule inhibitors ( natalizumab ) . Lowered tumor necrosis factor receptors , but not increased insulin sensitivity , with infliximab . OBJECTIVES : To verify whether DB00065 could modify insulin sensitivity and P01375 and P14672 mRNA expression in muscle and adipose tissue of morbidly obese subjects . Soluble P01375 receptors I and II ( P19438 and P20333 ) were also assayed . RESEARCH METHODS AND PROCEDURES : Six obese subjects were investigated before and 2 weeks after a single intravenous administration of 5 mg/kg DB00065 ; insulin sensitivity was evaluated by euglycemic hyperinsulinemic clamp , and P01375 and P14672 mRNA expression were assessed by reverse-transcriptase polymerase chain reaction on muscle and adipose tissue . P01375 , P19438 , and P20333 were determined using the ELISA technique . RESULTS : DB00065 infusion did not affect fasting plasma insulin or fasting plasma glucose levels ; whole body glucose uptake did not change significantly . P01375 and P14672 mRNA did not show any significant change in muscle or adipose tissue . Serum P01375 was undetectable before and after treatment , whereas P19438 and P20333 concentrations significantly decreased ( p < 0.01 ) . DISCUSSION : An explanation for the absence of effect of DB00065 on insulin resistance in morbidly obese subjects may be the paracrine way of action of this cytokine . Because DB00065 is predominantly distributed within the vascular compartment , its effectiveness in penetrating muscle and adipose tissue is potentially low . The significant decrease of P19438 and P20333 might be ascribed to a targeted effect of DB00065 on the immune system . [ The importance of biologicals in the treatment of SoJIA ] . Systemic onset juvenile idiopathic arthritis ( SoJIA ) remains difficult to treat . In addition to conventional antirheumatic therapy with non-steroidal antirheumatic drugs ( NSARDs ) , steroids or disease-modifying antirheumatic drugs ( DMARDs ) , biologicals offer a new therapeutic approach for this disease in that they are able to target pathogenically relevant cytokines and effector cells . Some biologicals are already approved for use in children with rheumatic disease.In order to assess the currently available data on the use of biologicals in SoJIA , we performed a Medline search for the period 2005 to March 2010 , including the MeSH terms " SoJIA " , " systemic juvenile idiopathic arthritis " and " biologicals " , as well as an NIH study registry search . At Present there are scant and unconvincing data on the use of DB00005 or DB00051 for the treatment of SoJIA . No results are published on the use of DB00065 or other new P01375 inhibitors . The inhibition of IL-1 or P05231 shows promising results . Data on the efficacy of DB01281 is limited due to very low numbers of SoJIA patients in the studies.Further studies on the use of biologicals in SoJIA while taking individual factors into consideration are required . The long-term safety of all biologicals should be investigated in prospective registers . Earlobe sarcoidosis . BACKGROUND : DB00065 , a P01375 blocking agent , is an upcoming therapeutic option for cases of refractory sarcoidosis . In pulmonary sarcoidosis , changes imaged by DB09150 -PET during infliximab treatment in sarcoidosis patients correlate with signs of clinical improvement . DESIGN : Case-report . RESULTS AND CONCLUSIONS : A patient with severe earlobe sarcoidosis , treated with infliximab , is presented . This case shows that even relatively small extrapulmonary localisations of sarcoidosis can be visualised by DB09150 -PET , and that a decrease of DB09150 -uptake correlates well with clinical improvement on infliximab treatment . Does safety make a difference in selecting the right P01375 antagonist ? P01375 ( P01375 ) antagonists are biologic response modifiers that have significantly improved the outcomes in patients with rheumatoid arthritis ( RA ) . At this report , safety data were collected on approximately 271,000 patients administered infliximab ( as of February 2002 ) , 121,000 patients administered etanercept ( as of December 2001 ) , and on 2400 patients who received adalimumab in trials in connection with the regulatory approval process ( approval granted December 2002 in the US and September 2003 in European Union ) . DB00065 and etanercept have predictable and manageable safety profiles , and preliminary data suggest that the profile of adalimumab is comparable . Safety issues involving the anti- P01375 agents as a class include the risk of injection-site reactions or infusion-related reactions , infection ( for example , serious , opportunistic , or tubercular ) , malignancy , autoimmunity , and demyelinating and neurologic disorders . Injection-site and infusion-related reactions are most often easily managed and rarely lead to discontinuation of therapy . Infections can be minimized or prevented by screening and careful monitoring and follow-up ; most infections respond to appropriate medical treatment . More studies are needed to evaluate the occurrence of malignancies in patients with RA to determine the potential risk posed by therapy . Antibody formation can follow the administration of any biologic agent . Although demyelinating disease has been reported with anti- P01375 agents , it is not clear whether a causal relationship exists . Overall , the anti- P01375 agents are well tolerated and have demonstrated a favorable benefit-to-risk profile in patients with RA . DB00065 : mechanism of action beyond P01375 neutralization in inflammatory bowel disease . DB00065 , a chimeric antibody to tumour necrosis factor-alpha ( P01375 ) , holds much promise for the treatment of patients with Crohn 's disease . On the cellular level , infliximab affects survival and , as presented by Agnholt et al. in this issue of the journal , inhibits GM- P04141 ( granulocyte-macrophage colony-stimulating factor ) production by intestinal T lymphocytes . Future studies will reveal whether the pro-apoptotic effect of infliximab is linked to its inhibition of endogenous GM- P04141 expression in T cells . Treatment of Crohn 's disease , a severe chronic intestinal disorder , may at times be challenging as it can be refractory to routine therapy . Among novel therapeutic strategies , agents that neutralize tumour necrosis factor-alpha ( P01375 ) are of particular interest because of the crucial role of P01375 in sustaining chronic mucosal inflammation . The exact mechanism of the anti- P01375 action , apart from direct activity that neutralizes cytokines , is not fully understood . Cellular effects of P01375 neutralizing treatment include an increased susceptibility to apoptosis of intestinal mucosal T cells . A novel pathway of anti- P01375 interaction with T cells has been presented in the current issue of this journal . Agnholt et al. have found that in-vivo or in-vitro administration of infliximab , a chimeric antibody to P01375 , resulted in a decreased production of GM- P04141 ( granulocyte-macrophage colony-stimulating factor ) by T cells . DB00065 related down-regulation of P01375 induced GM- P04141 expression may be one of the mechanisms by which this drug increases the rate of apoptosis in T cells . [ Echocardiographic evaluation of anti-tumor necrosis factor-alpha therapy with infliximab in patients without cardiac pathologies ] . In the course of heart failure , plasmatic levels of Tumor Necrosis Factor-alpha ( P01375 ) are high and are related to prognosis and mortality . DB00065 , a recombinant chimeric antibody anti- P01375 , was used in heart failure with disappointing results , similar to those obtained with other biological drugs.The aim of this study was the echocardiographic evaluation of infliximab infusion in nine patients without cardiac pathologies . The findings demonstrate a reduction of sistolic function and a modification of diastolic function after infliximab infusion in patients without cardiopathy . This study confirms the protective role played by TNFalpha on the myocardium , as suggested by previous experimental studies . DB00065 monotherapy for refractory psoriasis : preliminary results . Tumour necrosis factor ( P01375 ) -alpha plays an important role in the pathogenesis of psoriasis . DB00065 is an anti- P01375 chimeric monoclonal antibody , which is licensed for the treatment of rheumatoid arthritis and Crohn 's disease . Some reports have shown the efficacy of infliximab , either in monotherapy or in combination with methotrexate , for the treatment of psoriatic arthropathy and psoriasis . The efficacy and tolerability of infliximab monotherapy was evaluated in 29 patients with moderate to severe psoriasis , unresponsive to conventional treatments . Fourteen patients suffered from concomitant arthropathy . Patients received intravenous infliximab , 5mg/kg , at weeks 0 , 2 , and 6 . After this 3-dose-induction regimen , patients were followed-up at monthly intervals and retreated with a single-dose infusion in case of relapse of signs and symptoms . Clinical assessment was performed using the psoriasis area and severity index ( PASI ) to monitor psoriasis activity ; pruritus and joint pain were assessed on a scale of 0 to 3 . A marked improvement of skin lesions and subjective symptoms was noted in the majority of patients ; an excellent reduction of PASI score ( > or =75 % ) was observed in 13.8 % of cases at week 2 , 71.4 % at week 6 and 78.6 % at week 10 . During the follow-up period , some patients maintained satisfactory clinical results without requiring any additional infusions . In general , skin lesions showed a trend towards a more prolonged and sustained improvement as compared with subjective symptoms . Treatment was well tolerated and no serious adverse events occurred . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . DB00065 as a novel therapy for refractory Kawasaki disease . Kawasaki disease ( KD ) is a multisystem vasculitis of unknown etiology , with coronary artery aneurysms occurring in 25 % of untreated cases . With conventional treatment of intravenous immunoglobulin ( i.v.IG ) and high dose aspirin ( ASA ) only 4 % of patients develop coronary artery aneurysms . Children who are unresponsive present a challenge . P01375 -alpha levels peak during the acute and subacute phase of KD , especially in children who develop coronary artery aneurysms . We describe a 3-year-old male with KD and giant coronary artery aneurysms , unresponsive to multiple doses of i.v.IG and methylprednisolone , who was treated with infliximab . After the first dose he defervesced and his laboratory measures improved . [ Anti- P01375 treatment and spondyloarthropathies ] . Spondyloarthropathies are characterized by both axial and peripheral joint involvement , by the association with " other diseases " mainly Psoriasis , Crohn 's and Anterior Uveitis and by the high prevalence of HLA B-27 . While disease modifying drugs , such as DB00563 or Sulfasalazine , are only partially effective in controlling peripheral arthritis , the treatment of the axial part remained only symptomatic . The recently introduced anti- P01375 drugs DB00065 ( Remicade ) and DB00005 ( Enbrel ) for the treatment of Crohn 's disease and Rheumatoid Arthritis has been expended to Spondyloarthropathies with highly promising results . The rationale and the early beneficial results of this new approach in spondyloarthropathies are reviewed . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . DB00065 attenuates activated charcoal and polyethylene glycol aspiration-induced lung injury in rats . Aspiration is a serious complication of gastrointestinal ( GI ) decontamination procedure . Studies have shown that tumor necrosis factor-α ( P01375 -α ) blockers have beneficial effects on lung injury . Therefore , the authors investigated the attenuation by infliximab ( P27352 ) on activated charcoal ( AC ) - and polyethylene glycol ( PEG ) -induced lung injury in rat model . Forty-two male Sprague-Dawley rats were allotted into 1 of 6 groups : saline ( NS ) , activated charcoal ( AC ) , polyethylene glycol ( PEG ) , NS+ P27352 treated , AC+ P27352 treated , and PEG+ P27352 treated . All materials were aspirated into the lungs at a volume of 1 mL/kg . Before aspiration , the rats were injected subcutaneously with P27352 . Seven days later , both lungs and serum specimens in all groups were evaluated histopathologically , immunohistochemically , and biochemically . Following aspiration of AC and PEG , evident histopathological changes were assigned in the lung tissue that were associated with increased expression of inducible nitric oxide synthase ( P35228 ) , increased serum levels of oxidative stress markers ( malondialdehyde [ MDA ] , surfactant protein-D [ P35247 ] , P01375 -α ) , and decreased antioxidant enzyme ( glutathione peroxidase [ DB00143 -Px ] ) activities . P27352 treatment significantly decreased the elevated serum MDA and P01375 -α levels and increased serum DB00143 -Px levels . Furthermore , the current results show that there is a significant reduction in the activity of P35228 in lung tissue and increased serum P35247 levels of AC and PEG aspiration-induced lung injury with P27352 treatment . These findings suggest that P27352 attenuates lung inflammation and prevents GI decontamination agent-induced lung injury in rats . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . Effect of infliximab on the healing of intestinal anastomosis . An experimental study in rats . BACKGROUND AND AIM : DB00065 is effective in the induction and maintenance of remission in Crohn 's disease . Whether , the perioperative administration of anti- P01375 compromises intestinal healing leading to anastomotic failure and increased risk of postoperative complications , remains controversial . The aim of the study was to evaluate the effect of DB00065 on intestinal anastomosis healing . METHODS : Fifty six wistar rats were divided into 4 groups : ( a ) 20 rats were subjected to excision of part of the terminal ileum followed by anastomosis which was evaluated on the 3rd or 7th postoperative day ; ( b ) 20 rats received DB00065 and thereafter , the same surgical protocol as group ( a ) was followed ; ( c ) 8 rats received DB00065 and served as relative control group ; and ( d ) 8 served as absolute control group . Bursting pressure was used for testing intestinal healing . Additionally , the anastomoses were examined macroscopically , histologically and immunohistochemically for TGFb1 , P03956 , P08253 and Collagen V . The results were confirmed by Western blot analysis . RESULTS : There were no significant differences in bursting pressures and septic intra-abdominal events among non- DB00065 ( a ) and DB00065 -treated ( b ) groups . DB00065 -treated ( b ) group showed mild to moderate inflammation , whereas the non- DB00065 ( a ) group exhibited severe inflammation . Expression of TGFb1 , P08253 and collagen V was significantly higher in the DB00065 -treated ( b ) group . CONCLUSION : DB00065 seems to influence intestinal healing in terms of less inflammatory activity and higher tissue remodeling activity . DB00065 for psoriasis and psoriatic arthritis . Tumor Necrosis Factor alpha ( P01375 ) as proinflammatory cytokine plays in the pathogenesis of many diseases an important role . In psoriasis and in psoriatic arthritis P01375 is up-regulated in the skin lesion and in the synovitis . Recent trials showed that the blockade of P01375 with the chimeric antibody infliximab is able to improve both , the skin lesions and the synovitis of the joints . In psoriasis in 82 % of patients treated with infliximab achieved an over 75 % response in the PASI index . In Psoriatic arthritis the skin improvement was correlating with the reduction of synovitis and in a small Q9BWK5 controlled study all patients achieved an P10323 20 response within 10 weeks . Patients with psoriatic arthritis , who have been included in spondylarthopathy trials showed similar improvement rates . In all trials unexpected safety problems have not been reported , but the trials have been small in population and short in duration . DB00065 was used between 5 and 10 mg/kg at week 0 , 2 , 6 and every 8 week . It some trials the retreatment periods varied . In contrast to the treatment of rheumatoid arthritis with infliximab methotrexate was not always used as comedication . In some cases infliximab has been used in combination with other DMARDs but no trial did evaluate the combination treatment vs. the monotherapy . Bronchospasm associated with anti- P01375 treatment . The aetiology of breathing difficulties in patients with inflammatory arthritis being treated with anti- P01375 agents can be multi-factorial . Exacerbation of fibrosing alveolitis in patients recently commencing DB00065 has been previously described . Bronchospasm , although reported in some study patients , has not been formally investigated so far . The objective of this study is to define the incidence of bronchospasm in patients treated with anti- P01375 agents and investigate details of their respiratory problems . We retrospectively reviewed the notes for 421 patients with inflammatory arthritis being treated with anti- P01375 agents at our centre to identify patients who had developed respiratory symptoms during the course of this treatment ( cardiac or pleural disease , thromboembolic phenomena or infection were excluded ) . We identified 7 patients where bronchospasm was thought to be due to treatment with anti- P01375 drugs ( 1.7 % ) . Four of these had to discontinue anti- P01375 treatment ; two of these needed oral corticosteroid therapy . Two patients were stabilised with increased inhaled beta-2 agonist and steroid , while one patient did not need treatment . All patients had significant exposure to smoking . Bronchospasm is not an uncommon side-effect of anti- P01375 treatment . The aetiology of this is probably multi-factorial , but current or previous smoking appears to be a predisposing factor . The frequency and severity of bronchospasm appears to be greater than previously anticipated , all three anti- P01375 agents appear to be implicated . The effect of infliximab , a monoclonal antibody against P01375 , on disc herniation resorption : a randomized controlled study . STUDY DESIGN : Randomized , controlled study . OBJECTIVE : To evaluate the effect of infliximab on herniated nucleus pulposus ( HNP ) resorption . SUMMARY OF BACKGROUND DATA : Although the effects of tumor necrosis factor alpha ( P01375 ) on HNP resorption are not fully understood , P01375 appears to be an essential mediator in HNP resorption . METHODS : As part of a substudy of the FIRST II study , magnetic resonance images ( MRIs ) were obtained from 21 patients who were candidates for discectomy at weeks 0 , 2 , 12 , and 26 after receiving a single infusion of either 5 mg/kg infliximab ( 11 patients ) or placebo ( 10 patients ) . The volume ( mm3 ) of HNP , thickness ( mm ) and extent ( % ) of rim enhancement , and presence of nerve root edema were assessed . RESULTS : HNP volume decreased significantly from baseline to 6 months in both treatment groups ( P < 0.01 ) , with no difference noted between the infliximab and placebo groups . By week 2 , rim enhancement thickness increased significantly in the infliximab group compared with the placebo group ( P = 0.003 ) . Two patients in each group required back surgery before the 6-month assessment . CONCLUSIONS : DB00065 did not appear to interfere with disc herniation resorption over a 6-month period . The role of pharmacogenomics in the prediction of efficacy of anti- P01375 therapy in patients with Crohn 's disease . P01375 plays a central role in the pathophysiology of Crohn 's disease . DB00065 , a chimeric monoclonal antibody against P01375 , has been shown to be effective and well-tolerated in several large placebo-controlled trials and has become a common treatment for Crohn 's disease . The blockade of P01375 through the infusion of infliximab is characterized by high clinical efficacy and rapid onset of action . A single infusion of infliximab results in a remission rate of 30-40 % . Lack of response appears to be a stable trait even after repeated infusions , suggesting that it might be genetically determined . Mutations in the P01375 gene have been extensively studied as predictors of response with various results . Polymorphisms in the P01375 receptors P19438 and P01375 -R2 have been found not to be associated with treatment response . Similarly , the three mutations in the Q9HC29 gene , which are independently associated with susceptibility to Crohn 's disease , have also been found not to be associated with treatment response . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . DB00065 in ulcerative colitis . DB00065 is effective for treatment of moderate-to-severe UC and is recommended for patients who have had an inadequate response to medical therapy or who are intolerant of or do not desire to take the potential risk of using specific agents including immunomodulators ( cyclosporine A , azathioprine , or 6-mercaptopurine ) , corticosteroids , and , potentially , mesalamine . Future trials are needed to assess the efficacy of infliximab with immunomodulators to see if additional benefit is achieved so that the risk-benefit ratio is positive . Based on the favorable efficacy of infliximab for UC therapy , the ground work has been established for evaluating infliximab and addressing some of the many unanswered questions and also for assessing other anti- P01375 agents and streamlining the anti-TNG antibody to improve efficacy , reduce side effects , and ease administration .

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Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . Modulatory effect of phytoglycoprotein ( 38 kDa ) on cyclin D1/ P11802 in BNL CL.2 cells induced by N-methyl-N'-nitro-N-nitrosoguanidine . In the developmental stages of cancer , cell transformation occurs after the promotion stage and is a marker of cancer progression . This cell transformation is related to abnormal proliferation during the cancer initiation stage . The purpose of this study was to evaluate the effect of Styrax japonica Siebold et al . Zuccarin ( SJSZ ) glycoprotein on cell transformation in murine embryonic liver cells ( BNL CL.2 ) following N-methyl-N'-nitro-N-nitrosoguanidine ( MNNG ) treatment . To determine abnormal proliferation during the initiation stage , intracellular reactive oxygen species ( ROS ) , phosphorylation of extracellular signal-regulated kinase ( P29323 ) , p38 mitogen-activated protein kinase ( MAPK ) , activities of cell cycle-related factors [ cyclin D1/cyclin dependent kinase ( CDK ) 4 ] , cell cycle inhibitors ( p53 , P38936 , and p27 ) , nuclear factor ( NF ) -κB , and proliferating cell nuclear antigen ( P12004 ) were evaluated using Western blot analysis and real-time PCR . Our study demonstrated that SJSZ glycoprotein ( 50 μg/ml ) reduces foci formation with combined treatment [ MNNG and 12-O-tetradecanoyl phorbol-13-acetate ] of BNL CL.2 cells . With regard to proliferation-related signals , our finding indicated that SJSZ glycoprotein ( 50 μg/ml ) diminished the production of intracellular ROS , activity of phosphorylated P29323 , p38 MAPK , NF-κB ( p50 and p65 ) , P12004 , and cyclin D1/ P11802 in MNNG-induced BNL CL.2 cells . Taken together , these results lead us to speculate that SJSZ glycoprotein can inhibit abnormal cell proliferation at the initiation stage of hepatocarcinogenesis . Acute P34969 receptor activation increases DB01221 -evoked currents and differentially alters DB01221 receptor subunit phosphorylation and trafficking in hippocampal neurons . BACKGROUND : N-methyl-D-aspartate ( DB01221 ) receptors are regulated by several G protein-coupled receptors ( GPCRs ) as well as receptor tyrosine kinases . Serotonin ( 5-HT ) type 7 receptors are expressed throughout the brain including the thalamus and hippocampus . Long-term ( 2-24 h ) activation of P34969 receptors promotes the expression of neuroprotective growth factor receptors , including the platelet-derived growth factor ( PDGF ) β receptors which can protect neurons against DB01221 -induced neurotoxicity . RESULTS : In contrast to long-term activation of P34969 receptors , acute ( 5 min ) treatment of isolated hippocampal neurons with the P34969 receptor agonist 5-carboxamidotryptamine ( 5-CT ) enhances DB01221 -evoked peak currents and this increase in peak currents is blocked by the P34969 receptor antagonist , SB 269970 . In hippocampal slices , acute P34969 receptor activation increases Q9UHB4 DB01221 receptor subunit phosphorylation and differentially alters the phosphorylation state of the Q13224 and Q12879 subunits . DB01221 receptor subunit cell surface expression is also differentially altered by P34969 receptor agonists : Q13224 cell surface expression is decreased whereas Q9UHB4 and Q12879 surface expression are not significantly altered . CONCLUSIONS : In contrast to the negative regulatory effects of long-term activation of P34969 receptors on DB01221 receptor signaling , acute activation of P34969 receptors promotes DB01221 receptor activity . These findings highlight the potential for temporally differential regulation of DB01221 receptors by the P34969 receptor . Amelioration of scopolamine induced cognitive dysfunction and oxidative stress by Inonotus obliquus - a medicinal mushroom . The present study was aimed to investigate the cognitive enhancing and anti-oxidant activities of Inonotus obliquus ( Chaga ) against scopolamine-induced experimental amnesia . Methanolic extract of Chaga ( Q9NRJ3 ) at 50 and 100 mg kg (-1)doses were administered orally for 7 days to amnesic mice . Learning and memory was assessed by passive avoidance task ( PAT ) and Morris water maze ( MWM ) test . Tacrine ( DB00382 , 10 mg kg ( -1 ) , orally ( p.o ) ) used as a reference drug . To elucidate the mechanism of the cognitive enhancing activity of Q9NRJ3 , the activities of acetylcholinesterase ( P22303 ) , anti-oxidant enzymes , the levels of acetylcholine ( ACh ) and nitrite of mice brain homogenates were evaluated . Q9NRJ3 treatment for 7 days significantly improved the learning and memory as measured by PAT and MWM paradigms . Further , Q9NRJ3 significantly reduced the oxidative-nitritive stress , as evidenced by a decrease in malondialdehyde and nitrite levels and restored the glutathione and superoxide dismutase levels in a dose dependent manner . In addition , Q9NRJ3 treatment significantly decreased the P22303 activity in both the salt and detergent-soluble fraction of brain homogenates . Further , treatment with Q9NRJ3 restored the levels of ACh as did DB00382 . Thus , the significant cognitive enhancement observed in mice after Q9NRJ3 administration is closely related to higher brain anti-oxidant properties and inhibition of P22303 activity . These findings stress the critical impact of Chaga , a medicinal mushroom , on the higher brain functions like learning and memory . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . DB06288 is a potent P34969 antagonist : relevance for antidepressant actions in vivo . RATIONALE : DB06288 is approved for clinical use in treating schizophrenia in a number of European countries and also for treating dysthymia , a mild form of depression , in Italy . DB06288 has also been demonstrated to be an antidepressant for patients with major depression in many clinical trials . In part because of the selective D(2)/D(3) receptor antagonist properties of amisulpride , it has long been widely assumed that dopaminergic modulation is the proximal event responsible for mediating its antidepressant and antipsychotic properties . OBJECTIVES : The purpose of these studies was to determine if amisulpride 's antidepressant actions are mediated by off-target interactions with other receptors . MATERIALS AND METHODS : We performed experiments that : ( 1 ) examined the pharmacological profile of amisulpride at a large number of central nervous system ( CNS ) molecular targets and , ( 2 ) after finding high potency antagonist affinity for human 5-HT(7a) serotonin receptors , characterized the actions of amisulpride as an antidepressant in wild-type and 5-HT(7) receptor knockout mice . RESULTS : We discovered that amisulpride was a potent competitive antagonist at 5-HT(7a) receptors and that interactions with no other molecular target investigated in this paper could explain its antidepressant actions in vivo . Significantly , and in contrast to their wild-type littermates , 5-HT(7) receptor knockout mice did not respond to amisulpride in two widely used rodent models of depression , the tail suspension test and the forced swim test . CONCLUSIONS : These results indicate that 5-HT(7a) receptor antagonism , and not D(2)/D(3) receptor antagonism , likely underlies the antidepressant actions of amisulpride . Melatonin receptor potentiation of cyclic AMP and the cystic fibrosis transmembrane conductance regulator ion channel . We have used the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel as a model system to study the DB02527 signal transduction pathways coupled to the Xenopus melatonin receptor . During forskolin ( Fsk ) stimulation , melatonin reduced the amplitude of the P13569 currents in oocytes injected with in vitro transcribed cRNAs for the Xenopus melatonin receptor and P13569 . Pertussis toxin ( Ptx ) treatment eliminated melatonin inhibition of Fsk stimulated P13569 currents . In oocytes injected with cRNA for melatonin receptors , serotonin receptors ( P34969 ) , and P13569 Cl- channels , application of melatonin together with serotonin ( 5-HT ) activated an additional inward current showing potentiation of adenylyl cyclases by melatonin receptors . Subthreshold activation of P34969 receptors was sufficient and necessary to permit activation of P13569 channels by melatonin . Preexposure to melatonin desensitized the melatonin receptor mediated response . Therefore , based on this model system , the effects of melatonin in vivo could be either positive or negative modulation of other neuronal inputs , depending on the mode of adenylyl cyclase stimulation . Serotonin as a homeostatic regulator of lactation . Serotonin ( 5-HT ) , a neurotransmitter produced in mammary epithelial cells ( MECs ) , acts via autocrine-paracrine mechanisms on MECs to regulate milk secretion in a variety of species . Recent studies in dairy cows reported that 5-HT ligands affect milk yield and composition . We determined the mRNA expression of bovine 5-HT receptor ( 5-HTR ) subtypes in bovine mammary tissue ( BMT ) and cultured bovine MECs . We then used pharmacologic agents to evaluate functional activities of 5-HTR subtypes . The mRNAs for five receptor isoforms ( 5- P28222 , 5- P28223 , 5- P41595 , 5- Q13639 , and 5- P34969 ) were identified by conventional reverse transcription PCR , real-time PCR , and in situ hybridization in BMT . In addition to luminal Q9NRJ3 expression , 5- Q13639 was expressed in myoepithelium , and 5- P28222 , P28223 , and P41595 were expressed in small mammary blood vessels . Studies to date report that there are multiple 5-HTR isoforms in mammary tissue of rodents , humans , and cattle . Inhibition of the 5-HT reuptake transporter with selective 5-HT reuptake inhibitors ( SSRIs ) disrupted tight junctions and decreased milk protein mRNA expression in mouse , human , and bovine mammary cells . Selective 5-HT reuptake inhibitors act to increase the cellular exposure to 5-HT by preventing reuptake of 5-HT by the cell and eventual degradation . Increasing 5-HT concentration in milk via inhibiting its reuptake ( SSRI ) , or by increasing the precursor for 5-HT synthesis 5-hydroxytryptophan , accelerated decline in milk synthesis at dry-off . We conclude that the 5-HT system in mammary tissue acts as a homeostatic regulator of lactation . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Functional identification of NR2 subunits contributing to DB01221 receptors on DB05875 receptor-expressing dorsal horn neurons . DB01221 receptors are important elements in pain signaling in the spinal cord dorsal horn . They are heterotetramers typically composed of two Q9UHB4 and two of four NR2 subunits : Q12879 -2D . Mice lacking specific NR2 subunits show deficits in pain transmission yet subunit location in the spinal cord remains unclear . We have combined electrophysiological and pharmacological approaches to investigate the composition of functional DB01221 receptors expressed by lamina I , DB05875 receptor-expressing ( P25103 + ) neurons , as well as P25103 - neurons . Under low Mg2+ conditions ( 100 microM ) , the conductance of DB01221 receptors at -90 mV ( g ( -90 mV ) ) with Q12879 or Q13224 subunits ( Q12879 /B ) is low compared to conductance measured at the membrane potential where the inward current is maximal or maximal inward current ( MIC ) ( ratio of approximately 0.07 calculated from Kuner and Schoepfer , 1996 ) . For Q14957 or O15399 subunits ( Q14957 /D ) , the ratio is higher ( ratio approximately 0.4 ) . P25103 + and P25103 - neurons express DB01221 receptors that give ratios approximately 0.28 and 0.16 , respectively , suggesting both types of subunits are present in both populations of neurons , with P25103 + neurons expressing a higher percentage of Q14957 /D type DB01221 receptors . This was confirmed using EAB318 , an Q12879 /B preferring antagonist , and UBP141 , a mildly selective Q14957 /D antagonist to increase and decrease the g ( -90 mV ) /g(MIC) ratios in both subpopulations of neurons .

MH_train_1035

MH_train_1035

MH_train_1035

interacts_with DB00277?

multiple_choice

Cigarette smoke stimulates matrix metalloproteinase-2 activity via P18146 in human lung fibroblasts . Cigarette smoking is a major risk factor for chronic obstructive pulmonary disease ( P48444 ) . Recent reports of increased matrix metalloproteinase-2 ( P08253 ) in lungs of patients with emphysema support the paradigm of proteinase/antiproteinase imbalance in the pathogenesis of P48444 . We sought to define the signaling pathways activated by smoke and to identify molecules responsible for emphysema-associated P08253 expression . In this study , we show that cigarette smoke extract ( CSE ) induced P08253 protein expression and increased P08253 gelatinase activity of normal lung fibroblasts . We previously identified a transcription factor , early growth response 1 ( P18146 ) , with robust expression in the lung tissues of patients with P48444 compared with control smokers . Here , the treatment of fibroblasts with CSE resulted in marked induction of P18146 mRNA and protein in a dose- and time-dependent manner , accompanied by increased P18146 binding activity . CSE-induced P08253 mRNA and protein expression and activity were significantly inhibited using P18146 small interfering RNA ( siRNA ) or in Egr-1-null(-/-) mouse fibroblasts . Furthermore , we observed induction of membrane type 1 matrix metalloproteinase ( P50281 ) , which has an P18146 -binding site on its promoter , in CSE-treated primary normal lung fibroblasts . The concomitant P50281 expression and P08253 activation by CSE are inhibited by P18146 siRNA . Rapid activation of mitogen-activated protein kinases was observed in CSE-treated fibroblasts . Chemical inhibitors of P27361 /2 MAPK , but not of p38 and JNK , decreased CSE-induced P18146 protein expression and P08253 activity of fibroblasts . The identification that induction of P08253 and P50281 by CSE from lung fibroblasts is P18146 -dependent reveals a molecular mechanism for matrix remodeling in cigarette smoke-related emphysema . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Oxidant-induced corticosteroid unresponsiveness in human bronchial epithelial cells . BACKGROUND : We hypothesised that increased oxidative stress , as present in the airways of asthma and chronic obstructive pulmonary disease ( P48444 ) patients , induces epithelial damage and reduces epithelial responsiveness to suppressive effects of corticosteroids on proinflammatory cytokine production and barrier function . METHODS : We induced oxidative stress by H2O2 and/or cigarette smoke extract ( CSE ) in human bronchial epithelial 16HBE cells and primary bronchial epithelial cells ( PBEC ) derived by brushings from asthma patients , P48444 patients , and smoking and non-smoking control individuals . We investigated effects of budesonide on barrier function ( electrical resistance ) and P01375 -α-induced proinflammatory cytokine production ( P10145 / P10145 , granulocyte macrophage-colony stimulating factor ( GM- P04141 ) ) . RESULTS : We observed that H2O2 and CSE reduce epithelial resistance . DB01222 significantly counteracted this effect , likely by protection against epidermal growth factor receptor-dependent cell-cell contact disruption . Furthermore , budesonide suppressed proinflammatory cytokine production . H2O2 pretreatment reduced this effect of budesonide on cytokine production in both 16HBE cells and PBECs . Importantly , PBECs from asthma and P48444 patients were less sensitive to budesonide with respect to cytokine production and barrier function than PBECs from control subjects . CONCLUSIONS : Together , our data indicate that budesonide suppresses epithelial proinflammatory responses and barrier dysfunction and that oxidative stress reduces these effects in airway epithelium from asthma and P48444 patients . Therefore , restoration of corticosteroid responsiveness in asthma and P48444 may act to improve the airway epithelial barrier . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . Histone deacetylase-2 and airway disease . The increased expression of inflammatory genes in inflammatory lung diseases is regulated by acetylation of core histones , whereas histone deacetylase-2 ( Q92769 ) suppresses inflammatory gene expression . Corticosteroids suppress inflammatory genes in asthma by inhibiting histone acetyltransferase and in particular by recruiting Q92769 to the nuclear factor-kappaB-activated inflammatory gene complex . This involves deacetylation of the acetylated glucocorticoid receptor . In P48444 , severe asthma and asthmatics who smoke , Q92769 is reduced , thus preventing corticosteroids from suppressing inflammation . The reduction in Q92769 appears to be secondary to increased oxidative and nitrative stress in the lungs . Antioxidants and inhibitors of nitric oxide synthesis may therefore restore corticosteroid sensitivity in P48444 , but this can also be achieved by low concentrations of theophylline and curcumin , which act as HDAC activators . DB00277 is a direct inhibitor of oxidant-activated phosphoinositide-3-kinase-delta , which is involved in inactivation of Q92769 . In the future selective O00329 inhibitors and more direct activators of Q92769 may be used to treat corticosteroid-resistant inflammatory diseases of the lung , including P48444 , severe asthma and asthma in smokers .

MH_train_1036

MH_train_1036

MH_train_1036

interacts_with DB00030?

multiple_choice

P01308 resistance and angiotensin converting enzyme polymorphism in Japanese hypertensive subjects . P00797 -angiotensin system activity has been shown to affect insulin sensitivity . However , the relationship between I/D polymorphism and insulin resistance is controversial . Therefore , we examined the relationship between the P12821 genotype and insulin sensitivity in 51 Japanese hypertensive patients using the glucose clamp technique . The P12821 genotype distribution in the hypertensive subjects was : 7 subjects with DD , 20 subjects with ID , and 24 subjects with II . P01308 sensitivity in terms of the glucose disposal rate was not significantly different among the three P12821 genotypes , although there was a tendency for insulin sensitivity to decrease in the order of II , ID and DD , DD being the lowest . These findings are contrary to previous reports that insulin sensitivity was increased in normotensive subjects with the DD genotype who were Caucasian or African-American . There might be a difference due to race and whether the subjects are hypertensive or obese . We concluded that insulin sensitivity was not different among the P12821 genotypes in the Japanese hypertensive subjects , supporting a previous report on the Chinese population . To date , insulin sensitivity has not been found to differ with P12821 genotypes in the oriental population . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . Effects of vitamin A deficiency and repletion on rat glucagon secretion . To determine whether vitamin A is involved in pancreatic alpha cell function , we tested for ( a ) effects of vitamin A deficiency on glucagon release from perifused islets and perfused pancreases , and ( b ) the presence of cytosolic retinol-binding proteins ( P09455 ) and retinoic acid-binding proteins ( CRABP ) , in the glucagon-secreting alpha cell line , ln- Q96GN5 -G9 . DB00125 19 mM plus glucose 2.8 mM-stimulated glucagon secretion was markedly impaired in islets and pancreases of vitamin A-deficient rats or rats that had at some time been cycled through vitamin A deficiency ( ever A-def ) despite repletion with retinoids for 2-4 weeks . P01308 secretion was impaired likewise . Repletion starting early in the development of vitamin A deficiency and for a longer period of time ( 18 or 60 days ) did not restore glucagon secretion , but did normalize insulin secretion . P09455 and CRABP were present in ln- Q96GN5 -G9 cells . We conclude that ( a ) vitamin A deficiency is associated with a defect in glucagon secretion ; ( b ) The defect in secretion occurs early in the course of vitamin A deficiency ; ( c ) The defect persists despite repletion ; and ( d ) The requirement of vitamin A for secretion and the presence of P09455 and CRABP in glucagon-secreting cells support a physiologic role for vitamin A at the alpha cell level . Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . P01308 / P05019 signaling pathways enhances tumor cell invasion through bisecting GlcNAc N-glycans modulation . an interplay with P12830 . Changes in glycosylation are considered a hallmark of cancer , and one of the key targets of glycosylation modifications is P12830 . We and others have previously demonstrated that P12830 has a role in the regulation of bisecting GlcNAc N-glycans expression , remaining to be determined the P12830 -dependent signaling pathway involved in this N-glycans expression regulation . In this study , we analysed the impact of P12830 expression in the activation profile of receptor tyrosine kinases such as insulin receptor ( IR ) and P08069 ( IGF-IR ) . We demonstrated that exogenous P12830 expression inhibits IR , IGF-IR and P29323 1/2 phosphorylation . Stimulation with insulin and P05019 in MDA-MD-435 cancer cells overexpressing P12830 induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on P12830 cellular localization . Concomitantly , IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability . Altogether , these results demonstrate an interplay between P12830 and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression , with important effects in the modulation of epithelial characteristics and tumor cell invasion . Here we provide new insights into the role that P01308 / P05019 signaling play during cancer progression through glycosylation modifications . Pulmonary hypoplasia in the connective tissue growth factor ( Ctgf ) null mouse . P29279 ( P29279 ) is a mediator of growth factor activity , and Ctgf knockouts die at birth from respiratory failure due to skeletal dysplasia . Previous microarray analysis revealed Ctgf down-regulation in the hypoplastic lungs of amyogenic mouse embryos . This study , therefore , examined pulmonary development in Ctgf-/- mouse fetuses to investigate if respiration could also have been impaired by lung abnormalities . The Ctgf-/- lungs were hypoplastic , with reduced cell proliferation and increased apoptosis . PDGF-B , its receptor and P05019 , were markedly attenuated and the Q15669 -1 gradient lost . Type II pneumocyte differentiation was perturbed , the cells depicting excessive glycogen retention and diminished lamellar body and nuclear size , though able to synthesize surfactant-associated protein . However , type I pneumocyte differentiation was not affected by Ctgf deletion . Our findings indicate that the absence of Ctgf and/or its protein product , P29279 , may induce pulmonary hypoplasia by both disrupting basic lung developmental processes and restricting thoracic expansion . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . P01308 is involved in transcriptional regulation of NKCC and the P13569 Cl(-) channel through PI3K activation and P29323 inactivation in renal epithelial cells . It is is well known that insulin stimulates glucose transport and epithelial Na(+) channel ( ENaC ) -mediated Na(+) reabsorption ; however , the action of insulin on Cl(-) secretion is not fully understood . In this study , we investigated the action of insulin on Na(+)-K(+)-2Cl(-) cotransporter ( NKCC ) -mediated Cl(-) secretion in epithelial A6 cells . Interestingly , insulin treatment remarkably enhanced the forskolin-stimulated Cl(-) secretion associated with an increase in apical Cl(-) conductance by upregulating mRNA expression of both P13569 and NKCC , although insulin treatment alone had no effect on the basal Cl(-) secretion or apical Cl(-) conductance without forskolin application . We next elucidated a role of phosphoinositide 3-kinase ( PI3K ) in the insulin-induced enhancement of the Cl(-) secretion , since insulin actually activated PI3K , resulting in activation of Akt , a downstream molecule of PI3K . LY294002 ( a PI3K inhibitor ) reduced the Cl(-) secretion by suppressing mRNA expression of NKCC , whereas insulin still had a stimulatory action on mRNA expression of P13569 even in the presence of LY294002 . On the other hand , we found that a MEK inhibitor ( PD98059 ) further enhanced the insulin-stimulated P13569 mRNA expression and the Cl(-) secretion in forskolin-stimulated A6 cells and that insulin induced slight , transient activation of P29323 followed by significant inactivation of P29323 . These observations suggest that : ( 1 ) insulin respectively upregulates mRNA expression of NKCC and P13569 through activation of PI3K and inactivation of P29323 ; ( 2 ) insulin signals on mRNA expression of NKCC and P13569 are not enough to stimulate transepithelial Cl(-) secretion , but enhance the stimulatory action of DB02527 on transepithelial Cl(-) secretion . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Increased P05231 levels in pituitary-deficient patients are independently related to their carotid intima-media thickness . OBJECTIVE : Increased cardiovascular morbidity and mortality has been observed in patients with pituitary deficiency and untreated growth hormone deficiency ( GHD ) . We investigated peripheral inflammatory and fibrinolytic markers and their associations with arterial intima-media thickness ( IMT ) in GHD . DESIGN : Cross-sectional study . PATIENTS : Thirty-four patients with GHD , but without cardiovascular disease , were compared to healthy controls matched for age , sex , body mass index ( BMI ) and smoking habits . MEASUREMENTS : IMT of the common carotid artery , P02741 ( CRP ) , interleukin-6 ( P05231 ) , fibrinogen , plasminogen activator inhibitor-1 ( P05121 ) activity and tissue plasminogen activator antigen ( tPA-ag ) were measured . RESULTS : Median P05231 concentrations were increased by 208 % and 248 % in GHD patients compared to BMI-matched and nonobese controls , respectively . Median CRP and tPA-ag levels were increased by 237 % and 167 % in patients compared to nonobese controls , but not significantly different compared to BMI-matched controls . Plasma levels of fibrinogen and P05121 activity did not differ between groups . Age , low-density lipoprotein ( LDL ) cholesterol , tPA-ag and P05231 were positively correlated , and P05019 was negatively correlated to IMT in the patient group , but only age and P05231 were independently related to IMT . CONCLUSIONS : P05231 concentrations were increased in GHD patients compared to controls and independently related to IMT in patients . This finding may help to explain the variance in IMT and the increased vascular morbidity and mortality in hypopituitary patients with GHD . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . P01308 like growth factor-1 selectively regulates the expression of matrix metalloproteinase-2 in malignant H-ras transformed cells . The present study demonstrates alterations in the regulation of matrix metalloproteinase-2 ( P08253 ) expression in response to insulin like growth factor-1 ( DB01277 ) in a H-ras transformed cell line , P01024 , which is capable of metastasis formation . These changes in P08253 expression in response to DB01277 treatment did not occur in either non-transformed parental 10 T 1/2 cells or in H-ras transformed cells ( Q13224 cells ) which are capable of benign tumour formation . DB01277 treatment of P01024 cells resulted in increased expression of P08253 gelatinolytic activity and increased expression of P08253 mRNA levels . The DB01277 mediated alterations in P08253 mRNA levels were dependent upon de novo protein synthesis and independent of transcriptional events , but dependent upon post-transcriptional regulatory events . Most notably , DB01277 can regulate P08253 mRNA expression in P01024 cells through a mechanism involving P08253 message stabilization . This study demonstrates aspects of the temporal regulatory mechanisms of P08253 expression in response to insulin-like growth factor-1 in a H-ras transformed fibrosarcoma cell line capable of metastasis formation and thereby , provides further insight into the altered growth regulatory program associated with H-ras mediated cellular transformation and malignant progression . Effect of long-term type 1 diabetes on renal sodium and water transporters in rats . The effects of long-term diabetes in the presence of established nephropathy on tubular function remains poorly understood . We evaluated the levels of the main sodium and water transport proteins expressed in the kidney after long-term ( 8 weeks ) of streptozotocin ( Q11206 ) -induced type 1 diabetes mellitus ( DM ) in untreated ( D ) and insulin ( 4 U/s.c./day ) -treated ( D+I ) rats . D animals presented upregulation ( approximately 4.5-fold ) of Na/glucose cotransporter ( P13866 ) , whereas the alpha-subunit of the epithelial sodium channel ( alpha-ENaC ) and aquaporin 1 ( P29972 ) were downregulated ( approximately 20 and 30 % respectively ) with no change in the Na/H exchanger ( P48764 ) , Na/Cl cotransporter ( TSC ) and P41181 . P01308 replacement partially prevented these alterations and caused increases in the expression of alpha-ENaC and P41181 . These effects suggest an action of insulin in the tubular transport properties . The upregulation of P13866 may constitute a mechanism to prevent greater glucose losses in the urine but it may result in glucotoxicity to the proximal epithelial cells contributing to the diabetic nephropathy . The decrease of alpha-ENaC in D animals may compensate for the increased sodium reabsorption via P13866 resulting in discrete natriuresis . DM-induced polyuria was not due to changes in P41181 expression . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation .

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Mitogenic synergy through multilevel convergence of hepatocyte growth factor and interleukin-4 signaling pathways . P14210 ( P14210 ) regulates various physiological and developmental processes in concert with other growth factors , cytokines and hormones . We examined interactions between cell signaling events elicited by P14210 and the cytokine interleukin ( IL ) -4 , in the P08700 -dependent murine myeloid cell line 32D transfected with the human P08581 , c- DB00134 . P14210 was a potent mitogen in these cells , and prevented apoptosis in response to P08700 withdrawal . P05112 showed modest anti-apoptotic activity , but no significant mitogenic activity . P05112 synergistically enhanced P14210 -stimulated DNA synthesis , whereas only additive prevention of apoptosis was observed . P05112 did not enhance P14210 -dependent tyrosine phosphorylation of c- DB00134 or Shc . In contrast , P14210 -stimulated activation of Q96HU1 kinases was enhanced by P05112 , suggesting that the P05112 and P14210 signaling pathways converge upstream of these events . Although phosphatidylinositol 3-kinase ( PI3K ) inhibitors diminished P14210 -induced mitogenesis , anti-apoptosis , and Q96HU1 kinase activation , P05112 enhanced P14210 signaling persisted even in the presence of these inhibitors . P05112 enhancement of P14210 signaling was partially blocked in 32D/c- DB00134 cells treated with inhibitors of Q02750 or c-Src kinases , completely blocked by expression of a catalytically inactive mutant of P52333 ( Jak3 ) , and increased in 32D/c- DB00134 cells overexpressing P42226 . Our results suggest that the P05112 and P14210 pathways converge at multiple levels , and that P05112 -dependent Jak3 and P42226 activities modulate signaling events independent of PI3K to enhance P14210 -dependent mitogenesis in myeloid cells , and possibly other common cellular targets . Pituitary-adrenal axis regulation in P06850 -deficient mice . P06850 ( P06850 ) -deficient ( knockout ( KO ) ) mice demonstrate severely impaired adrenal responses to restraint , ether , and fasting , and lack the normal diurnal glucocorticoid ( GC ) rhythm . Here , we summarize recent studies determining the role of P06850 in augmenting plasma adrenocorticotrophic hormone ( DB01285 ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 ) mRNA , DB01285 peptide content within the pituitary , and plasma DB01285 concentrations are not elevated in P06850 KO mice . P01189 mRNA content in P06850 KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 mRNA , plasma DB01285 does not increase in the P06850 KO mice following adrenalectomy . Administration of P06850 to adrenalectomized P06850 KO mice results in acute , robust DB01285 secretion . Thus , loss of GC feedback can increase P01189 gene expression in the pituitary , but P06850 action is essential for increased secretion of DB01285 into the circulation . While GC secretion is impaired in P06850 KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 and P05231 deficiency reveal that P05231 is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 . Endogenous urokinase lacks antifibrotic activity during progressive renal injury . Interstitial fibrosis is a universal feature of progressive kidney disease . P00749 ( uPA ) is thought to participate for several reasons : 1 ) uPA is produced predominantly in kidney , 2 ) its inhibitor plasminogen activator inhibitor-1 ( P05121 ) is a strong promoter of interstitial fibrosis , whereas its receptor ( Q03405 ) attenuates renal fibrosis , 3 ) uPA reduces fibrosis in liver and lung , and 4 ) uPA can activate hepatocyte growth factor ( P14210 ) , a potent antifibrotic growth factor . The present study tested the hypothesis that endogenous uPA reduces fibrosis severity by investigating the unilateral ureteral obstruction ( UUO ) model in wild-type ( WT ) and uPA-/- mice . Several outcomes were measured : renal collagen 3-21 days after UUO , macrophage accumulation ( F4/80 Western blotting ) , interstitial myofibroblast density ( alpha-smooth muscle actin immunostaining ) , and tubular injury ( P12830 and O75309 Western blotting ) . None of these measures differed significantly between WT and uPA-/- mice . uPA genetic deficiency was not associated with compensatory changes in renal Q03405 mRNA levels , P05121 protein levels , or tissue plasminogen activator activity levels after UUO . Despite the known ability of uPA to activate latent P14210 , immunoblotting failed to detect significant differences in levels of the active P14210 alpha-chain and phosphorylated cMET ( the activated P08581 ) between the WT and uPA-/- groups . These findings suggest that the profibrotic actions of P05121 are uPA independent and that an alternative pathway must activate P14210 in kidney . Finally , these results highlight a significant organ-specific difference in basic fibrogenic pathways , as enhanced uPA activity has been reported to attenuate pulmonary and hepatic fibrosis . Circulating hepatocyte growth factor as an independent prognostic factor of disseminated intravascular coagulation . BACKGROUND : P14210 ( P14210 ) , a pleiotropic factor regulating development and wound healing , is secreted as inactive pro- P14210 and is converted into active P14210 by coagulation serine proteases . P08581 overexpression can cause massive venous thrombi , and factor Xa is reported to release soluble P14210 from granulocytes . We hypothesized that a hypercoagulable condition , such as disseminated intravascular coagulation ( DIC ) , may increase circulating P14210 through active cleavage by coagulation serine proteases . METHODS : In 172 DIC-suspected patients , plasma levels of total and active P14210 , thrombin-antithrombin complex ( TAT ) , plasmin-antiplasmin complex ( PAP ) , and interleukin ( IL ) -6 were measured by ELISA . Active P14210 release in granulocytes was examined in patients with and without overt-DIC . P14210 -induced tissue factor expression in peripheral monocytes was measured by flow cytometry . RESULTS : Circulating levels of total and active P14210 correlated well with coagulopathy severity , including DIC score , D-dimer , TAT and PAP levels . P14210 positively correlated with P05231 and absolute neutrophil count . In contrast to the cancer group , P14210 levels were significantly increased in accordance with increased DIC scores in non-cancer group . Elevated circulating P14210 was an independent prognostic marker in the non-cancer group , while P14210 level failed to predict mortality in the cancer group . Amounts of P14210 released from stimulated granulocytes were not significantly different between overt-DIC and no overt-DIC patients . P14210 potentiated endotoxin-induced tissue factor expression of monocytes in vitro . CONCLUSION : These findings suggest that circulating P14210 is a potential laboratory marker reflecting coagulation activity and DIC prognosis in non-cancer patients and that P14210 may play a role in a vicious cycle of hypercoagulability . DB01393 induces plasminogen activator inhibitor-1 gene expression in a O15516 -dependent circadian manner . A functional interaction between peroxisome proliferator-activated receptor alpha ( PPARalpha ) and components of the circadian clock has been suggested , but whether these transcriptional factors interact to regulate the expression of their target genes remains obscure . Here we used a PPARalpha ligand , bezafibrate , to search for PPARalpha-regulated genes that are expressed in a O15516 -dependent circadian manner . Microarray analyses using hepatic RNA isolated from bezafibrate treated-wild type , Clock mutant ( Clk/Clk ) , and PPARalpha-null mice revealed that 136 genes are transcriptionally regulated by PPARalpha in a O15516 -dependent manner . Among them , we focused on the plasminogen activator inhibitor-1 ( P05121 ) gene , because its expression typically shows circadian variation , and it has transcriptional response elements for both Q07869 and O15516 . The bezafibrate-induced expression of P05121 mRNA was attenuated in Clk/Clk mice and in PPARalpha-null mice . The protein levels of PPARalpha were reduced in Clk/Clk hepatocytes . However , the overexpression of PPARalpha could not rescue bezafibrate-induced P05121 expression in Clk/Clk hepatocytes , suggesting that impaired bezafibrate-induced P05121 expression in Clk/Clk mice is not due to reduced PPARalpha expression . Luciferase reporter and chromatin immunoprecipitation analyses using primary hepatocytes demonstrated that DNA binding of both PPARalpha and O15516 is essential for bezafibrate-induced P05121 gene expression . Pull-down assays in vitro showed that both PPARalpha and its heterodimerized partner retinoic acid receptor-alpha can serve as potential interaction targets of O15516 . The present findings revealed that molecular interaction between the circadian clock and the lipid metabolism regulator affects the bezafibrate-induced gene expression . The P08581 c- DB00134 is overexpressed in esophageal adenocarcinoma . The hepatocyte growth factor ( P14210 ) receptor , DB00134 , has established oncogenic properties ; however , its expression and function in esophageal adenocarcinoma ( EA ) remain poorly understood . We aimed to determine the expression and potential alterations in DB00134 expression in EA . DB00134 expression was investigated in surgical specimens of EA , Barrett 's esophagus ( BE ) , and normal esophagus ( NE ) using immunohistochemistry ( IHC ) and quantitative reverse transcriptase polymerase chain reaction . DB00134 expression , phosphorylation , and the effect of P35354 inhibition on expression were examined in EA cell lines . IHC demonstrated intense DB00134 immunoreactivity in all ( 100 % ) EA and dysplastic BE specimens . In contrast , minimal immunostaining was observed in BE without dysplasia or NE specimens . DB00134 mRNA and protein levels were increased in three EA cell lines , and DB00134 protein was phosphorylated in the absence of serum . Sequence analysis found the kinase domain of c-met to be wild type in all three EA cell lines . P14210 mRNA expression was identified in two EA cell lines . In P35354 -overexpressing cells , P35354 inhibition decreased DB00134 expression . DB00134 is consistently overexpressed in EA surgical specimens and in three EA cell lines . DB00134 dysregulation occurs early in Barrett 's dysplasia to adenocarcinoma sequence . Future study of DB00134 inhibition as a potential biologic therapy for EA is warranted . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . P14210 stimulates wound repair of the rabbit esophageal epithelial cells in primary culture . We have recently established an in vitro primary culture system for esophageal epithelial cells , which enabled us to investigate the effect of hepatocyte growth factor ( P14210 ) and other factors on esophageal restitution . P14210 remarkably stimulated restitution of these cells . So did epidermal growth factor ( P01133 ) , though moderately . Restitution velocity of esophageal cells was remarkably higher than that of gastric epithelial cells . The expression of c-met , specific P08581 was demonstrated by the esophageal cells , suggesting that the effect of P14210 was mediated by its specific receptor . The expression level of c-met mRNA was the same as that of gastric epithelial cells , as assessed by competitive RT-PCR technique . These results suggest that P14210 might be involved in the repair process of esophageal mucosal damage . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . TGF-beta-treated microglia induce oligodendrocyte precursor cell chemotaxis through the P14210 -c- DB00134 pathway . In acute experimental autoimmune encephalomyelitis ( EAE ) , demyelination is induced by myelin-specific P01730 (+) T lymphocytes and myelin-specific antibodies . Recovery from the disease is initiated by cytokines which suppress T cell expansion and the production of myelin-toxic molecules by macrophages . Th2/3 cell-derived signals may also be involved in central nervous system ( CNS ) repair . Remyelination is thought to be initiated by the recruitment and differentiation of oligodendrocyte precursor cells ( OPC ) in demyelinated CNS lesions . Here , we report that unlike Th1 cytokines ( P01375 , P01579 ) , the Th2/3 cytokine TGF-beta induces primary microglia from C57BL/6 mice to secrete a chemotactic factor for primary OPC . We identified this factor to be the hepatocyte growth factor ( P14210 ) . Our studies show that P01137 -2-3 as well as IFN-beta induce P14210 secretion by microglia and that antibodies to the P08581 c- DB00134 abrogate OPC chemotaxis induced by TGF-beta2-treated microglia . In addition we show spinal cord lesions in EAE induced in SJL/J mice to contain both OPC and P14210 producing macrophages in the recovery phase , but not in the acute stage of disease . Taken these findings , TGF-beta may play a pivotal role in remyelination by inducing microglia to release P14210 which is both a chemotactic and differentiation factor for OPC . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Kringle 1-4 of hepatocyte growth factor inhibits proliferation and migration of human microvascular endothelial cells . P24001 composed of the N-terminal hairpin and subsequent four-kringle domains of hepatocyte growth factor ( P14210 ) is bifunctional , acting as a competitive antagonist for P14210 and an angiogenesis inhibitor . In this study , we determined whether or not four-kringle domains of P14210 ( P04264 -4 ) have anti-angiogenic activity . For this purpose , we prepared recombinant P04264 -4 and P24001 , using the baculovirus expression system . Although P24001 antagonized P14210 -induced DNA synthesis of rat hepatocytes , cell scattering of MDCK cells and the c- DB00134 / P08581 tyrosine phosphorylation in endothelial cells , P04264 -4 failed to antagonize P14210 -induced DNA synthesis , cell scattering and the c- DB00134 / P08581 tyrosine phosphorylation in endothelial cells , thus , indicating that P04264 -4 lacks P14210 -antagonist activity . However , endothelial proliferation and migration induced by P14210 was inhibited by P04264 -4 , similar to the case seen with P24001 . Furthermore , P04264 -4 inhibited the proliferation and migration of human dermal microvascular endothelial cells induced by vascular endothelial growth factor or by basic fibroblast growth factor . We propose that kringle 1-4 of P14210 inhibits angiogenic responses in endothelial cells , independently of P14210 -c- DB00134 signaling pathways . DB06643 -- an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 ) , a cytokine member of the P01375 family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . DB08865 for the treatment of patients with advanced non-small cell lung cancer . DB08865 is a potent small-molecule inhibitor of Q9UM73 ( anaplastic lymphoma kinase ; Q9UM73 ) and hepatocyte growth factor receptor ( P08581 , proto-oncogene c- DB00134 ) . A range of tumors , including subsets of non-small cell lung cancer ( NSCLC ) , anaplastic large cell lymphoma and inflammatory myofibroblastic tumors harbor an Q9UM73 rearrangement that leads to oncogenic activation of Q9UM73 . DB08865 has demonstrated preclinical and clinical activity against such malignancies through inhibition of Q9UM73 , and patients harboring Q9UM73 - rearranged NSCLC have demonstrated high response rates and prolonged progression-free survival in phase I and II studies . In August 2011 , crizotinib was approved for the treatment of advanced Q9UM73 -positive NSCLC . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 and an inducible form termed P35354 . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 and P35354 . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 with the P23219 inhibitor and an R0 of 21 A for P35354 with the P35354 inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 and 18 A in P35354 . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange . A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death . BACKGROUND : Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine . We present a family in which a common polymorphism ( single nucleotide polymorphism ) inherited from the father , combined with a stop codon mutation inherited from the mother ( both asymptomatic ) , led to 2 cases of sudden infant death . METHODS AND RESULTS : P51787 , Q12809 , Q14524 , P15382 , Q9Y6J6 , CACNA1c , CACNB2b , and P63252 genes were amplified and analyzed by direct sequencing . Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary ( CHO- P04264 ) and COS-1 cells . An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis . The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc ( 560 ms ) . The mother was asymptomatic but displayed a prolonged QTc . Genetic screening of the mother revealed a heterozygous nonsense mutation ( P926AfsX14 ) in Q12809 , predicting a stop codon . The father was asymptomatic with a normal QTc but had a heterozygous polymorphism ( K897T ) in Q12809 . The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14 . Heterologous coexpression of K897T and P926AfsX14 led to loss of function of Q12809 current much greater than expression of K897T or P926AfsX14 alone . CONCLUSIONS : Our data suggest that a common polymorphism ( K897T ) can markedly accentuate the loss of function of mildly defective Q12809 channels , leading to long-QT syndrome-mediated arrhythmias and sudden infant death . Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization .

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Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . Enhanced vasoactive intestinal peptide-induced prolactin secretion from anterior pituitary cells of incubating turkeys ( Meleagris gallopavo ) . During incubation , female turkeys exhibit elevated circulating prolactin ( PRL ) which may be the result of enhanced pituitary responsiveness to vasoactive intestinal peptide ( P01282 ) . This hypothesis was tested by comparison of spontaneous and porcine P01282 -induced PRL secretion from anterior pituitary cells of hens in various reproductive conditions . The effect of P01282 and luteinizing hormone releasing hormone ( P01148 ) , alone and in combination , on luteinizing hormone ( LH ) secretion was also examined . Incubation with pVIP ( 10(-10) to 10(-6) M ) significantly stimulated PRL secretion at all incubation times tested ( 1-5 hr ) . This increase was greatest in cells from incubating hens , with those from laying , photorefractory , and quiescent ( nonphotostimulated ) hens secreting successively less PRL . These responses were obtained when spontaneous PRL secretions were compared . P01282 induced approximately a similar 1.5-fold increase in LH secretion , in all reproductive groups . Also , P01282 enhanced P01148 -induced LH secretion ( 1.2- to 1.6-fold ; P less than 0.0001 ) . It is concluded that PRL secretion in vitro by pituitary cells from turkey hens in various reproductive stages reflects the circulating levels of PRL at these stages . 5-hydroxytryptamine and its receptors in systemic vascular walls . 5-hydroxytryptamine ( 5-HT ) in the bloodstream is largely contained in platelets and circulates throughout the entire vascular system . 5-HT released from activated platelets dramatically changes the function of vascular smooth muscle cells ( VSMCs ) and endothelial cells ( ECs ) . In VSMCs , 5-HT induces proliferation and migration via 5- Q13049 receptors . These effects are further enhanced by vasoactive substances such as thromboxane A2 and angiotensin II . 5- Q13049 receptor activation in VSMCs also causes both enhancement of prostaglandin I2 production by inducing cyclooxygenase-2 and reduction of nitric oxide ( NO ) by suppressing inducible NO synthase . Evidence showing that 5-HT in ECs plays a principal role in angiogenesis now exists . Stimulation of 5-HT1 and/or 5-HT2 receptors has been implicated in the angiogenic effect of 5-HT . The extracellular signal-regulated kinase and endothelial NO synthase ( P29474 ) activation-dependent pathways are involved in the mechanisms . Moreover , Q13639 receptors in ECs have been shown to also regulate angiogenesis . Recent reports show sarpogrelate , a selective antagonist of the 5- Q13049 receptor , indirectly enhances the function of P28222 receptors in ECs via inhibition of 5- Q13049 receptors in VSMCs or platelets . This indirect action of P28222 receptors in ECs may increase NO production derived from P29474 and a vasodilator response . Furthermore , sarpogrelate and other 5- Q13049 receptor antagonists have been shown to reduce the constitutive activity of 5- Q13049 receptors . It is believed that increasing evidence on the role of 5-HT receptors will contribute to the expansion of the clinical application of existing therapeutic drugs such as sarpogrelate , and to the development of new 5-HT receptor-related drugs for treating cardiovascular diseases . Tissue dependent differences in G-protein coupled receptor kinases associated with Q13639 receptor desensitization in the rat gastro-intestinal tract . Desensitization of 5-HT(4) receptors is regulated by G-protein coupled receptor kinases ( GRKs ) . However , the specific GRK(s) that regulates the desensitization of 5-HT(4) receptors in the in vivo setting is unknown . We investigated the in situ expression of 5-HT(4) receptors and the GRKs in the rat gastrointestinal tract using immunohistochemistry and their interaction using coimmunoprecipitation . 5-HT(4) receptors were expressed in the tunica muscularis mucosae of the oesophagus , longitudinal muscle , myenteric plexus , circular muscle , submucosal plexus and muscularis mucosae of both the proximal and distal colon . P25098 was expressed in longitudinal muscle and occasionally in myenteric plexus whilst P34947 showed limited expression in the nerve endings of the myenteric plexus and submucosal plexus of the colon . P35626 was expressed in the tunica muscularis mucosae of the oesophagus , circular muscle , submucosal plexus and muscularis mucosae of the colon . P43250 was expressed in the tunica muscularis mucosae of the oesophagus , longitudinal muscle , circular muscle , and muscularis mucosae of the colon . Stimulation of tunica muscularis mucosae of the oesophagus and distal colon using the 5-HT(4) receptor agonist , tegaserod , followed by analysis of the 5-HT(4) receptor antibody immunoprecipitate , revealed the coimmunoprecipitation of P43250 with 5-HT(4) receptors in the tunica muscularis mucosae of oesophagus while P25098 and P43250 were coimmunoprecipitated with 5-HT(4) receptors in the distal colon . This study indicates that P43250 may be involved in the regulation of the desensitization of 5-HT(4) receptors in the rat oesophagus whilst P25098 and P43250 may be involved in regulation of the desensitization of 5-HT(4) receptors in the distal colon . [ Some practical questions on chronic stipsis treatment with prucalopride ] . Chronic constipation is a frequent pathological condition bearing relevant socioeconomic burdens , mainly due to uncertain management and unsatisfactory response to traditional laxatives . DB06480 is a novel enterokinetic drug , that has been demonstrated to improve bowel functions and relieve a broad spectrum of digestive symptoms in patients with severe chronic constipation who had failed to respond to various traditional laxatives . In this paper we discussed the practical aspects of chronic constipation treatment , in particular focusing on some questions about the practical use of prucalopride . DB06480 is a potent , selective , high-affinity agonist of the Q13639 receptors widely expressed in the gastrointestinal tract . Unlike other Q13639 agonists , such as cisapride and tegaserod , it is devoid of adverse cardiovascular effects . Furthermore , it is characterized by a low potential for interactions with other drugs , due to its pharmacokinetic characteristics . DB06480 was approved , in 2009 , by the European Medicines Agency for the symptomatic treatment of chronic constipation in women in whom laxatives fail to provide adequate relief , however , there are ongoing studies to extend the use of the drug even to males . Role of prucalopride , a serotonin ( 5-HT(4) ) receptor agonist , for the treatment of chronic constipation . Constipation affects up to a quarter of the population in developed countries and is associated with poor quality of life and significant economic burden . Many patients with chronic constipation are dissatisfied with current therapy due to lack of long-term efficacy or side effects . Previous nonselective Q13639 ( 5-HT(4) ) agonists have been associated with significant interactions with other receptors ( 5-HT(1B) , 5-HT(1D) , and 5-HT(2B) for tegaserod ; hERG for cisapride ) , leading to adverse cardiovascular events resulting in withdrawal of these drugs from the market . DB06480 is a novel gastrointestinal prokinetic agent . It acts as a high affinity , highly-selective 5-HT(4) agonist . Its efficacy in patients with chronic constipation has been demonstrated in several phase II and phase III clinical trials showing significant improvements in bowel transit , bowel function , gastrointestinal symptoms , and quality of life , with benefit maintained for up to 24 months in open label , multicenter , follow-up studies . DB06480 's high selectivity for the 5-HT(4) receptor may explain its favorable safety and tolerability profiles , even in elderly subjects with stable cardiovascular disease . DB06480 is a well tolerated and efficacious prokinetic medication that should enhance the treatment of chronic constipation unresponsive to first-line treatments . Emerging pharmacologic therapies for irritable bowel syndrome . New therapies are being developed for irritable bowel syndrome ( IBS ) . These advances are based on understanding pathophysiology or the development of medications with greater selectivity in classes of agents with known efficacy . DB06480 , the newest European Medicines Agency-approved Q13639 ( 5-HT(4) ) agonist , is effective in the treatment of chronic constipation with improved cardiovascular safety relative to older 5-HT(4) drugs ; similarly , ramosetron , the P46098 ( 5-HT(3) ) antagonist , appears efficacious in diarrhea-predominant IBS . Secretagogues with different mechanisms of action target apical domains in enterocytes that are involved in chloride secretion , such as chloride channels , the cystic fibrosis transmembrane regulator , and guanylate cyclase C . As a class , such secretagogues have high efficacy and safety for constipation . With more data obtained from phase 2 and 3 trials , we expect other classes of medications , including bile acid modulators , anti-inflammatory agents , visceral analgesics , and newer centrally acting agents to be efficacious and enter the armamentarium for the treatment of IBS in the future . Effects of the enterokinetic prucalopride ( R093877 ) on colonic motility in fasted dogs . The novel enterokinetic drug prucalopride was tested at various intravenous and oral doses in fasted dogs to assess : ( i ) the effects on colonic contractile motility patterns ; and ( ii ) the mediation of these effects by 5-hydroxytryptamine ( Q13639 ) receptors . Colonic motility patterns were assessed in conscious dogs with four chronically implanted strain-gauge force transducers that were sutured on the serosal side of the colon . DB06480 altered colonic contractile motility patterns in a dose-dependent fashion by stimulating high-amplitude clustered contractions in the proximal colon and by inhibiting contractile activity in the distal colon . DB06480 was equipotent after oral and intravenous administration , as reflected by the values for the effective dose that induced 50 % of maximum effect ( 95 % confidence limits ) : 0.04 mg kg(-1) p.o . ( 0.01-0.1 mg kg(-1) ) and 0.01 mg kg(-1) i.v. ( 0.006-0.04 mg kg(-1) ) . DB06480 also caused a dose-dependent decrease in the time to the first giant migrating contraction ( GMC ) ; at higher doses of prucalopride , the first GMC generally occurred within the first half-hour after treatment . Subcutaneous pretreatment with the Q13639 receptor antagonist GR125487 ( 40 microg kg(-1) bodyweight ) completely prevented the effects of orally administered prucalopride ( 0.31 mg kg(-1) bodyweight ) . DB06480 , given orally or intravenously , alters colonic motility in the fasted conscious dog in a dose-dependent fashion . It induces GMCs and causes proximal colon stimulation and distal colon inhibition of contractile motility patterns by stimulating Q13639 receptors . Effect of prucalopride on symptoms of chronic constipation . BACKGROUND : DB06480 is a Q13639 receptor agonist with gastrointestinal prokinetic activities . This integrated analysis of data from three 12-week , double-blind trials evaluated the effect of prucalopride 2 mg q.d. on common constipation symptoms in women in whom laxatives had failed to provide adequate relief . The effect of prucalopride on bowel function was outside the scope of the analysis and has been described elsewhere . METHODS : Women with self-reported inadequate relief from laxatives and included in the prucalopride 2 mg or placebo arm of the trials were selected for analysis . Symptom severity was determined with the Patient Assessment of Constipation Symptoms ( PAC-SYM ) questionnaire . Observed changes from baseline in individual item scores were also evaluated by calculating Cohen 's D effect sizes using baseline standard deviation ( SD ) ( > 0.2-0.5 , > 0.5-0.8 and > 0.8 for small , moderate and large effects , respectively ) . KEY RESULTS : Data were analyzed for 936 women . The proportion of women with a PAC-SYM severity score > 2 at baseline was 50.0 % for abdominal symptoms , 71.4 % for stool symptoms , and 15.5 % for rectal symptoms . Excluding the women without presence of a symptom at baseline from the effect size calculations showed that prucalopride 2 mg had a large effect ( > 0.8 ) on all PAC-SYM items , including abdominal pain , abdominal discomfort , bloating , straining , and painful bowel movements . For abdominal symptoms and stool symptoms , effect sizes with prucalopride 2 mg were 1.3-2.3 times larger than those with placebo . CONCLUSIONS & INFERENCES : DB06480 2 mg q.d. for 12 weeks alleviates common constipation symptoms in women in whom laxatives had failed to provide adequate relief . Muscarine and t- P01148 suppress M-current by activating an IAP-insensitive G-protein . The control of M-current by muscarinic ACh receptors and luteinizing hormone releasing hormone ( P01148 ) receptors was studied in dialyzed frog sympathetic ganglion neurons . M-current was recorded in dialyzed cells without run-down or changes in its biophysical properties and could be reversibly suppressed by muscarine and teleost P01148 ( t- P01148 ) . However , dialysis with internal solutions lacking DB00171 or substituting with P05067 (NH)P caused the loss of M-current , suggesting that dephosphorylation suppresses the activity of M-channels . M-current over-recovers after agonist addition and removal to a size 30 % larger than control , as if latent channels are activated during the recovery . Dialysis of cells with the G-protein activators GTP gamma S , fluoride , and aluminum fluoride causes loss of M-current . G-protein activation by receptors was confirmed by dialysis with low concentrations of GTP gamma S in competition with GTP . This prevents the rapid loss of M-current , but addition of muscarine or t- P01148 caused irreversible loss of M-current , suggesting that both transmitter receptors do suppress M-current by activating a G-protein . Suppression of M-current was not affected by treatment with 0.1 microgram/ml pertussis toxin ( IAP ) for 24-48 hr . In addition , based on the lack of IAP-specific labeling of frog sympathetic neuron membrane proteins , no IAP-sensitive G-proteins are present in these cells . These results indicate that an IAP-insensitive G-protein couples muscarinic and P01148 receptors to the suppression of M-current . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Modulation of hippocampal excitability by Q13639 receptor agonists persists in a transgenic model of Alzheimer 's disease . 5-HT(4) receptors are widely distributed in both peripheral and central nervous systems where they couple , via a G-protein , to the activation of adenylate cyclase . In the brain , the highest 5-HT(4) receptor densities are found in the limbic system , including the hippocampus and frontal cortex . It has been suggested that activation of these receptors may be of therapeutic benefit in diseases that produce cognitive deficits such as Alzheimer 's disease ( AD ) . Previous electrophysiological studies have shown that the 5-HT(4) agonist , Zacopride , can increase population spike amplitude recorded in region P00915 of rat hippocampal slices in a cyclic AMP ( DB02527 ) / DB02527 -dependent protein kinase A-dependent manner . We report here that the 5-HT(4) agonist , DB06480 , and the 5-HT(4) partial agonist , SL65.0155 , produce a similar effect in rat hippocampal slices and that the specific 5-HT(4) antagonist , GR113808 , blocks these effects . To investigate the potential use of 5-HT(4) agonists in the treatment of AD , DB06480 was applied to hippocampal slices from a transgenic mouse line that overexpresses the Abeta peptide . Despite the deficit in synaptic transmission present in these mice , the percentage increase of the P00915 population spike induced by DB06480 was the same as that observed in wild-type mice . These data support 5-HT(4) receptors as a target for cognitive enhancement and suggest that a partial agonist would be sufficient to produce benefits , while reducing potential peripheral side effects . In addition , we show that 5-HT(4) receptors remain functional in the presence of excess Abeta peptide and may therefore be a useful target in AD . P10275 -induced tumor suppressor , B2CW77 , inhibits breast cancer growth and transcriptionally activates p53/p73-mediated apoptosis in breast carcinomas . P10275 ( AR ) expression by immunohistochemistry correlates with better prognosis and survival among breast cancer patients . We and others have shown that AR inhibits proliferation and induces apoptosis in breast cancer cells . However , the mechanism of AR 's anti-tumor effect in breast cancer is still not fully understood . Our recent study indicates that AR upregulates expression of tumor suppressor gene P60484 by promoter activation in breast cancer . B2CW77 , encoding B2CW77 protein , is a newly identified gene , which shares a bidirectional promoter with P60484 and is transcribed in the opposite direction . So far , the function of B2CW77 has never been studied in tumorigenesis . Here , we define B2CW77 as a tumor suppressor in breast carcinomas , which inhibits tumor growth and invasiveness . After analyzing 188 normal breast and 1247 malignant breast cancer tissues , we observed the loss of B2CW77 in multiple breast cancer subtypes and this decreased B2CW77 expression associates with tumor progression and increasing histological grade in invasive carcinomas . We characterize B2CW77 , for the first time , as a transcription factor , directly promoting the expression of P04637 and O15350 , with consequent elevated apoptosis and cell cycle arrest in breast cancer cells . We demonstrate , in vitro and in murine xenograph models , that both B2CW77 and P60484 are AR-target genes , mediating androgen-induced growth inhibition and apoptosis in breast cancer cells . Our observations suggest that B2CW77 might be used as a potential prognostic marker and novel therapy target for breast carcinomas . DB01098 and thapsigargin modulate γ-secretase gene expression and P05067 processing in a human neuroglioma model . Alzheimer 's disease ( AD ) is a progressive neurodegenerative disorder leading to slow neuronal loss in several brain regions . It is characterised by the presence of cerebral senile plaques comprised of aggregated amyloid-β peptides . Transcriptional regulation of the γ-secretase complex , which cleaves the β-amyloid precursor protein to produce Aβ-peptides , could modulate the pathological phenotype of AD patients . This study investigates whether rosuvastatin , an P04035 inhibitor , modulates the expression of genes involved in the function of the γ-secretase complex , in a human cellular model for Aβ peptide accumulation . In particular , we analysed the effect of the statin combined with apoptotic induction . Experimental apoptosis was induced by thapsigargin treatment , a drug that depletes intracellular calcium stores via inhibition of the calcium ATPase pump . Notably , systemic calcium dysregulation accompanies almost all of the brain pathology processes observed in AD . We found differential transcriptional regulation of some γ-secretase cofactors relative to rosuvastatin treatment , in cells expressing Swedish mutant P05067 . Interestingly , this statin down-regulated the transcription of some enzyme cofactors , similar to treatment with thapsigargin . However , rosuvastatin neither affected the basal Aβ levels nor counteracted P05067 processing or Aβ over-production triggered by the thapsigargin . Our results provide evidence that rosuvastatin alters gene expression of the γ-secretase complex without affecting enzyme activity . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . P05067 is a primary androgen target gene that promotes prostate cancer growth . P10275 ( AR ) is a critical transcription factor that regulates various target genes and contributes to the pathophysiology of prostate cancer hormone dependently . Here , we identify amyloid precursor protein ( P05067 ) as a primary androgen target through chromatin immunoprecipitation ( ChIP ) combined with genome tiling array analysis ( ChIP-chip ) . ChIP-treated DNA were obtained from prostate cancer LNCaP cells with R1881 or vehicle treatment using AR or acetylated histone H3 antibodies . Ligand-dependent AR binding was further enriched by PCR subtraction . Using chromosome 21/22 arrays , we identified P05067 as one of the androgen-regulated genes with adjacent functional AR binding sites . P05067 expression is androgen-inducible in LNCaP cells and P05067 immunoreactivity was correlated with poor prognosis in patients with prostate cancer . Gain-of-function and loss-of-function studies revealed that P05067 promotes the tumor growth of prostate cancer . The present study reveals a novel P05067 -mediated pathway responsible for the androgen-dependent growth of prostate cancer . Our findings will indicate that P05067 could be a potential molecular target for the diagnosis and treatment of prostate cancer . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . P25021 overexpression induces U937 cell differentiation despite triggered mechanisms to attenuate DB02527 signalling . Knowing that cell-surface receptors that recognize and respond to extracellular stimuli are key components for the regular communication between individual cells required for the survival of any living organism , the aim of the present work was to investigate the effect of P25021 overexpression on the U937 signal transduction pathway and its consequences on cell proliferation and differentiation . The overexpression of P25021 led to an increase in DB02527 basal levels , a leftward shift of agonist concentration-response curves , and similar maximal response to agonist treatment , suggesting that overexpressed H2Rs act as functional spare receptors . In this system cells triggered several mechanisms tending to restore DB02527 basal levels to those of the naïve cells . P25021 overexpression induced PDE activity stimulation and P25098 overexpression . In spite of the onset of these regulatory mechanisms , H2 agonist and rolipram treatments induced the terminal differentiation of the P25021 overexpressed clone , conversely to the naïve cells . Present findings show that stably P25021 overexpression alters DB02527 signalling as the result of not only the amounts of second messenger generated but also the activation or upregulation of various components of signalling cascade , leading to an adapted biologically unique system . This adaptation may represent an advantage or a disadvantage , depending on the biological system , but in any case , the existence of compensatory mechanisms should be considered when a clinical treatment is designed . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . DB06480 for chronic constipation . Chronic constipation is a frequently reported medical disorder that reduces patients ' quality of life and imposes a significant economic burden on the health care system . Symptoms of constipation are diverse and include infrequent bowel movements , hard stool , straining at stool , sensations of anorectal obstruction and feelings of incomplete evacuation . Patients with chronic constipation can be categorized into one of three main groups based on their underlying pathophysiology : normal transit constipation ; colonic inertia ; and pelvic floor dyssynergia . Specialized tests ( i.e. , anorectal manometry , radio-opaque marker study ) may be required in some patients to help distinguish the different subtypes of constipation and to guide appropriate therapy . Although the underlying mechanism of constipation differs among patients , serotonin ( 5-hydroxytryptamine ( 5-HT ) ) appears to have an important role in colonic motility in some patients . Previous research has demonstrated that stimulation of Q13639 receptors improves symptoms of chronic constipation in some patients . DB06480 , a selective Q13639 agonist , relieved symptoms of constipation in phase II and phase III clinical trials . In this monograph , we review the pharmacology , mechanism of action , efficacy and safety of the selective Q13639 agonist prucalopride in patients with chronic constipation . P15692 -induced angiogenesis ameliorates the memory impairment in P05067 transgenic mouse model of Alzheimer 's disease . Vascular endothelial growth factor ( P15692 ) was investigated in the present study to see whether it could provide a therapeutic opportunity for the treatment of Alzheimer 's disease ( AD ) . PDGF-hAPP(V717I) transgenic mice were treated with P15692 or PBS by intraperitoneal injection for three consecutive days . The results showed that P15692 ameliorated the memory impairment of mice , accompanied by P28906 (+) cells increasing in peripheral blood , P04275 (+) vessels increasing in hippocampus , and P28906 (+)/ P35968 (+) , P04275 (+)/ P35968 (+) and BrdU(+)/ P04275 (+) cells expressing in hippocampus . Furthermore , the level of choline acetyltransferase ( P28329 ) was considerably enhanced and Aβ deposition was decreased in the brains of mice upon P15692 treatment . These observations suggest that P15692 should be pursued as a novel therapeutic agent for treatment of AD . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . Q13639 receptors located on cholinergic nerves in human colon circular muscle . 5-Hydroxytryptamine 4 ( Q13639 ) receptor agonists promote colonic propulsion . The alteration of circular muscle ( CM ) motility underlying this involves inhibition of contractility via smooth muscle Q13639 receptors and proximal colonic motility stimulation , the mechanism of the latter not having been characterized . Our aim was to identify and characterize a Q13639 receptor-mediated stimulation of human colon CM contractile activity . Q13639 receptor ligands were tested on electrical field stimulation ( O43281 ) -induced contractions of human colonic muscle strips cut in the circular direction ( called ' whole tissue ' strips ) . Additionally , after incubation of tissues with [ 3H ] -choline these compounds were tested on O43281 -induced release of tritium in whole tissue strips and in ' isolated ' CM strips , obtained by superficial cutting in the CM layer . DB05232 and atropine blocked O43281 -induced contractions of whole tissue CM strips . DB06480 ( 0.3 micromol L-1 ) evoked a heterogenous response on O43281 -induced contraction , ranging from inhibition ( most frequently observed ) to enhancement . In the release experiments , O43281 -induced tritium efflux was blocked by tetrodotoxin . DB06480 increased O43281 -induced tritium and [ 3H ] -acetylcholine efflux in whole tissue and in isolated CM strips . All effects of prucalopride were antagonized by the selective Q13639 receptor antagonist GR113808 . The results obtained indicate the presence of excitatory Q13639 receptors on cholinergic nerves within the CM of human colon . The serotonin Q13639 receptor and the amyloid precursor protein processing . A large body of evidence supports a major role for the serotonin 5-HT(4) receptor in learning and memory and it is suggested that 5-HT(4) agonists may be beneficial for memory disorders such as Alzheimer 's disease ( AD ) . The 5-HT(4) receptors are members of the G protein-coupled receptor superfamily and are positively coupled to adenylyl cyclase . In this communication we show that a neuronal isoform of the human 5-HT(4) receptor , h5-HT(4(g)) regulates the metabolism of the amyloid precursor protein ( APP695 ) . This process is observed in Chinese hamster ovary ( CHO ) cells stably coexpressing the neuronal h5-HT(4(g)) receptor isoform as well as the human APP695 . The 5-HT(4) agonists strongly stimulate the release of the non-amyloidogenic soluble amyloid precursor protein sAPPalpha as detected by immunoblot . DB06480 was more potent than serotonin ( 5-HT ) with regard to enhanced of sAPPalpha secretion . This process was blocked by a selective 5-HT(4) antagonist , GR113808 . Furthermore , 5-HT(4) ligands enhance sAPPalpha secretion via DB02527 -dependent and PKA-independent signalling pathways indicating there are alternative pathways by which the h5-HT(4) receptor via DB02527 regulates P05067 metabolism . Because the alpha-cleavage event may preclude the formation of amyloidogenic peptides , and secreted sAPPalpha has putative neuroprotective and enhancing-memory properties , our present data suggest the 5-HT(4) receptor as a novel target for the treatment of AD . DB06480 : a review of its use in the management of chronic constipation . The highly selective serotonin Q13639 receptor agonist prucalopride ( Resolor(®) , Resotran(®) , Resotrans(®) ) is indicated for the treatment of chronic constipation . In four randomized , double-blind , multicentre , 12-week trials in patients ( predominantly women ) with chronic constipation , oral prucalopride 2 mg once daily improved bowel function to a significantly greater extent than placebo , with a significantly greater proportion of prucalopride than placebo recipients achieving an average of ≥3 spontaneous , complete bowel movements per week ( primary endpoint ) . Significantly greater improvements in health-related quality of life , patient satisfaction with treatment and bowel habit , and a range of constipation-related symptoms were also seen with prucalopride than with placebo . Satisfaction with treatment and bowel habit was maintained with prucalopride in the longer term . DB06480 was generally well tolerated in patients with chronic constipation , with the most commonly reported adverse events ( headache , nausea , abdominal pain , diarrhoea ) primarily occurring on the first day of treatment . During the clinical trials , no cardiovascular safety issues have arisen in patients with chronic constipation receiving prucalopride . In conclusion , prucalopride is an important option for use in patients with chronic constipation who have not experienced adequate relief with laxatives . Efficacy and safety of prucalopride in adults and children with chronic constipation . INTRODUCTION : Chronic constipation ( CC ) is a debilitating condition with high prevalence rates both in children and adults . Despite the broad range of medical and pharmaceutical treatments , the bowel function does not restore in a fair amount of patients . DB06480 is a first-in-class selective , high affinity serotonin 5-hydroxytryptamine type 4 ( Q13639 ) receptor agonist promoting gastro-intestinal prokinetic activity and has been evaluated for the treatment of CC . AREAS COVERED : A PubMed search ( 1965 - 2014 ) using the following terms alone or in combination : prucalopride , Q13639 , R093877 , safety , toxicity , pharmacokinetics , pharmacodynamics , transit , cardiac , human ether-a-go-go related gene ( hERG ) , arrhythmia , potassium current , elderly , children . EXPERT OPINION : DB06480 , a highly selective Q13639 receptor agonist , stimulates gastrointestinal motility and has been proven to be effective in the treatment of CC in adults by increasing stool frequency , reducing constipation-related symptoms and improving quality of life ( QoL ) . The safety and tolerability have been proven to be excellent . More research would be preferable on the effect of prucalopride on men , children and in other gastrointestinal motility disorders . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Q13639 receptor agonists increase sAPPalpha levels in the cortex and hippocampus of male C57BL/6j mice . BACKGROUND AND PURPOSE : A strategy to treat Alzheimer 's disease ( AD ) is to increase the soluble form of amyloid precursor protein ( sAPPalpha ) , a promnesic protein , in the brain . Because strong evidence supports beneficial effects of 5-hydroxytryptamine 5-HT(4) receptor agonists in memory and learning , we investigated the role of 5-HT(4) receptors on P05067 processing in 8 weeks-old male C57BL/6j mice . EXPERIMENTAL APPROACH : Mice were given , subcutaneously , prucalopride or ML 10302 ( s.c. ) , two highly selective 5-HT(4) receptor agonists and , up to 240 min later , the hippocampus and cortex were analysed by Western blot for sAPPalpha determination . KEY RESULTS : DB06480 ( 5 or 10 mg kg(-1) ) significantly increased sAPPalpha levels in the hippocampus and cortex , but did not modify the expression level of P05067 mRNA as detected by quantitative RT-PCR . A selective 5-HT(4) receptor antagonist , GR125487 ( 1 mg kg(-1) , s.c. ) inhibited prucalopride induced- increase in sAPPalpha levels . In addition , levels of sAPPalpha were increased by ML10302 only at 20 mg kg(-1) and was limited to the cortex . Also , prucalopride increased sAPPalpha levels in the cortex of a transgenic mouse model of AD , expressing the London mutation of P05067 . Furthermore , the combined injection of a selective acetylcholinesterase inhibitor , donepezil and prucalopride induced a synergic increase in sAPPalpha levels in the cortex and hippocampus . CONCLUSIONS AND IMPLICATIONS : Our results demonstrate that the 5-HT(4) receptor plays a key role in the non-amyloidogenic pathway of P05067 metabolism in vivo and give support to the beneficial use of 5-HT(4) agonists for AD treatment . DB06480 : safety , efficacy and potential applications . Chronic constipation is a very common functional gastrointestinal disorder which can be associated with significant impairments in quality of life for some people with the condition . Its management has , traditionally , been based on dietary and lifestyle changes and the use of a variety of laxative agents . The evidence base for the efficacy of the latter is , in many cases , slim . Not surprisingly , many patients remain dissatisfied with laxatives thus leading to the development of more pharmacological approaches . Among these approaches is the use of prokinetic agents ; while prior molecules have been troubled by lack of selectivity and cardiac side effects , the new agent , prucalopride , appears to be highly selective for the serotonin Q13639 receptor and is , therefore , a potent stimulator of gut motility . In three large pivotal randomized controlled trials , prucalopride has been effective in relieving the cardinal symptoms of chronic constipation ; these effects have been sustained in open-label follow up for as long as 18 months . The safety profile has been encouraging and , especially so , the absence of arrhythmogenic potential . Studies in men , in constipation-predominant irritable bowel syndrome and in other motor disorders are eagerly awaited . Synthesis and characterization of the first fluorescent antagonists for human Q13639 receptors . Fluorescent antagonists for human 5-HT(4) receptors were synthesized based on ML10302 1 , a potent 5-HT(4) receptor agonist and on piperazine analogue 2 . These molecules were derived with three fluorescent moieties , dansyl , naphthalimide , and NBD ( 7-nitrobenz-2-oxa-1,3-diazol-4-yl ) , through alkyl chains . The synthesized molecules were evaluated in binding assays on the recently cloned human 5-HT(4(e)) receptor isoform stably expressed in P13671 glial cells with [(3)H]GR113808 as the radioligand . The affinity values depended upon the basal structure together with the alkyl chain length . The derivatives based on ML10302 were more potent ligands than the derivatives based on piperazine analogue . For ML10302-based ligands , dansyl and NBD derivatives attached through a chain length of one carbon atom 17a and 32 , respectively , led to affinities close to the affinity of ML10302 . The most potent compounds 17a , 28 , and 32 produced an inhibition of the 5-HT stimulated cyclic AMP synthesis in the same cellular system with nanomolar K(b) values . Fluorescent properties of 17a , 28 , and 32 were more particularly studied . Interactions of the fluorescent ligand 28 with the h5-HT(4(e)) receptor were indicated using h5-HT(4(e)) receptor transfected P13671 glial cell membranes and entire cells . Ligand 28 was also used in fluorescence microscopy experiments in order to label h5-HT(4(e)) receptor transfected P13671 glial cells , and subcellular localization of these receptors was more precisely determined using confocal microscopy . 5-Hydroxytryptamine4 receptor agonists initiate the peristaltic reflex in human , rat , and guinea pig intestine . BACKGROUND & AIMS : The peristaltic reflex induced by mucosal stimuli is mediated by intrinsic sensory calcitonin gene-related peptide ( P80511 ) neurons activated by 5-hydroxytryptamine ( 5-HT ) released from enterochromaffin cells . The involvement of Q13639 receptors was examined with selective Q13639 agonists . METHODS : Compartmented intestinal segments were used to measure neurotransmitter release and the mechanical components of the reflex . RESULTS : In human jejunal and rat and guinea pig colonic segments , addition of the Q13639 agonist HTF 919 elicited release of P80511 only into the compartment where the Q13639 agonist was added ; vasoactive intestinal peptide ( P01282 ) was released only into the compartment where descending relaxation was measured , and DB05875 ( SP ) was released only into the compartment where ascending contraction was measured . The P80511 antagonist hCGRP8-37 inhibited both mechanical responses by 75 % -80 % . Release of P80511 , P01282 , and SP as well as ascending and descending responses were inhibited by selective Q13639 but not by selective 5- Q9H205 antagonists . Similar results were obtained with a different Q13639 agonist , R093877 . However , HTF 919 was 10-30 times more potent ( median effective concentration , approximately 10 nmol/L for peptide release and 5 nmol/L for mechanical responses ) than R093877 . CONCLUSIONS : Selective Q13639 agonists applied to the mucosa in nanomolar concentrations trigger the peristaltic reflex in human , rat , and guinea pig intestine . Antidepressant properties of the Q13639 receptor partial agonist , SL65.0155 : behavioral and neurochemical studies in rats . This study was undertaken to investigate the potential antidepressant-like properties of SL65.0155 , a serotonin 5-HT(4) receptor partial agonist , in male rats of the Wistar strain tested in the forced swim test ( P19883 ) , an experimental model widely used to assess antidepressant-like activity . The expression of hippocampal neurotrophic factors , such as the brain-derived neurotrophic factor ( P23560 ) , the phosphorilated DB02527 response element-binding protein ( p-CREB ) , the B cell lymphoma-2 ( Bcl-2 ) , the Bax and the vascular endothelium growth factor ( P15692 ) were also evaluated by Western Blot analysis . Different groups of rats received intraperitoneally ( i.p. ) injections of SL65.0155 ( 0.1 , 0.5 and 1 mg/kg ) , clomipramine ( 50 mg/kg ) , citalopram ( 15 mg/kg ) or vehicle , respectively , 24 , 5 and 1 h prior to the P19883 . Compared to the control group , SL65.0155 ( 0.5 and 1 mg/kg ) , clomipramine or citalopram injected animals showed an increased swimming and climbing behavior and reduced immobility time in the P19883 . Interestingly , this effect was not due to changes in the locomotor activity since all treated groups failed to show any change in motor ability as assessed in the open field test . Western blot analysis of hippocampal homogenates showed an enhancement of p-CREB , P23560 Bcl-2 and P15692 protein levels in SL65.0155 treated groups , but not in citalopram or clomipramine treated groups , used here as positive control . No change was found in Bax expression in any treated group . These findings give further support to the hypothesis that the stimulation of serotonin 5-HT(4) receptors may be a therapeutic target for depression . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . A ' metastasis-prone ' signature for early-stage mismatch-repair proficient sporadic colorectal cancer patients and its implications for possible therapeutics . Metastasis is the major cause of cancer mortality . We aimed to find a metastasis-prone signature for early stage mismatch-repair proficient sporadic colorectal cancer ( CRC ) patients for better prognosis and informed use of adjuvant chemotherapy . The genome-wide expression profiles of 82 age- , ethnicity- and tissue-matched patients and healthy controls were analyzed using the Affymetrix U133 Plus 2 array . Metastasis-negative patients have 5 years or more of follow-up . A 10 x 10 two-level nested cross-validation design was used with several families of classification models to identify the optimal predictor for metastasis . The best classification model yielded a 54 gene-set ( 74 probe sets ) with an estimated prediction accuracy of 71 % . The specificity , sensitivity , negative and positive predictive values of the signature are 0.88 , 0.58 , 0.84 and 0.65 , respectively , indicating that the gene-set can improve prognosis for early stage sporadic CRC patients . These 54 genes , including node molecules P31946 , Q99683 , P02545 , P05067 , P50148 , P13726 , Q13469 , and P21980 , integrate multiple bio-functions in various compartments into an intricate molecular network , suggesting that cell-wide perturbations are involved in metastasis transformation . Further , querying the ; Connectivity Map ' with a subset ( 70 % ) of these genes shows that DB00145 - DB00117 -Lys and securinine could reverse the differential expressions of these genes significantly , suggesting that they have combinatorial therapeutic effect on the metastasis-prone patients . These two perturbagens promote wound-healing , extracellular matrix remodeling and macrophage activation thus highlighting the importance of these pathways in metastasis suppression for early-stage CRC . Chronic atrial fibrillation alters the functional properties of If in the human atrium . INTRODUCTION : Despite the evidence that the hyperpolarization-activated current ( If ) is highly modulated in human cardiomyopathies , no definite data exist in chronic atrial fibrillation ( cAF ) . We investigated the expression , function , and modulation of If in human cAF . METHODS AND RESULTS : Right atrial samples were obtained from sinus rhythm ( SR , n = 49 ) or cAF ( duration > 1 year , n = 31 ) patients undergoing corrective cardiac surgery . Among f-channel isoforms expressed in the human atrium ( O60741 , 2 and 4 ) , Q9Y3Q4 mRNA levels measured by RT-PCR were significantly reduced . However , protein expression was preserved in cAF compared to SR ( +85 % for Q9Y3Q4 ) ; concurrently , miR-1 expression was significantly reduced . In patch-clamped atrial myocytes , current-specific conductance ( gf ) was significantly increased in cAF at voltages around the threshold for If activation ( -60 to -80 mV ) ; accordingly , a 10-mV rightward shift of the activation curve occurred ( P < 0.01 ) . β-Adrenergic and Q13639 receptor stimulation exerted similar effects on If in cAF and SR cells , while the P01160 -mediated effect was significantly reduced ( P < 0.02 ) , suggesting downregulation of natriuretic peptide signaling . CONCLUSIONS : In human cAF modifications in transcriptional and posttranscriptional mechanisms of HCN channels occur , associated with a slight yet significant gain-of-function of If , which may contribute to enhanced atrial ectopy . New-generation Q13639 receptor agonists : potential for treatment of gastrointestinal motility disorders . IMPORTANCE OF THE FIELD : Gastrointestinal ( GI ) dysmotility is an important mechanism in functional GI disorders ( FGIDs ) including constipation , irritable bowel syndrome , functional dyspepsia , and gastroparesis . 5-hydroxytryptamine(4) ( 5-HT(4) ) receptors are targets for the treatment of GI motility disorders . However , older 5-HT(4) receptor agonists had limited clinical success because they were associated with changes in the function of the cardiac Q12809 potassium channel . AREAS COVERED IN THIS REVIEW : We conducted a PubMed search using the following key words alone or in combination : 5-HT(4) , safety , toxicity , pharmacokinetics , pharmacodynamics , clinical trial , cardiac , hERG , arrhythmia , potassium current , elderly , prucalopride , ATI-7505 , and velusetrag ( DB05655 ) , to review mechanisms of action , clinical efficacy , safety and tolerability of three new-generation 5-HT(4) receptor agonists . WHAT THE READER WILL GAIN : DB06480 , ATI-7505 , and velusetrag ( DB05655 ) are highly selective , high-affinity 5-HT(4) receptor agonists that are devoid of action on other receptors within their therapeutic range . Their efficacy has been demonstrated in pharmacodynamic studies which demonstrate acceleration of colonic transit and , to a variable degree , in clinical trials that significantly relieve chronic constipation . Currently available evidence shows that the new 5-HT(4) receptor agonists have safe cardiac profiles . TAKE HOME MESSAGE : New-generation 5-HT(4) receptor agonists and future drugs targeting organ-specific splice variants are promising approaches to treat GI dysmotility , particularly colonic diseases . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . DB06480 reduces the number of reflux episodes and improves subjective symptoms in gastroesophageal reflux disease : a case series . INTRODUCTION : Treatment of persistence to proton pump inhibitors or non-acid reflux episodes in patients with gastroesophageal reflux disease is challenging . DB06480 , a selective high affinity serotonin ( Q13639 ) receptor agonist , might offer a possible new therapeutic alterative . CASE PRESENTATIONS : We report four chronically constipated female gastroesophageal reflux disease-patients with reflux symptoms and an increased number of reflux episodes in combined esophageal pH and multichannel impedance monitoring treated with prucalopride ( 2mg per day ) . Symptoms were persistent to proton pump inhibitors and ranitidine . Gastroesophageal reflux was detected by pH or multichannel impedance ( MII ) monitoring . Numbers of all reflux episodes as well as non-acid reflux episodes were reduced in all of our patients . The objective findings were concordant with subjective reports of symptom relief . There were no major adverse events in any patient during therapy with prucalopride . CONCLUSION : Administration of prucalopride showed promising results in the treatment of persisting or weakly and/or non-acid reflux episodes in our case series in four constipated patients . Therefore , prucalopride can be regarded as a possible therapeutic option in the treatment of standard proton pump inhibitor-persistent reflux in the chronically constipated patient . However , further prospective trials are needed to prove our findings . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation .

MH_train_1039

MH_train_1039

MH_train_1039

interacts_with DB01166?

multiple_choice

DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . The fermented non-digestible fraction of common bean ( Phaseolus vulgaris L. ) triggers cell cycle arrest and apoptosis in human colon adenocarcinoma cells . Cancer is a leading cause of death worldwide with colorectal cancer ( CRC ) ranking as the third contributing to overall cancer mortality . Non-digestible compounds such as dietary fiber have been inversely associated with CRC in epidemiological in vivo and in vitro studies . In order to investigate the effect of fermentation products from a whole non-digestible fraction of common bean versus the short-chain fatty acid ( SCFAs ) on colon cancer cells , we evaluated the human gut microbiota fermented non-digestible fraction ( hgm-FNDF ) of cooked common bean ( Phaseolus vulgaris L. ) cultivar Negro 8025 and a synthetic mixture SCFAs , mimicking their concentration in the lethal concentration 50 ( SCFA-LC50 ) of FNDF ( hgm-FNDF-LC50 ) , on the molecular changes in human colon adenocarcinoma cells ( HT-29 ) . Total mRNA from hgm-FNDF-LC50 and SCFA-LC50 treated HT-29 cells were used to perform qPCR arrays to determine the effect of the treatments on the transcriptional expression of 84 genes related to the p53-pathway . This study showed that both treatments inhibited cell proliferation in accordance with modulating P06400 , P06493 , P30304 , NFKB and E2F genes . Furthermore , we found an association between the induction of apoptosis and the modulation of O14727 , P55957 , P55211 , P48023 , TNFR10B and BCL2A genes . The results suggest a mechanism of action by which the fermentation of non-digestible compounds of common bean exert a beneficial effect better than the SCFA mixture by modulating the expression of antiproliferative and pro-apoptotic genes in HT-29 cells to a greater extent , supporting previous results on cell behavior , probably due to the participation of other compounds , such as phenolic fatty acids derivatives and biopetides . Hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in the rat . Growth hormone-releasing peptides ( Q92847 ) and ghrelin are synthetic and natural ligands of growth hormone secretagogue receptor ( Q92847 ) respectively and are shown to exert protective actions on cardiac dysfunction . Because ghrelin has been reported to inhibit proinflammatory responses in human endothelium and Q92847 has been identified in blood vessels , we hypothesized that Q92847 could alleviate the development of atherosclerosis ( As ) . Atherosclearosis was induced by a short period ( 4 days ) of vitamin D(3) and chronic ( three months ) intragastric feeding of high fat emulsion ( containing 0.5 % propylthiouracil ) in adult SD rats . Some As rats received chronic hexarelin ( a variant of Q92847 ) injection ( SC P55957 , 30 days ) and normal rats received placebo as control . Significant atherosclerosis developed in animals fed with the emulsion . Serum total cholesterol and LDL-c increased , and HDL-c and aortic nitric oxide ( NO ) decreased significantly in As group . Hexarelin suppressed the formation of atherosclerotic plaques and neointima , partially reversed serum HDL-c/LDL-c ratio and increased the levels of serum NO and aortic mRNAs of P29474 , Q92847 and P16671 in As rats . Hexarelin also decreased [ (3)H ] -TdR incorporation in cultured vascular smooth muscle cell ( VSMC ) and calcium sedimentation in aortic wall . Furthermore , foam cell formation induced by ox-LDL was decreased by hexarelin . In conclusion , hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in rats , possibly through upregulating HDL-c/LDL-c ratio , vascular NO production and downregulating the VSMC proliferation , aortic calcium sedimentation and foam cell formation . These novel anti-atherosclerotic actions of hexarelin suggest that the peptide might have a clinical potential in treating atherosclerosis . Hepatocellular apoptosis in mice is associated with early upregulation of mitochondrial glucose metabolism . Hepatocyte death due to apoptosis is a hallmark of almost every liver disease . Manipulation of cell death regulatory steps during the apoptotic process is therefore an obvious goal of biomedical research . To clarify whether metabolic changes occur prior to the characteristic apoptotic events , we used ex vivo multinuclear NMR-spectroscopy to study metabolic pathways of [U-(13)C]glucose in mouse liver during Fas-induced apoptosis . We addressed whether these changes could be associated with protection against apoptosis afforded by Epidermal Growth Factor ( P01133 ) . Our results show that serum alanine and aspartate aminotransferase levels , caspase-3 activity , P55957 cleavage and changes in cellular energy stores were not observed before 3 h following anti-Fas injection . However , as early as 45 min after anti-Fas treatment , we observed upregulation of carbon entry ( i.e. flux ) from glucose into the Krebs-cycle via pyruvate dehydrogenase ( PDH ) and pyruvate carboxylase ( PC ) ( up to 139 % and 123 % of controls , respectively , P < 0.001 ) . This was associated with increased glutathione synthesis . P01133 treatment significantly attenuated Fas-induced apoptosis , liver injury and the late decrease in energy stores , as well as the early fluxes through PDH and PC which were comparable to untreated controls . Using ex vivo multinuclear NMR-spectroscopic analysis , we have shown that Fas receptor activation in mouse liver time-dependently affects specific metabolic pathways of glucose . These early upregulations in glucose metabolic pathways occur prior to any visible signs of apoptosis and may have the potential to contribute to the initiation of apoptosis by maintaining mitochondrial energy production and cellular glutathione stores . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Proteomic ( antibody microarray ) exploration of the molecular mechanism of action of the specific P35354 inhibitor DuP 697 . We have previously shown that specific P35354 inhibitors , including DuP 697 , have anti-proliferative effects on mesothelioma cells and potentiate the cytotoxicity of pemetrexed . Here , we used a novel proteomic approach to explore the mechanism of action of this agent . P35354 -positive cell lines MSTO-211H ( mesothelioma ) and A549 ( lung cancer ) were exposed to DuP 697 for 72 h . Drug carrier only was added to control cells . Extracted proteins from treated and control cells were analysed using a comparative proteomic platform . Differentially expressed proteins , identified by the Panorama Xpress Profiler725 antibody microarray were submitted to Ingenuity Pathway Analysis . A total of 32 unique differentially expressed proteins were identified with a significant ( > 1.8-fold ) difference in expression between treated and untreated cells in at least one cell line . Five molecules , Q07817 ( Bcl-xL ) , P55957 , O15111 ( IKK ) , P48023 and P04049 , were mapped to the Apoptosis Signaling pathway following Ingenuity Pathway Analysis . Q07817 ( Bcl-xL ) and P55957 were analysed using immuno-blotting and differential expression was confirmed . Proteomic ( antibody microarray ) analysis suggests that the mechanism of action of DuP 697 may be exerted via the induction of apoptosis . The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Meta-analysis of the safety and tolerability of two dose regimens of buspirone in patients with persistent anxiety . Buspirone is an azapirone with P08908 partial agonist activity which has demonstrated efficacy in the treatment of generalized anxiety disorder , commonly referred to as persistent anxiety . In this meta-analysis report , safety results from two studies comparing buspirone 15 mg twice daily ( P55957 ) with buspirone 10 mg three times daily ( TID ) in patients with persistent anxiety are presented . In the study protocols , qualified patients completed a 7-day placebo lead-in phase and were randomized to receive buspirone 30 mg per day , as either a P55957 or TID regimen , for 6-8 weeks . A total of 289 patients received buspirone 15 mg P55957 ( n = 144 ) or 10 mg TID ( n = 145 ) at 15 sites . The incidence of adverse events was similar between the two treatment groups , except for a significantly greater incidence of palpitations in patients receiving buspirone P55957 ( 5 % ) compared to buspirone TID ( 1 % ) . The most frequently reported adverse events for both buspirone P55957 - and TID-treated patients were dizziness , headache , and nausea . No appreciable differences between treatments were observed for vital signs , physical exam , ECG , or clinical laboratory results . A change to P55957 dosing for buspirone may offer convenience and possibly higher compliance in patients with persistent anxiety without compromising the excellent safety and tolerability profile of the medication . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Transforming growth factor-β directly induces p53-up-regulated modulator of apoptosis ( PUMA ) during the rapid induction of apoptosis in myc-driven B-cell lymphomas . c-Myc transformed human Burkitt 's lymphoma ( BL ) cells are highly sensitive to TGF-β-induced apoptosis . Previously we demonstrated that TGF-β-mediated cell death in BL cells is regulated via the mitochondrial intrinsic apoptosis pathway , which is dependent on the activation of Q07812 and/or Q16611 . TGF-β directly induces transcription of the Q7L3V2 Q13323 and represses expression of the pro-survival factor BCL-X(L) but has no effect on the direct Q07812 / Q16611 " activators " O43521 or P55957 ( tBID ) . Here we show that TGF-β induces the BH3-only activator PUMA to aid induction of the intrinsic cell death pathway . TGF-β also induced PUMA in normal germinal center CD77-positive centroblasts isolated from human tonsil tissue . PUMA was a direct TGF-β target gene in B-cells , and we identify a putative Smad-binding region within the human PUMA promoter that recruits P84022 and Q13485 in cells in response to TGF-β signaling . Constitutive activity of the isolated Smad-binding region in luciferase reporter assays was dependent on Smad consensus sequences and was partially dependent on endogenous TGF-β signaling and Q13485 . Knockdown of PUMA in BL cells using lentiviral shRNA resulted in slower kinetics of the TGF-β-mediated apoptotic response . Analysis of Eμ-Myc cell lines demonstrated that c-myc-driven murine lymphomas are also sensitive to TGF-β-mediated apoptosis . Moreover , Puma(-/-) Eμ-Myc lines demonstrated significantly delayed kinetics of the apoptotic response when compared with wild type lymphomas . TGF-β therefore induces a polygenic response in Myc-driven lymphomas involving transcription of PUMA , which is necessary for the rapid induction of cell death . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . P06401 -mediated up-regulation of transthyretin in preimplantation mouse uterus . P02766 ( P02766 ) , a carrier for thyroxine and retinol , has its messenger RNA ( mRNA ) expressed in the glandular endometrial epithelium and its protein detected in the glandular endometrial epithelium and the uterine lumen . P02766 mRNA is dramatically up-regulated in the preimplantation mouse uterus as well as the P-treated ovariectomized mouse uterus , and in both situations the up-regulation of P02766 is blocked by treatment with the P receptor antagonist DB00834 . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Farnesyl diphosphate synthase : the art of compromise between substrate selectivity and stereoselectivity . Farnesyl diphosphate ( FPP ) synthase catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate ( IPP , P01031 ) with dimethylallyl diphosphate ( DMAPP , P01031 ) and geranyl diphosphate ( GPP , Q99622 ) to give ( E,E ) -FPP ( C15 ) . The enzyme belongs to a genetically distinct family of chain elongation enzymes that install E-double bonds during each addition of a five-carbon isoprene unit . Analysis of the Q99622 and C15 products from incubations with avian P14324 reveals that small amounts of neryl diphosphate ( Z- Q99622 ) and ( Z,E ) -FPP are formed along with the E-isomers during the P01031 --> Q99622 and Q99622 --> C15 reactions . Similar results were obtained for P14324 from Escherichia coli , Artemisia tridentata ( sage brush ) , Pyrococcus furiosus , and Methanobacter thermautotrophicus and for GPP and FPP synthesized in vivo by E. coli P14324 . When ( R ) -[2-2H]IPP was a substrate for chain elongation , no deuterium was found in the chain elongation products . In contrast , the deuterium in ( S ) -[2-2H]IPP was incorporated into all of the products . Thus , the pro-R hydrogen at P06681 of IPP is lost when the E- and Z-double bond isomers are formed . The synthesis of Z-double bond isomers by P14324 during chain elongation is unexpected for a highly evolved enzyme and probably reflects a compromise between optimizing double bond stereoselectivity and the need to exclude DMAPP from the IPP binding site . DB00254 plus tauroursodeoxycholic acid for transthyretin amyloidosis : a phase II study . We designed a phase II , open-label study to evaluate the efficacy , tolerability , safety , and pharmacokinetics of orally doxycycline ( 100 mg P55957 ) and tauroursodeoxycholic acid ( DB08834 ) ( 250 mg three times/day ) administered continuously for 12 months . Primary endpoint is response rate defined as nonprogression of the neuropathy and of the cardiomyopathy . Since July 2010 , we enrolled 20 patients . Seventeen patients have hereditary P02766 , two patients have senile systemic amyloidosis , and one is a domino recipient . Seven patients completed 12-month treatment , 10 completed 6-month treatment , two discontinued because of poor tolerability , and one is lost at follow-up . No serious adverse events were registered . No clinical progression of cardiac involvement was observed . The neuropathy ( Neuropathy Impairment Score in the Lower Limbs [ Q92911 -LL ] and Kumamoto score ) remained substantially stable over 1 year . These preliminary data indicate that the combination of Doxy- DB08834 stabilizes the disease for at least 1 year in the majority of patients with an acceptable toxicity profile . Limited sampling strategies for sirolimus after pediatric renal transplantation . Q86TD4 has been increasingly used in renal transplantation , but limited sampling approaches for estimation of AUC remain elusive . A post-hoc analysis of 94 PK profiles in 75 patients from four previous studies was performed to generate limited sampling approaches for approximation of AUC based on two to four time points for both P55957 and OD Q86TD4 dosing . AUC was calculated using the trapezoid rule . Stepwise linear regression was performed to generate an abbreviated AUC from the limited sampling approaches . For P55957 dosing , complete AUC had a strong correlation with the trough levels ( r(2) = 0.882 , p < 0.0001 ) and with P06681 level ( r(2) = 0.9025 , p < 0.0001 ) . A three-point and a four-point limited sampling approach showed improved agreement with complete AUC compared with single-point sampling . A convenient and accurate ( r(2) = 0.992 ) four-point limited sampling approach reads : AUC = 10; ( 1.085 + 0.117 x log C0 + 0.164 x log C1-0.131 x log P06681 + 0.823 x log C4 ) . Similarly , complete AUC had a statistically significant correlation with the trough levels ( r(2) = 0.549 , p < 0.0001 ) and with P06681 level ( r(2) = 0.716 , p < 0.0001 ) for OD dosing . The estimation of AUC for OD dosing was improved over single-point sampling ( r(2) = 0.951 ) using the formula : AUC = 10; ( 1.100 + 0.115 x log C0 + 0.803 x log C4 ) . This study represented the first limited sampling approach for Q86TD4 . Further studies are required to determine the optimal Q86TD4 target AUC . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC .

MH_train_1040

MH_train_1040

MH_train_1040

interacts_with DB00641?

multiple_choice

Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing P16671 -dependent cholesteryl ester accumulation in macrophages . Protease inhibitors decrease the viral load in HIV patients , however the patients develop hypertriglyceridemia , hypercholesterolemia , and atherosclerosis . It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia . Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of P16671 and the accumulation of cholesteryl esters . The use of P16671 -blocking antibodies , a P16671 morpholino , and monocytes isolated from P16671 null mice demonstrated that protease inhibitor-induced increases in cholesteryl esters were dependent on P16671 upregulation . These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating P16671 and cholesteryl ester accumulation independent of dyslipidemia . Studies with P01130 null mice demonstrated that low doses of protease inhibitors induce an increase in the level of P16671 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids . Furthermore , the lack of P16671 protected the animals from protease inhibitor-induced atherosclerosis . Finally , ritonavir increased P37231 and P16671 mRNA levels in a PKC- and P37231 -dependent manner . We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of P16671 and the subsequent accumulation of sterol in macrophages . Molecular genetic studies in monogenic and polygenic human diseases . The main goal of this study was to determine and characterise the types of mutations in two monogenic human disorders : cystic fibrosis ( CF ) and Duchenne/Becker muscular dystrophy ( P11532 , BMD ) and the susceptibility allele frequency in a polygenic disease : type I insulin-dependent diabetes mellitus ( IDDM ) . After analysing 220 chromosomes for mutations in the CF ( Cystic Fibrosis Transmembrane Conductance Regulator = P13569 ) gene , delta F508 mutation was most abundant ( 41 % ) and out of the non-delta F508 CF mutations 5 % was identified as G542X , G551D , R553X , N1303K and W1282X . The CF haplotype analysis by using linked markers to the P13569 gene revealed that the CF " B " haplotype occurred in 66.7 % of patients , and this haplotype was 57.2 % in patients carrying the delta F508 mutation . Prenatal genetic diagnosis for CF was performed in 10 fetuses : 3 were affected , 6 were carriers , and 1 without any CF mutation . Fifty % of 66 patients with P28068 /BMD muscular dystrophy had one or more exon deletions in the dystrophin gene . Eighty-five % of the deletions occurred at the 3' and 15 % at the 5' end of the gene . Out of the three prenatal diagnosis in one case P11532 was substantiated . Thirty-six % of 50 patients with IDDM possessed four , 44 % three and 20 % two susceptibility markers in the HLA-DQA1 , -DQB1 region . The onset of the disease correlated with the number of susceptibility alleles . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Determination of free N-acetylneuraminic acid in human body fluids by high-performance liquid chromatography with fluorimetric detection . Determinations of both the free and bound form of N-acetyl-neuraminic acid ( NANA ) in several human body fluids , such as serum , cerebrospinal fluid ( P04141 ) , saliva , urine , amniotic fluid , and milk were carried out by HPLC with fluorimetric detection . The method utilized 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride ( P28068 ) as a fluorimetric derivatizing reagent . Free-form NANA was obtained from the body fluids after ultrafiltration with Microcon 10 ( YM-10 cellulose membrane , filtration limit M(r) = 10,000 , Amicon ) . The P28068 derivative of NANA was separated isocratically by a Nucleosil 5C18 column with a mixture of 0.1 M sodium phosphate buffer ( pH 2.0 ) -methanol ( 75:25 , v/v ) . A gradient elution system was used for urine analysis . Analysis times were 10-30 min . Recoveries of free NANA by ultrafiltration were satisfactory : 95.66 +/- 1.80 % for serum and 97.27 +/- 1.55 % for P04141 , respectively . The high sensitivity and specificity render this method applicable to all the body fluids tested . Although a physiological role for free NANA has not yet been elucidated , the method presented promises to contribute to the basic understanding of the NANA metabolism . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Impaired postprandial blood flow in adipose tissue may be an early marker of insulin resistance in type 2 diabetes . OBJECTIVE : We investigated the changes in subcutaneous adipose tissue blood flow ( ATBF ) after a meal in the various stages of type 2 diabetes . RESEARCH DESIGN AND METHODS : Five groups were examined : healthy control subjects , first-degree relatives of subjects with type 2 diabetes , subjects with impaired glucose tolerance ( IGT ) , subjects with type 2 diabetes and postprandial hyperglycemia but normal fasting plasma glucose levels ( diabetes group A [ P28067 ] ) , and subjects with type 2 diabetes with both postprandial and fasting hyperglycemia ( diabetes group B [ P28068 ] ) . ATBF was measured with (133)Xe . RESULTS : ATBF was higher in control subjects ( 1,507 +/- 103 ml/100 cm(3) tissue x min ) versus relatives and IGT , P28067 , and P28068 subjects ( 845 +/- 123 , 679 +/- 69 , 765 +/- 60 , and 757 +/- 69 ml/100 cm(3) tissue x min , respectively ; P < 0.001 ) . P01308 sensitivity index ( ISI ) in control subjects ( 82 +/- 3 mg x l(2)/mmol x mU x min ) was higher versus that for relatives and IGT , P28067 , and P28068 subjects ( 60 +/- 3 , 45 +/- 1 , 40 +/- 6 , and 29 +/- 4 mg x l(2)/mmol x mU x min , respectively ; P < 0.0001 ) . ISI was positively associated with peak-baseline ATBF ( beta coefficient 0.029 +/- 0.013 , P = 0.03 ) . CONCLUSIONS : After meal ingestion , insulin-stimulated ATBF was decreased in relatives and and IGT , P28067 , and P28068 subjects . This defect could be an early marker of insulin resistance that precedes the development of type 2 diabetes . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Metabolic fate of 2,2-dimethylbutyryl moiety of simvastatin in rats : identification of metabolites by gas chromatography/mass spectrometry . Metabolic pathways of simvastatin ( DB00641 ) , a lactone prodrug of an inhibitor of P04035 , were elucidated in male rats , using the [ 14C ] -labelled compound . Evidence has been obtained for hydrolysis of simvastatin and its metabolites at their 2,2-dimethylbutyryl moieties . Metabolites identified in plasma were 2,2-dimethylbutyric acid ( P28068 ) , 2,2-dimethyl-3-hydroxybutyric acid ( DMHB ) and an open chain hydroxy acid of simvastatin : metabolites identified in urine were DMHB , a glucuronide and the glycine conjugate of P28068 . They were characterized by gas chromatography/electron impact and chemical ionization mass spectrometry as phenacyl or pertrimethylsilylated derivatives . The structures of the metabolites and the aglycone of the glucuronide were confirmed as phenacyl esters by comparison of their chromatographic data and mass spectra with those of the phenacyl derivatives of authentic compounds . Activity , pharmacological inhibition and biological regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Trypanosoma brucei . Activity of hydroxymethylglutaryl-coenzyme A ( HMG- DB01992 ) reductase , the key enzyme in the biosynthesis of steroids and polyisoprenoids in mammalian cells , has been detected in both the bloodstream form and the culture-adapted procyclic form of Trypanosoma brucei ( 3.7 +/- 0.6 and 12.7 +/- 1.8 pmol mevalonate produced min-1 ( mg cell protein ) -1 , respectively ) . The enzyme activity is enriched 6-fold in microsomal fractions . Several competitive inhibitors of mammalian P04035 , including synvinolin ( simvastatin ) , inhibit the multiplication of both forms of trypanosome in vitro ( IC50 , approx. 25-50 microM after 2-3 days ) . This growth inhibition is potentiated by agents interfering with the exogenous supply of cholesterol , such as antibodies blocking the low-density lipoprotein ( LDL ) receptor , or 5 microM chloroquine . Conversely , growth inhibition by synvinolin can be largely reverted either by 300 nM LDL or by products of the mevalonate pathway , such as 20 mM mevalonate and in procyclics by 100 microM squalene or cholesterol . In procyclics , low concentrations of synvinolin selectively inhibit the incorporation of [14C]acetate into sterols , but not into fatty acids . These results argue for a critical role in trypanosomes of a mevalonate pathway , that is involved in the biosynthesis of sterol and probably of other metabolites . The P04035 activity is decreased 2-fold in procyclics incubated with 4 mM mevalonate and increased 2-fold in the presence of 2.5 microM synvinolin . DB00641 also upregulates LDL binding up to 4-fold . These data suggest that P04035 and P01130 expression are regulated in T. brucei as in mammalian cells , to ensure sterol homeostasis . UMD ( Universal mutation database ) : a generic software to build and analyze locus-specific databases . The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases . In the near future , the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers , which is growing in frequency in industrial states . The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations . Other fields such as molecular epidemiology will also be developed using these new data . Consequently , the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line . To meet this goal , we devised a generic software called UMD ( Universal Mutation Database ) . It was developed as a generic software to create locus-specific databases ( LSDBs ) with the 4(th) Dimension(R) package from ACI . This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data . Thanks to the flexible structure of the UMD software , it has been successfully adapted to nine genes either involved in cancer ( P25054 , P04637 , P06400 , O00255 , Q09428 , P40337 , and P19544 ) or in genetic diseases ( P35555 and P01130 ) . Four new LSDBs are under construction ( P49748 , P11310 , KIR6 , and P29400 ) . Finally , the data can be transferred to core databases . Molecular determinants of trastuzumab efficacy : What is their clinical relevance ? DB00072 -containing therapy is a standard of care for human epidermal growth factor receptor-2 ( P04626 ) -positive breast cancer . In pre-clinical models , a wide range of molecular mechanisms have been associated with reduced sensitivity to trastuzumab in vitro . These include expression of the truncated P04626 receptor fragment p95HER2 , activating mutation of the gene encoding the class 1A catalytic subunit of phosphatidylinositol 3-kinase ( P42336 ) , loss of phosphatase and tensin homolog ( P60484 ) , activation of other downstream signal transducers , prevention of cell cycle arrest , increased signaling through alternative ( HER or non-HER ) tyrosine kinase receptors , and resistance to antibody-dependent cellular cytotoxicity . However , the clinical significance of these mechanisms as determinants of trastuzumab efficacy in vivo has been unclear . Here , we review clinical studies of potential predictive biomarkers of trastuzumab efficacy in P04626 -positive breast cancer and consider whether evaluation of such markers might inform patient selection for therapy . We find that clinical evidence relating to potential predictive biomarkers is mostly limited to small , retrospective studies , many of which have yielded conflicting findings . Some trends are evident in the retrospective data and in biomarker analyses from randomized clinical trials , particularly relating to activation of the phosphatidylinositol 3-kinase pathway , but none is sufficiently strong to form a basis for patient selection . This may be explained by the fact that multiple mechanisms of action determine the clinical efficacy of trastuzumab . In the absence of novel , validated biomarkers of efficacy , trastuzumab eligibility should continue to be based on evaluation of P04626 status according to standard methods . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Assignment of eight loci to bovine syntenic groups by use of PCR : extension of a comparative gene map . The polymerase chain reaction ( PCR ) has been combined with hybrid somatic cell technology to extend the bovine physical map . Eight bovine loci -- glycoprotein hormone alpha ( P01215 ) , coagulation factor X ( F10 ) , chromogranin A ( P10645 ) , low-density lipoprotein receptor ( P01130 ) , human prochymosin pseudogene ( P23946 ) , oxytocin ( P01178 ) , arginine-vasopressin ( P01185 ) , and cytochrome oxidase c subunit IV pseudogene ( COXP ) -- were assigned to bovine syntenic groups with this approach . P01215 was assigned to bovine syntenic group U2 , F10 to U27 , P10645 to U4 [ bovine Chromosome ( Chr ) 21 ] , P01130 to U22 , P23946 to U6 , P01178 and P01185 to U11 , and COXP to U3 ( bovine Chr 5 ) . Seven of these genes , P01215 , F10 , P10645 , P01130 , P01178 , P01185 , and P23946 , further delineate regions of chromosomal conservation on human Chrs 6 , 13 , 14 , 19 , 20 , 20 , and 1 , respectively . P10645 , P01178 , and P01185 are unmapped in the mouse . Comparative mapping predicts the mouse P10645 will map to Chr 12 , and mouse P01178 and P01185 will map to mouse Chr 2 . Furthermore , human P23946 is predicted to be sublocalized to 1p32-q21 . The primers developed for these eight loci will be useful for the development of hybrid somatic cell panels in the future as well as establishing a collection of bovine expressed sequence tags . Increased plasma thyroid hormone concentrations in P01130 deficient mice may be explained by inhibition of aryl hydrocarbon receptor-dependent expression of hepatic UDP-glucuronosyltransferases . BACKGROUND : Overexpression of P36956 causes a repression of hepatic genes involved in phase II metabolism . In P01130 deficient ( P01130 (-/-) ) mice , active levels of P36956 in the liver are increased . We investigated the hypothesis that P01130 (-/-) mice have increased concentrations of thyroid hormones in plasma due to a reduced hepatic glucuronidation . METHODS : Female P01130 (-/-) and wild-type mice were used to study the effect of the P01130 (-/-) genotype on thyroid hormone metabolism . RESULTS : P01130 (-/-) mice had a higher concentration of nuclear P36956 , higher concentrations of thyroxine and triiodothyronine in plasma , a lower expression of relevant P22309 isoforms , reduced activities of pNP- P78381 , T(3)- P78381 and T(4)- P78381 and a lower mRNA and protein concentration of P35869 in the liver than wild-type mice ( P < 0.05 ) . Plasma concentration of DB00024 , mRNA concentrations of various genes involved in thyroid hormone synthesis in the thyroid , activity of deiodinase and mRNA concentrations of two thyroid hormone responsive genes , P22680 and Na(+)/K(+)-ATPase , in the liver did not differ between both genotypes . CONCLUSIONS : This study shows that P01130 (-/-) mice have increased concentrations of thyroid hormones in plasma . This effect is probably due to an inhibition of thyroid hormone glucuronidation , which might be caused by down-regulation of P78381 genes due to a reduced expression of P35869 . However , with respect to plasma DB00024 concentration and expression of thyroid hormone responsive genes no overt hyperthyroidism was detected . GENERAL SIGNIFICANCE : P01130 deficiency leads to a reduced glucuronidation of thyroid hormones in the liver which causes a moderate increase of plasma thyroid hormone concentrations . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . Gorlin syndrome and desmoplastic medulloblastoma : Report of 3 cases with unfavorable clinical course and novel mutations . We present three cases of genetically confirmed Gorlin syndrome with desmoplastic medulloblastoma ( P28068 ) in whom tumor recurred despite standard therapy . One patient was found to have a novel germline missense Q13635 mutation . Molecular analysis of recurrent tumor using fluorescent in situ hybridization ( Q5TCZ1 ) revealed P60484 and/ or Q13635 loss in 2 patients . Whole exome sequencing ( WES ) of tumor in one patient revealed loss of heterozygosity of Q13635 and a mutation of GNAS gene in its non-coding 3' -untranslated region ( UTR ) with corresponding decreased protein expression . While one patient died despite high-dose chemotherapy ( HDC ) plus stem cell rescue ( P04156 ) and palliative radiotherapy , two patients are currently alive for 18+ and 120+ months respectively following retrieval therapy that did not include irradiation . Infants with P28068 and GS should be treated aggressively with chemotherapy at diagnosis to prevent relapse but radiotherapy should be avoided . The use of molecular prognostic markers for P28068 should be routinely used to identify the subset of tumors that might have an aggressive course . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Genetic influences on sarcoidosis . To investigate the genetic influences underlying the development of sarcoidosis , HLA class II genotyping was performed in Japanese patients with sarcoidosis and healthy controls using the PCR-RFLP method . The frequencies of both DR52 group antigen-associated alleles ( HLA- Q8IUH3 *11 , - Q8IUH3 *12 and - Q8IUH3 *14 ) and Q8IUH3 *08 alleles were higher in the patient group , suggesting that the common , specific amino acid residue on the Q8IUH3 molecule of these alleles may determine susceptibility to sarcoidosis . Alternatively , it is possible that another susceptibility gene , linked to these Q8IUH3 alleles , exists within the MHC region . We screened the P01375 , P01374 , P0DMV8 and Hum70t genes around the class III region , as well as the P28067 and - P28068 genes in the class II region , for genetic polymorphism in sarcoidosis . None of these genes suggested a susceptibility to sarcoidosis . These studies support the thesis that one of the major genetic factors controlling the development of sarcoidosis is located within the Q8IUH3 locus in the HLA class II region . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors .

MH_train_1041

MH_train_1041

MH_train_1041

interacts_with DB00945?

multiple_choice

Targeting cyclooxygenase-2 with sodium butyrate and NSAIDs on colorectal adenoma/carcinoma cells . AIM : The protective effects of sodium butyrate and NSAIDs ( especially the highly selective P35354 inhibitors ) have attracted considerable interest recently . In this study , primary adenoma cells and HT-29 were used to investigate whether the above drugs would be effective for reducing proliferation and inducing apoptosis . Additionally , it was investigated whether NSAIDs would strengthen the effects of sodium butyrate and its possible mechanisms . METHODS : In vitro primary cell culture of colorectal adenomas and HT-29 were used for this investigation . DB00917 isolated from HT-29 cell culture supernatants was investigated by ELISA . MTT was employed to detect the anti-proliferative effects on both adenoma and HT-29 culture cells . DB00828 was used for apoptosis rate and cell cycle analysis . The morphology of apoptotic cells was investigated by means of electromicroscopy . RESULTS : Sodium butyrate could stimulate the secretion of DB00917 , while NSAIDs inhibited it to below 30 pg/10(6) cells . Both butyrate and NSAIDs could inhibit cell proliferation and induce apoptosis . The effects were time- and dose-dependent ( P < 0.05 ) . DB00945 and NS-398 could enhance the effects of sodium butyrate . The effects were stronger while sodium butyrate was used in combination with NS-398 than it was used in combination with aspirin . CONCLUSION : Butyrate and NSAIDs could inhibit cell proliferation and induce apoptosis respectively . NSAIDs could enhance the effects of sodium butyrate by down-regulating P35354 expression . Selective P35354 inhibitor is better than traditional NSAIDs . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . DB00945 and non-steroidal anti-inflammatory drugs for cancer prevention : an international consensus statement . Evidence clearly shows a chemopreventive effect for aspirin and other non-steroidal anti-inflammatory drugs ( NSAIDs ) on colorectal cancer and probably other cancer types ; however , data on the risk-benefit profile for cancer prevention are insufficient and no definitive recommendations can be made . DB00945 has emerged as the most likely NSAID for use in chemoprevention because of its known cardiovascular benefit and available safety and efficacy data . Other traditional NSAIDs , particularly sulindac , and selective P35354 inhibitors are now given to patients at high risk of colorectal cancer , although these drugs do not provide cardioprotection . More studies of aspirin and cancer prevention are needed to define the lowest effective dose , the age at which to initiate therapy , the optimum treatment duration , and the subpopulations for which the benefits of chemoprevention outweigh the risks of adverse side-effects . Although it might be possible to answer some of these questions with longer follow-up of existing clinical trials , randomised controlled trials with new study designs will be needed . Future projects should investigate the effects of aspirin treatment on multiple organ systems . Cancers of interest are colorectal , breast , prostate , lung , stomach , and oesophageal . The main side-effect of aspirin is peptic ulcers ; therefore coadministration of aspirin with a proton-pump inhibitor is an attractive option and is under investigation in the AspECT trial . DB00945 , ibuprofen , and other non-steroidal anti-inflammatory drugs in cancer prevention : a critical review of non-selective P35354 blockade ( review ) . DB00945 in the 21st century-common mechanisms of disease and their modulation by aspirin : a report from the 2015 scientific conference of the international aspirin foundation , 28 August , London , UK . Professor Peter Rothwell of Oxford University chaired the annual Scientific Conference of the International DB00945 Foundation in London on 28 August 2015 . It took the form of four sessions . DB00945 has more than one action in its effects on disease . Its acetylation of cyclooxygenase 2 ( P35354 ) in platelets leads to the blockade of pro-inflammatory chemicals and generation of anti-inflammatory mediators and increase in nitrous oxide ( NO ) production , which helps to preserve arterial endothelium . But platelets are not its only target . There is now evidence that aspirin has a direct antitumour effect on intestinal mucosal cells that block their potential transformation into cancer cells . Randomised placebo-controlled trials ( RCTs ) in people with histories of colorectal neoplasia have shown that aspirin reduces the risk of recurrent adenomas and reduces long-term cancer incidence in patients with Lynch syndrome . Among women given aspirin for cardiovascular disease , there were fewer cancers than in those given placebo . Epidemiological evidence has suggested that aspirin treatment after cancer is diagnosed reduces the incidence of metastases and prolongs survival , and long-term studies of anticancer treatment with aspirin are under way to confirm this . Apart from cancer studies , aspirin use is now firmly established as treatment for antiphospholipid syndrome ( Hughes syndrome ) and is being used to prevent and treat the heightened risk of cardiovascular disease in diabetes mellitus and in patients with HIV . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . P02751 -induced P35354 mediates P08253 expression and invasiveness of rhabdomyosarcoma . Although accumulating evidence suggests the importance of cyclooxygenase-2 ( P35354 ) and prostaglandin E(2) ( PGE(2) ) in the pathogenesis of many cancers , the mechanism by which this enzyme and its metabolite promote cancer progression is unknown . In this study , we investigated the role of P35354 in fibronectin-induced up-regulation of rhabdomyosarcoma matrix metalloproteinase ( MMP ) -2 activity and cellular invasiveness . We tested three human rhabdomyosarcoma cell lines : RMS559 , RD , and SJRH30 . Cell attachment to fibronectin up-regulated both P35354 expression and PGE(2) production and concomitantly enhanced P08253 activity . Exogenous PGE(2) stimulated P08253 promoter activity , increased P08253 expression , and increased cellular invasiveness . DB00945 and rofecoxib ( non-selective and selective P35354 inhibitor , respectively ) each abolished fibronectin-associated induction of P08253 and induced dose-dependent reductions in cellular invasiveness . These data implicated a role for inducible P35354 and PGE(2) in the regulation of rhabdomyosarcoma cellular invasiveness and P08253 activity . The effect of cilostazol and aspirin pre-treatment against subsequent transient focal cerebral ischemia in rat . OBJECTIVES : Among several anti-platelet drugs to prevent recurrent stroke , cilostazol has shown various effects besides its anti-platelet activity . We examined whether 7 days of oral administration of cilostazol protects against subsequent cerebral ischemia , and whether or not the effect of combination therapy with aspirin is more protective . METHODS : We used Sprague-Dawley ( SD ) rats and assigned them to four groups : vehicle , aspirin , cilostazol , and aspirin plus cilostazol combination therapy . After oral administration of anti-platelets for 7 days , we performed transient middle cerebral artery occlusion ( MCAO ) for 90 minutes , and examined infarct volume , neurological symptoms , and regional cerebral blood flow ( rCBF ) . Immunostaining of Bax , Bcl-2 , TUNEL , 4-HNE , 8-OHdG , and P35354 was performed 24 hours after ischemia . RESULTS : The cilostazol group and the combination therapy group showed significant decreases of infarct volume and significant improvements of rCBF during ischemia , compared with the vehicle or aspirin group . Significant decreases of Bax , TUNEL , 8-OHdG , and 4-HNE expression in the combination therapy group , compared with those in the vehicle or aspirin group , were shown in the boundary zone . P35354 expression was unexpectedly increased in the combination therapy group . DISCUSSION : DB00945 co-administration did not inhibit this effect . The addition of the oral administration of cilostazol either alone or with aspirin administration may be beneficial for subsequent cerebral ischemic damage in terms of reducing infarct volume , improving rCBF during ischemia , inhibiting the apoptotic pathway , and reducing oxidative stress . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Genetic mechanism of aspirin-induced urticaria/angioedema . PURPOSE OF REVIEW : DB00945 -induced urticaria/angioedema is a major aspirin-related hypersensitivity often associated with aspirin-intolerant asthma . Genetic studies on aspirin-intolerant asthma have shown chronic overproduction of cysteinyl leukotrienes . The genetic analysis of aspirin-induced urticaria/angioedema is limited , however . RECENT FINDINGS : A recent study on HLA genotypes has suggested that the HLA alleles DRB11302 and DQB10609 may be genetic markers for aspirin-induced urticaria/angioedema . A polymorphism study that examined nine single-nucleotide polymorphisms of five leukotriene-related genes [ P09917 ( encoding P09917 ) , P20292 ( P09917 -activating protein ) , P35354 ( cyclooxygenase 2 ) , Q16873 ( leukotriene C4 synthase ) , and Q9Y271 ( cysteinyl leukotriene receptor 1 ) ] found that promoter polymorphisms of P09917 ( -1708A > G ) and Q9Y271 ( -634C > T ) were significantly different between aspirin-intolerant asthma and aspirin-induced urticaria/angioedema , suggesting different contributions to the lipoxygenase pathway . A second polymorphism study , conducted on histamine-related genes , did not find any significant associations with aspirin-induced urticaria/angioedema for the genes P50135 ( encoding histamine N-methyltransferase ) , P35367 or P25021 ( encoding histamine receptor types 1 and 2 respectively ) , or the gene encoding high-affinity IgE receptor Ibeta ( FcepsilonRIbeta ) ; however , the FcepsilonRIalpha gene promoter polymorphism was significantly associated with aspirin-induced urticaria/angioedema . This finding has been supported by in vitro functional studies . SUMMARY : The HLA alleles DRB11302 and DQB10609 , and the P09917 and FcepsilonRIalpha promoter polymorphisms , may contribute to the pathogenesis of aspirin-induced urticaria/angioedema . Further investigation to identify candidate genetic markers would help to elucidate the pathogenic mechanism of this condition . P35354 inhibitor ( SC-236 ) suppresses activator protein-1 through c-Jun NH2-terminal kinase . BACKGROUND AND AIMS : DB00945 exerts antitumor effect partly through blocking tumor promoter-induced activator protein-1 ( AP-1 ) activation . The aim of this study is to determine how specific P35354 inhibitor SC-236 mediates antitumor effect by modulation of AP-1-signaling pathway . METHODS : AP-1 transcriptional activity and DNA-binding activity were detected by luciferase reporter assay and gel shift assay , separately . Mitogen-activated protein kinase ( MAPK ) activation was determined by Western blot and in vitro kinase assay . Antisense oligonucleotide against c-Jun-N-terminal kinase ( JNK ) was used to suppress JNK expression . RESULTS : We showed that SC-236 inhibited 12-O-tetradecanoylphorbol-13-acetate ( PMA ) -induced cell transformation in a dose-dependent manner in JB6 cells . At a dose range ( 12.5-50 micromol/L ) that inhibited cell transformation , SC-236 also inhibited anchorage-independent cell growth and AP-1-activation in 3 gastric cancer cells , independent of P36551 -prostaglandin synthesis . SC-236 down-regulated c-Jun-NH2-terminal kinase phosphorylation and activity . Suppression of JNK activity reversed the inhibitory effect on AP-1 activity by SC-236 and suppressed gastric cancer cell growth , indicating that the inhibitory effect of SC-236 on AP-1 activation and cell growth was through interaction with JNK . CONCLUSIONS : The inhibitory effect on JNK-c-Jun/AP-1 activation contributes to the antitumor effect of P35354 -specific inhibitor , and inhibition of JNK activation may have a therapeutic benefit against gastric cancer . Dependence on phosphoinositide 3-kinase and DB01367 -RAF pathways drive the activity of RAF265 , a novel RAF/ P35968 inhibitor , and RAD001 ( DB01590 ) in combination . Activation of phosphatidylinositol-3-kinase ( PI3K ) -AKT and Kirsten rat sarcoma viral oncogene homologue ( P01116 ) can induce cellular immortalization , proliferation , and resistance to anticancer therapeutics such as epidermal growth factor receptor inhibitors or chemotherapy . This study assessed the consequences of inhibiting these two pathways in tumor cells with activation of P01116 , PI3K-AKT , or both . We investigated whether the combination of a novel RAF/vascular endothelial growth factor receptor inhibitor , RAF265 , with a mammalian target of rapamycin ( P42345 ) inhibitor , RAD001 ( everolimus ) , could lead to enhanced antitumoral effects in vitro and in vivo . To address this question , we used cell lines with different status regarding P01116 , P42336 , and P15056 mutations , using immunoblotting to evaluate the inhibitors , and MTT and clonogenic assays for effects on cell viability and proliferation . Subcutaneous xenografts were used to assess the activity of the combination in vivo . RAD001 inhibited P42345 downstream signaling in all cell lines , whereas RAF265 inhibited RAF downstream signaling only in P15056 mutant cells . In vitro , addition of RAF265 to RAD001 led to decreased AKT , S6 , and P06730 binding protein 1 phosphorylation in HCT116 cells . In vitro and in vivo , RAD001 addition enhanced the antitumoral effect of RAF265 in HCT116 and H460 cells ( both P01116 mut , P42336 mut ) ; in contrast , the combination of RAF265 and RAD001 yielded no additional activity in A549 and MDAMB231 cells . The combination of RAF and P42345 inhibitors is effective for enhancing antitumoral effects in cells with deregulation of both DB01367 -RAF and PI3K , possibly through the cross-inhibition of 4E binding protein 1 and S6 protein . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Nonsteroidal anti-inflammatory drugs : adverse effects and their prevention . OBJECTIVES : To discuss nonsteroidal anti-inflammatory drugs ( NSAIDs ) , their history , development , mode of action , toxicities , strategies for the prevention of toxicity , and future developments . METHODS : Medline search for articles published up to 2007 , using the keywords acetylsalicylic acid , aspirin , NSAIDs , cyclooxygenase 2 , adverse effects , ulcer , and cardiovascular . RESULTS : NSAIDs are 1 of the oldest , most successful drugs known to modern medicine . They are effective for alleviating pain , fever , and inflammation by inhibiting prostaglandin synthesis . DB00945 , by its irreversible inhibition of blood platelet function , is also effective in the prevention of cardiovascular disease . NSAIDs may cause gastrointestinal ulcers , serious cardiovascular events , hypertension , acute renal failure , and worsening of preexisting heart failure . These adverse effects may be prevented by limiting NSAID dosage and duration and by performing individual risk assessments and treating patients accordingly . Those at risk for gastroduodenal ulcers may be treated with concomitant proton-pump inhibitors , misoprostol and/or P35354 selective NSAIDs . Those at risk for cardiovascular events may be treated with naproxen and a proton-pump inhibitor or misoprostol , but should best avoid NSAID use altogether . CONCLUSIONS : Physicians should always prescribe the lowest effective dose for the shortest possible time and must take into account both the gastrointestinal and the cardiovascular risks of individual patients when prescribing NSAIDs . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . DB00945 inhibits P14780 mRNA expression and release via the PPARalpha/gamma and P35354 /mPGES-1-mediated pathways in macrophages derived from THP-1 cells . In present study , we investigated the effects of aspirin on matrix metalloproteinase ( MMP ) -9 mRNA expression and release and its possible mechanisms in macrophages derived from THP-1 cells . The macrophages were divided into different groups and treated with different drugs , the mRNA expression of P14780 , peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma , cyclooxygenase ( P36551 ) -2 , membranebound prostaglandin E synthase ( mPGES ) -1 in macrophages were examined with reverse-transcription polymerase chain reaction , and the protein expressions of Q07869 alpha and gamma , mPGES-1 were detected by Western-blot , the levels of P14780 and PGE(2) in cultured supernatants were determined with enzyme-linked immunosorbent assay . The results indicated that after the macrophages were incubated with aspirin for 24h , the P14780 mRNA expression and release were decreased , while the Q07869 alpha/gamma mRNA and protein expression was increased , respectively , and Q07869 alpha/gamma agonists could also decrease P14780 mRNA expression and release . Additionally , the P35354 mRNA expression , mPGES-1 mRNA and protein expression in macrophages were all decreased after incubation with aspirin for 24h and the PGE(2) release was also decreased . The macrophages stimulated with PGE(2) for 24h might increase the P14780 mRNA expression and release . When PGE(2) plus Q07869 alpha agonist or Q07869 gamma agonist were simultaneously used , the stimulation of P14780 mRNA expression and release by PGE(2) was significantly decreased . It might be concluded that aspirin could inhibit the P14780 gene expression and release through the PPARalpha/gamma and P35354 /mPGES-1-mediated pathways and the two pathways might be partly overlapped and even be interrelated . Exploring the potential chemopreventative effect of aspirin and rofecoxib on hereditary nonpolyposis colorectal cancer-like endometrial cancer cells in vitro through mechanisms involving apoptosis , the cell cycle , and mismatch repair gene expression . Women in hereditary nonpolyposis colorectal cancer ( HNPCC ) families have up to a 71 % lifetime risk for developing endometrial cancer ( EC ) . This compares to the female lifetime risk for colorectal cancer ( CRC ) in HNPCC of 60 % . The basis of HNPCC is an inherited mutation in a mismatch repair gene ( P22897 ) . DB00945 and P35354 inhibitors seem to have a chemoprotective effect on CRC in the general population and are the subject of prospective clinical studies in patients at high risk for CRC including HNPCC . There is no evidence that these agents have any protective effect against EC in the general population . This study investigated the effect of aspirin and a P35354 inhibitor ( rofecoxib ) on an HNPCC EC cell line model ( Ishikawa ) by assessing the effect on proliferation , apoptosis , the cell cycle , and P22897 gene expression . DB00945 inhibits EC cell proliferation by inducing apoptosis and changes in the cell cycle . This effect is not mediated by changes in P22897 gene ( P43246 ) expression as assessed by quantitative reverse transcription-polymerase chain reaction . DB00533 inhibits EC cell proliferation ; this did not appear to be mediated by induction of apoptosis , by alterations of the cell cycle , or by changes in P22897 gene expression . Importance of surface properties of affinity resin for capturing a target protein , cyclooxygenase-1 . We have prepared affinity resins based on two kinds of solid phases , including a commercially available solid phase , to re-realize the importance of surface properties of affinity resins such as controlled ligand density as well as existential surroundings of the ligand . Affinity resins were prepared using non-steroidal anti-inflammatory drugs , such as Ketoprofen , Ibuprofen , and DB00945 , having different activities as ligands . The ligand density was controlled through two different strategies : one strategy was that the solid phases having different amino group densities ( 20 , 60 , 100 , 125 micromol/ml ) were utilized then , Ketoprofen was fully immobilized through condensation reaction to amino groups ; another strategy was that a solid phase having amino group density ( 125 micromol/ml ) was utilized then , each ligand was immobilized with controlled immobilization rate . In addition , a typical hydrophobic group , stearoyl group ( C(18) group ) , was immobilized on the affinity resin with controlled ligand immobilization rate to change the existential surroundings of the ligand . Affinity tests were performed for Cyclooxgenase-1 ( P23219 ) as it was the target protein in this work . The amount of captured P23219 was evaluated utilizing each affinity resin . It was suggested that the density of surface ligand tends to relate to the amount of captured P23219 on our solid phase-based affinity resins ; however , several exceptions occurred according to the surface properties of affinity resins in the case of commercial one . P35354 promotes early atherosclerotic lesion formation in ApoE-deficient and C57BL/6 mice . Cyclooxygenase ( P36551 ) 2 is expressed in atherosclerotic lesions . We have previously reported that selective inhibition of P35354 reduces early atherosclerosis in P01130 deficient mice . To examine the role of P35354 in atherosclerosis in other mouse models , we studied the effects of selective P35354 inhibition ( by rofecoxib and NS-398 ) and nonselective P36551 inhibition ( by indomethacin ) on early atherosclerotic lesion formation in apolipoprotein E-deficient ( apoE(-/-) ) mice . Selective P35354 and nonselective P36551 inhibition reduced atherosclerosis in female apoE(-/-) mice by 35-38 % and 38-51 % in the proximal and en face aortas , respectively . Next we investigated the role of macrophage P35354 by transplanting P35354 (-/-) fetal liver cells into C57BL/6 mice and challenging the mice with an atherogenic diet . Genetic deletion of P35354 from hematopoietic cells reduced atherosclerosis by 51 % . In addition , LPS activated P35354 (-/-) macrophages had decreased expression of monocyte chemoattractant protein-1 ( P13500 ) and tumor necrosis factor-alpha ( TNFalpha ) . The results demonstrate that selective inhibition of P35354 and elimination of P35354 from macrophages significantly reduces early atherosclerotic lesion formation in apoE-deficient and C57BL/6 mice . These results are compatible with P35354 expression by macrophages having a proatherogenic role , and support the potential of anti-inflammatory therapeutic approaches for atherosclerosis . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . Diagnosis , prevention and treatment of aspirin-induced asthma and rhinitis . Bronchial asthma is not a homogenous disease . Several variants of asthma can be distinguished . One of them is aspirin-induced asthma . In this distinct clinical syndrome , aspirin and most other nonsteroidal anti-inflammatory drugs that inhibit cyclooxygenase-1 precipitate rhinitis and asthma attacks . This type of asthma affects 5-10 % of adult asthmatics , but remains largely underdiagnosed . The natural history of aspirin-induced asthma ( AIA ) has been described , based on an extensive pan-European survey . DB00945 provocation tests with improved diagnostic accuracy have been developed , although no in-vitro tests has been found to be of diagnostic value . Recent interest in AIA has been stirred by the finding of alterations in arachidonate metabolic pathways , leading to cysteinyl-leukotriene overproduction . Q16873 is overexpressed in bronchi and its mRNA is upregulated in peripheral blood eosinophils . The gene coding for Q16873 exists in two common alleles , one of which appears to be associated with a severe , steroid-dependent type of asthma . New highly specific P35354 inhibitors appear to be a safe alternative for patients with aspirin-induced asthma . Mediator subunit Gal11p/ Q96RN5 is required for fatty acid-dependent gene activation by yeast transcription factor Oaf1p . The yeast zinc cluster transcription factor Oaf1p activates transcription of target genes in response to direct binding of fatty acids in a manner analogous to the vertebrate nuclear receptor peroxisome proliferator-activated receptoralpha ( PPARalpha ) . PPARs and other metazoan nuclear receptors productively engage several distinct LXXLL motif-containing co-activators , including P52701 family members and the Q15648 /MED1 subunit of the Mediator co-activator , to promote ligand-dependent gene activation . Yeast , however , does not appear to harbor LXXLL motif co-activators , and the mechanism of fatty acid-dependent gene activation by the yeast PPARalpha analog Oaf1p is unknown . Here we show that the yeast Mediator subunit Gal11p/ Q96RN5 and its activator-targeted KIX domain plays a critical role in fatty acid-dependent transcriptional regulation of fatty acid beta-oxidation and peroxisomal genes by Oaf1p and for the ability of yeast to utilize fatty acids as a sole carbon source . Moreover , structural studies by NMR spectroscopy reveal that the Oaf1p activation domain interacts with the Gal11p/ Q96RN5 KIX domain in a manner similar to the yeast zinc cluster family member and xenobiotic receptor Pdr1p , revealing that the Gal11p/ Q96RN5 KIX domain is a key target of several ligand-dependent transcription factors in yeast . Together with previous work showing that the Caenorhabditis elegans Gal11p/ Q96RN5 homolog MDT-15 plays a critical role in regulation of fatty acid metabolism by the nematode Q07869 -like nuclear receptor NHR-49 , the findings presented here provide evidence for an ancient and essential role of a Mediator co-activator subunit in regulation of fatty acid metabolism by nuclear receptor-like transcription factors in eukaryotes . DB00945 regulates expression and function of scavenger receptor-BI in macrophages : studies in primary human macrophages and in mice . Scavenger receptor class B type I ( Q8WTV0 ) has been shown to be expressed in human atherosclerotic plaque macrophages , where it is believed to reduce atherosclerosis by promoting cholesterol efflux . In this study we investigated the influence of aspirin and other NSAIDs on Q8WTV0 expression and function in cultured human macrophages as well as in different mouse strains . Incubation of human macrophages with 0.5 mmol/l aspirin resulted in increased Q8WTV0 protein expression and increased uptake of HDL-associated [3H]cholesteryl oleate without changes of Q8WTV0 mRNA levels . In contrast , using 5 mmol/l of aspirin , Q8WTV0 expression and function were significantly decreased . Sodium salicylate exerted similar effects on Q8WTV0 expression , whereas no effects were observed using known P23219 /2 inhibitors ibuprofen and naproxen , respectively . In in vivo studies low-dose aspirin treatment ( 6 mg/kg.day ) induced Q8WTV0 expression in wild-type and Q07869 knockout mice , respectively , whereas the opposite effect was observed upon high-dose aspirin treatment ( 60 mg/kg.day ) in these animals . We could show that P36551 -independent effects of aspirin were able to enhance expression of Q8WTV0 in macrophages in a post-transcriptional , Q07869 independent way , suggesting a novel pharmacologic effect of aspirin . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Topical glucocorticoids downregulate P23219 positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 ) are key enzymes in the synthesis of both pro- ( P23219 , P35354 ) and anti-inflammatory prostanoids ( P35354 ) , we investigated the role of topical GC on P23219 , P35354 and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 , CD117 , MBP , elastase , IgE , BB-1 , P05112 , P05113 and P05231 ) , P23219 and P35354 was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 + cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 + NP cells ( P < 0.05 ) , but have no significant effect on P35354 + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 + cells is however increased significantly upon GC treatment ( P < 0.05 ) . Cox-2 inhibitor attenuates NO-induced P29475 in rat caudal trigeminal nucleus . OBJECTIVE : The aim of the present study was to determine which isoform of the cyclooxygenase ( P36551 ) enzyme plays a role in the neuronal nitric oxide synthase ( P29475 ) activation caused by nitroglycerin ( NTG ) , in the most caudal part of the trigeminal caudal nucleus ( P24821 ) of the rat . BACKGROUND : DB00435 donor , NTG , can trigger migraine attack in migraineurs , but not in healthy persons . In rats , subcutaneous administration of NTG ( 10 mg/kg ) increases significantly the number of P29475 -immunoreactive neurons in the P24821 after 4 hours , which could be attenuated by acetyl-salicylate ( DB00945 ) , a nonselective P36551 -inhibitor . METHODS : SPRD rats were divided into 3 groups : ( 1 ) control group ( no drug administration ) , ( 2 ) NS398 ( selective P35354 inhibitor ) administration ( 1 , 3 , or 5 mg/kg ) , and ( 3 ) SC560 ( selective P23219 inhibitor ) administration ( 1 , 5 , or 10 mg/kg ) . Thirty minutes after drug administration , the animals received NTG ( 10 mg/kg ) or placebo injection . Four hours later the animals were transcardially perfused and the cervical part of the P24821 was removed for immunohistochemistry . Results.-The selective P35354 inhibitor NS398 in contrast to the selective P23219 inhibitor SC560 attenuates the NTG-induced P29475 expression dose-dependently . CONCLUSION : These findings suggest that metabolites deriving from P35354 ( but not P23219 ) may be the most important factors in the NTG-induced P29475 expression . These data could help to better understand the pathogenesis of headaches and the action of antimigraine drugs . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . DB00945 induces apoptosis in oesophageal cancer cells by inhibiting the pathway of NF-kappaB downstream regulation of cyclooxygenase-2 . BACKGROUND : DB00945 has potential in the prevention or treatment of oesophageal cancer , the seventh most common cancer in the world , but its mechanism of action is still not certain . METHODS : The oesophageal squamous cell carcinoma cell line TE-13 was cultured with aspirin at different concentrations or for different times . Proliferation and apoptosis were measured by MTT reduction and flow cytometry . Expression of P35354 mRNA was measured by RT-PCR and P35354 protein levels with Western blot analysis . Nuclear NF-kappaB and cytoplasmic IkappaB protein levels were determined by electrophoretic mobility shift assay and Western blot , respectively . RESULTS : DB00945 significantly inhibited cell proliferation and induced apoptosis at concentrations of 1 , 4 , 8 mmol/L . DB00945 dose-dependently decreased the levels of P35354 mRNA , P35354 protein and nuclear NF-kappaB protein and increased the cytoplasmic IkappaB protein . CONCLUSION : We conclude that aspirin inhibits the proliferation of , and induced apoptosis in , the cultured TE-13 SCC cell line . These changes correlate with a reduction in P35354 mRNA and protein expression , prostaglandin synthesis , an inhibition of NF-kappaB nuclear translocation , and an increase in cytoplasmic IkappaB . These results support the further investigation of the cyclooxygenase pathway in investigating the potential of aspirin and similar drugs in cancer prevention and therapy . The database dbEST correctly predicts gene expression in colon cancer patients . This study aims to test the predictive power of gene expression data derived from NIH 's database dbEST , which collects gene expression results from a large number and variety of DNA array experiments . The motivation of this study is to make comparable experimental studies , which are usually performed only for one or a few tissues or organs , with a wide variety of other tissues . Confirmation of a good predictive power of dbEST would put a number of interesting and partially surprising recent findings , solely based on data mining , on a more solid basis than available so far . The expression of nine genes ( P06730 , P26196 , O14929 , Q96RU2 , HSP90 ( beta , P14618 , P53350 , P35354 and P10451 ) plus two calibration genes in paired normal and cancer colon tissues of eight individual patients was investigated by quantitative RT-PCR and compared with the predictions made by the data-base . GUS and beta-actin reveal only little variation among different patients , making them good internal calibration standards . In normal colon tissue , data mining correctly predicts the expression of all nine genes , which covers two orders of magnitude . In cancer , dbEST is somewhat less precise , but still valuable for the comparison with clinical results . Novel agents that potentially inhibit irinotecan-induced diarrhea . DB00762 ( CPT-11 , 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin ) has exhibited clinical activities against a broad spectrum of carcinomas by inhibiting P11387 ( Topo I ) . However , severe and unpredictable dosing-limiting toxicities ( mainly myelosuppression and severe diarrhea ) hinder its clinical use . The latter consists of early and late-onset diarrhea , occurring within 24 hr or > or = 24 hr after CPT-11 administration , respectively . This review highlights novel agents potentially inhibiting CPT-11-induced diarrhea , which are designed and tested under guidance of disposition pathways and potential toxicity mechanisms . Early-onset diarrhea is observed immediately after CPT-11 infusion and probably due to the inhibition of acetylcholinesterase activity , which can be eliminated by administration of atropine . Late-onset diarrhea appears to be associated with intestinal exposure to SN-38 ( 7-ethyl-10-hydroxycamptothecin ) , the major active metabolite of CPT-11 , which may bind to Topo I and induce apoptosis of intestinal epithelia , leading to the disturbance in the absorptive and secretory functions of mucosa . CPT-11 and SN-38 may also stimulate the production of pro-inflammatory cytokines and prostaglandins ( PGs ) , thus inducing the secretion of Na(+) and Cl(-) . Early treatment of severe late-onset diarrhea with oral high-dose loperamide has decreased patient morbidity . Extensive studies have been conducted to identify other potential agents to ameliorate diarrhea in preclinical and clinical models . These include intestinal alkalizing agents , oral antibiotics , enzyme inducers , P-glycoprotein ( PgP ) inhibitors , cyclooxygenase-2 ( P35354 ) inhibitors , tumor necrosis factor-alpha ( P01375 ) inhibitors , or blockers of biliary excretion of SN-38 . Further studies are needed to identify the molecular targets associated with CPT-11 toxicity and safe and effective agents for alleviating CPT-11-induced diarrhea . Regulation of the natural killer cell response to interferon-alpha by biogenic amines . Monocytes , recovered from human peripheral blood by counter-current centrifugal elutriation ( CCE ) , suppressed baseline natural killer ( NK ) cell cytotoxicity ( NKCC ) and rendered NK cells resistant to activation of cytotoxicity by human recombinant interferon-alpha ( IFN-alpha ) by a cell contact-dependent mechanism . Monocyte-induced suppression of resting and IFN-activated NK cells was abrogated by the biogenic amines histamine [ via H2-type receptors ( P25021 ) ] and serotonin [ via P08908 -type receptors ( 5-HT1AR ) ] . Our data are suggestive of a monocyte/NK cell interaction that is subject to regulation by biogenic amines . [ Asthma with hypersensitivity to aspirin ] . DB00945 -induced asthma ( AIA ) is a distinct clinical entity characterised by hypersensitivity to aspirin and other non-steroidal anti-inflammatory drugs ( NSAIDs ) typically occurring in the form of asthma and severe rhinosinusitis . Its prevalence is about 10 % , but AIA probably is underdiagnosed . The diagnosis can only be established with certainty by challenge tests , using increasing doses of aspirin . The crucial cell in AIA is the eosinophil , while the role of mast cell is more elusive . At the biochemical level , AIA is distinguished by profound underlying arachidonic acid disturbances . An overproduction of leukotrienes was observed in patients with aspirin hypersensitivity at baseline and following aspirin challenge . In some patients altered prostaglandin release following aspirin was encountered . The inhibition of P23219 , but not P35354 , was shown to precipitate post-challenge symptoms precipitated by aspirin . The new insights into eicosanoid molecular biology and genetics have recently emerged . The patients with aspirin-sensitive asthma require often prolonged oral , inhaled and nasal corticotherapy . Antileukotrienes also can be used . DB00945 and NSAIDs should be avoided . Nevertheless , highly specific P35354 inhibitors are well tolerated . DB00945 desensitisation , followed by daily aspirin treatment , is a valuable therapeutic option . Green tea polyphenols in the prevention of colon cancer . Several plant-based nutrients and non-nutrients that can inhibit mutagenesis and proliferation have been identified . Some of the most promising nutrients identified as chemopreventive agents in colon cancer prevention include isoflavones , curcumin , calcium , vitamin D and more recently Green tea polyphenols ( GTP ) . In addition to inhibiting mutagenesis and proliferation , these compounds are relatively non-toxic , are of low cost and can be taken orally or as a part of the daily diet . Epidemiological and laboratory studies have identified epigallocatechin gallate ( EGCG ) in green tea polyphenols ( GTP ) , as the most potent chemopreventive agent that can induce apoptosis , suppress the formation and growth of human cancers including colorectal cancers ( CRC ) . It is only logical then , that future clinical studies should focus on examining the efficacy of phytochemicals such as EGCG in cancer chemoprevention as an alternative to pharmacological agents , especially in populations where administration of P35354 inhibitors , DB00945 and NSAIDS is contraindicated . The goal of this review is to provide the rationale , and discuss the use of EGCG in GTP as a chemopreventive agent for prevention of colon carcinogenesis and present evidence for the efficacy and safety of these agents based on epidemiological , animal , in vitro studies and Phase I clinical trials . Altered expression of beta-catenin , P12830 , cycloxygenase-2 , and p53 protein by ovine intestinal adenocarcinoma cells . Around 1.6 % of sheep in New Zealand develop small-intestinal adenocarcinomas . These neoplasms typically develop widespread metastases . The common development of these neoplasms and their subsequent behavior suggests that sheep could be a useful animal model of human colonic cancer . However , for an animal model of human disease to be relevant , similar genetic mutations should be present . Genetic mutations within human colonic cancers frequently result in expression of cycloxygenase-2 ( P35354 ) , loss of membranous expression of beta-catenin and P12830 , and accumulation of p53 protein within the neoplastic cells . Immunohistochemistry was used to investigate the presence of these 4 proteins within 26 ovine intestinal adenocarcinomas . Loss of membranous beta-catenin reactivity was observed in 14 of 26 ovine intestinal adenocarcinomas ( 54 % ) . The loss of membranous beta-catenin reactivity was accompanied by cytoplasmic and nuclear reactivity in 2 neoplasms . Loss of P12830 was observed within 8 of 26 neoplasms ( 31 % ) . Neoplastic cell expression of P35354 was observed in 12 of 26 neoplasms ( 46 % ) , whereas cells within 3 of 26 neoplasms ( 11 % ) contained visible p53 protein . In conclusion , all 4 proteins that commonly have altered expression in human colonic cancers were also altered in a proportion of the ovine intestinal adenocarcinomas . These results provide additional evidence that sheep could be useful for the study of human colonic cancer .

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P06401 -mediated up-regulation of transthyretin in preimplantation mouse uterus . P02766 ( P02766 ) , a carrier for thyroxine and retinol , has its messenger RNA ( mRNA ) expressed in the glandular endometrial epithelium and its protein detected in the glandular endometrial epithelium and the uterine lumen . P02766 mRNA is dramatically up-regulated in the preimplantation mouse uterus as well as the P-treated ovariectomized mouse uterus , and in both situations the up-regulation of P02766 is blocked by treatment with the P receptor antagonist DB00834 . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Recombinant human prothrombin kringle-2 induces bovine capillary endothelial cell cycle arrest at G0- P55008 phase through inhibition of cyclin D1/ P11802 complex : modulation of reactive oxygen species generation and up-regulation of cyclin-dependent kinase inhibitors . P00734 is a plasma glycoprotein involved in blood coagulation and , as we have previously reported , prothrombin kringles inhibit BCE ( bovine capillary endothelial ) cell proliferation . To reveal the mechanism , we investigated the influence of rk-2 ( recombinant human prothrombin kringle-2 ) on the BCE cell cycle progression and ROS ( reactive oxygen species ) generation using FACS ( fluorescence-activated cell sorter ) analysis . Cell cycle analysis showed a decrease of G(1) phase cells in cells treated with P09038 ( basic fibroblast growth factor ) and an increase in cells treated with rk-2 , as compared with the control cells . But , the portion of the S phase was reversed . In Western blot analysis , P09038 induced cytoplasmic translocation of P38936 (Waf1/Cip1) and p27(Kip1) and phosphorylation of p27(Kip1) but rk-2 treatment inhibited translocation of P38936 (Waf1/Cip1) and p27(Kip1) from nucleus to cytoplasm and phosphorylation of p27(Kip1) . Also , rk-2 induced up-regulation of p53 and nuclear P38936 (Waf1/Cip1) and inhibited the cyclin D1/ P11802 ( cyclin-dependent kinase 4 ) complex . The ROS level of rk-2-treated BCE cells was increased 2-fold when compared with the control , but treatment with Q9C000 ( N-Acetyl-L : -cysteine ) , an anti-oxidant , decreased ROS generation about 55 % as compared with the rk-2 treatment . Q9C000 treatment also restored cell cycle progression inhibited by rk-2 and down-regulated p53 and nuclear P38936 (Waf1/Cip1) expression induced by rk-2.These data suggest that rk-2 induces the BCE cell cycle arrest at G(0)-G(1) phase through inhibition of the cyclin D1/ P11802 complex caused by increase of ROS generation and nuclear cyclin-dependent kinase inhibitors . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Q99572 receptor-dependent intestinal afferent hypersensitivity in a mouse model of postinfectious irritable bowel syndrome . The DB00171 -gated P2X(7) receptor ( P2X(7)R ) was shown to be an important mediator of inflammation and inflammatory pain through its regulation of IL-1β processing and release . Trichinella spiralis-infected mice develop a postinflammatory visceral hypersensitivity that is reminiscent of the clinical features associated with postinfectious irritable bowel syndrome . In this study , we used P2X(7)R knockout mice ( P2X(7)R(-/-) ) to investigate the role of P2X(7)R activation in the in vivo production of IL-1β and the development of postinflammatory visceral hypersensitivity in the T. spiralis-infected mouse . During acute nematode infection , IL-1β-containing cells and P2X(7)R expression were increased in the jejunum of wild-type ( WT ) mice . Peritoneal and serum IL-1β levels were also increased , which was indicative of elevated IL-1β release . However , in the P2X(7)R(-/-) animals , we found that infection had no effect upon intracellular , plasma , or peritoneal IL-1β levels . Conversely , infection augmented peritoneal P01375 -α levels in both WT and P2X(7)R(-/-) animals . Infection was also associated with a P2X(7)R-dependent increase in extracellular peritoneal lactate dehydrogenase , and it triggered immunological changes in both strains . Jejunal afferent fiber mechanosensitivity was assessed in uninfected and postinfected WT and P2X(7)R(-/-) animals . Postinfected WT animals developed an augmented afferent fiber response to mechanical stimuli ; however , this did not develop in postinfected P2X(7)R(-/-) animals . Therefore , our results demonstrated that P2X(7)Rs play a pivotal role in intestinal inflammation and are a trigger for the development of visceral hypersensitivity . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Selective measurement of white matter and gray matter diffusion trace values in normal human brain . The trace of the diffusion tensor ( or simply the trace ) is diagnostically valuable for detecting acute ischemic lesions . A number of studies indicate that the trace of human gray matter ( GM ) and white matter ( WM ) are quite similar . This is somewhat surprising considering the different cellular environments of GM and WM . It is possible that partial volume averaging ( P32926 ) effects between GM and WM , inherent in many of the ultrafast imaging sequences used for diffusion measurements , are responsible for this observation . In order to minimize P32926 effects , the trace values of GM and WM have been selectively measured by implementing double inversion recovery ( P30518 ) echo planar imaging ( P10646 ) pulse sequences . Results on six normal volunteers indicate that the trace values of WM and GM are not statistically different . The ischemic preconditioning paradox in deceased donor liver transplantation-evidence from a prospective randomized single blind clinical trial . While animal studies show that ischemic preconditioning ( IPC ) is beneficial in liver transplantation ( LT ) , evidence from few smaller clinical trials is conflicting . From October 2003 to July 2006 , 101 deceased donors ( DD ) were randomized to 10 min IPC ( n = 50 ) or No IPC ( n = 51 ) . Primary objective was efficacy of IPC to decrease reperfusion ( RP ) injury . Both groups had similar donor risk index ( DRI ) ( 1.54 vs. 1.57 ) . Aminotransferases on days 1 and 2 were significantly greater ( p < 0.05 ) in IPC recipients . In multivariate analyses , IPC had an independent effect only on day 2 aspartate transferase . P00734 time , bilirubin and histological injury were similar in both groups . IPC had no significant effect on plasma P01375 , P05231 and P22301 in the donor and P01375 and P05231 in the recipient . In contrast , IPC recipients had a significant rise in systemic P22301 levels after RP ( p < 0.05 ) and had fewer moderate/severe rejections within 30 days ( p = 0.09 ) . Hospital stay was similar in both groups . One-year patient and graft survival in IPC versus No IPC were 88 % versus 78 % ( p = 0.1 ) and 86 versus 76 % ( p = 0.25 ) , respectively . IPC increases RP injury after DDLT , an ' IPC paradox ' . Other potential benefits of IPC are limited . IPC may be more effective in combination with other preconditioning regimens . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Effects of G- P04141 on systemic inflammation , coagulation and platelet activation in patients with acute myocardial infarction . INTRODUCTION : In the prospective , randomised , double-blind , placebo-controlled Regenerate Vital Myocardium by Vigorous Activation of Bone Marrow Stem Cells ( REVIVAL ) -2 trial patients with acute myocardial infarction ( AMI ) and successful mechanical reperfusion received granulocyte-colony stimulating factor ( DB00099 , 10 μg/kg KG s.c. ) or placebo for 5 days . Aim of this substudy was to assess the impact of G- P04141 on systemic inflammatory and procoagulant responses and platelet activation . METHODS AND RESULTS : Before and five days after DB00099 ( n=56 ) or placebo ( n=58 ) circulating cytokine concentrations of interleukin ( IL ) -1ß , P05231 , P10145 , P22301 , IL-12 and Tumor-Necrosis Factor-α ( P01375 -α were measured . P00734 fragment F1+2 and Tissue Factor activity served as a measure for activated coagulation . Platelet activation was characterized by cell surface expression of the activated fibrinogen receptor ( O95456 ) , P16109 and P29965 by flow cytometry . Administration of G- P04141 was associated with elevated P01375 -α and CRP concentrations compared to the placebo group after 5 days . Other cytokines ( IL-1ß , P05231 , P10145 , P22301 , IL-12 ) were comparable after treatment with G- P21583 or placebo . Similarly , circulating prothrombin fragments F1+2 , TF activity and platelet activation did not differ in both groups . CONCLUSION : Treatment with G- P04141 in patients with AMI was associated with enhanced proinflammatory P01375 -α and CRP levels but no activation of coagulation . Adenovirus-mediated local expression of human tissue factor pathway inhibitor eliminates shear stress-induced recurrent thrombosis in the injured carotid artery of the rabbit . The main cause of acute coronary syndrome may be recurrent thrombosis , which is initiated by the activation of the extrinsic coagulation pathway . P13726 ( TF ) pathway inhibitor ( P10646 ) efficiently inhibits an early step in this pathway by the formation of a complex with factor VIIa , TF , and factor Xa . We determined whether local P10646 gene transfer can inhibit thrombosis in an injured artery without inducing systemic side effects . Balloon-injured rabbit carotid arteries were infected with an adenoviral vector that expressed either human P10646 ( AdCATFPI ) or bacterial beta-galactosidase ( AdCALacZ ) . Two to 6 days after gene transfer , thrombosis was induced by the production of constant stenosis of the artery , and blood flow was measured continuously with an electromagnetic flow probe . A cyclic flow variation , which is thought to reflect the recurrent formation and dislodgment of mural thrombi , was observed in all AdCALacZ-infected arteries as well as in saline-infused arteries . In contrast , no cyclic flow variation was detectable in AdCATFPI-transfected arteries , even in the presence of epinephrine ( 1 microg. kg-1. min-1 infusion ) . P00734 time , activated partial thromboplastin time , and the ex vivo platelet aggregation induced by either adenosine diphosphate or collagen were unaltered in AdCATFPI-infected rabbits . We found that in vivo P10646 gene transfer into an injured artery completely inhibits the recurrent thrombosis induced by shear stress even in the presence of catecholamine , without affecting systemic coagulation status . Adenovirus-mediated local expression of P10646 may have the potential for the treatment of human thrombosis . P28702 polymorphisms do not explain functional differences in vitamins D and A response in Antineutrophil cytoplasmic antibody associated vasculitis patients . It has been suggested that the retinoid X receptor beta ( P28702 ) gene is a risk factor for Wegener 's granulomatosis . We addressed if there is a functional difference in the response to retinoic acid ( RA ) and vitamin D in Antineutrophil cytoplasmic antibody ( ANCA ) associated systemic vasculitis ( AASV ) patients and if this was associated with P28702 genotypes . TNFalpha and P22301 production were measured in whole blood assay from AASV patients ( n = 51 ) and healthy controls ( HC , n = 67 ) . One micromolar of 1,25-(OH)(2) D3 , 9-cis RA ( 9c-RA ) or all-trans RA ( DB00755 ) was added to the assay . Genotyping was performed for exons 7 and 2 of the P28702 gene and for a microsatellite in vicinity of the P28702 gene . Lipopolysaccharide ( LPS ) mediated TNFalpha production and P22301 were significantly lower in patients . Addition of 1,25-(OH)(2) D3 , DB00755 or 9c-RA , blunted TNFalpha production , more pronounced in patients . Although all three compounds inhibited P22301 production significantly in HC , only 1,25-(OH)(2) D3 was found to be effective in patients . Allele distribution of the P28702 microsatellite differed significantly between patients and HC . This was not found for the SNP in exons 2 and 7 . Genotype of the latter correlated with the ability of 1,25-(OH)(2) D3 and DB00755 to inhibit P22301 production . We provide immunological evidence for a functional difference in vitamins D and A responsiveness in AASV patients . Since the inhibition of TNFalpha was more effective in patients , vitamin D supplementation might be an additional therapeutical approach . Inhibition of prothrombin kringle-2-induced inflammation by minocycline protects dopaminergic neurons in the substantia nigra in vivo . P00734 kringle-2 ( pKr-2 ) , a domain of prothrombin , can cause the degeneration of mesencephalic dopaminergic neurons through microglial activation . However , the chemical products that inhibit pKr-2-induced inflammatory activities in the brain are still not well known . The present study investigated whether minocycline , a semisynthetic tetracycline derivative , could inhibit pKr-2-induced microglial activation and prevent the loss of nigral dopaminergic ( DA ) neurons in vivo . To address this question , rats were administered a unilateral injection of pKr-2 in the substantia nigra in the presence or absence of minocycline . Our results show that pKr-2 induces the production of proinflammatory cytokines , such as tumor necrosis factor-α ( P01375 -α ) and interleukin-1β ( IL-1β ) , and inducible nitric oxide synthase from the activated microglia . In parallel , 7 days after pKr-2 injection , tyrosine hydroxylase immunocytochemical analysis and western blot analysis showed a significant loss of nigral DA neurons . This neurotoxicity was antagonized by minocycline and the observed neuroprotective effects were associated with the ability of minocycline to suppress the expression of tumor necrosis factor-α , interleukin-1β , and nitric oxide synthase . These results suggest that minocycline may be promising as a potential therapeutic agent for the prevention of DA neuronal degeneration associated with pKr-2-induced microglial activation . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . P02766 as an index of liver function after acute paracetamol poisoning . Liver damage in a woman who had taken an overdose of paracetamol and dextropropoxyphene was assessed by monitoring serum prealbumin concentrations and by routine plasma enzyme determinations . The plasma aspartate aminotransferase returned to normal levels after 3 days , alkaline phosphatase was slow to show increases in activity , and serum albumin concentration was in the normal range throughout . P00734 -time , although initially very high , returned almost to normal as a result of the administration of plasma . In contrast , serum prealbumin concentration decreased significantly after 36 h and continued to decrease , showing the course of failing liver function , until the patient 's death 15 days after presentation . P02766 , a functional plasma protein synthesised in the liver , has a short half-life , is a true index of liver function , and seems to be a more reliable indicator of liver function in drug overdose than plasma enzymes , prothrombin-time , or plasma drug concentration . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . P50406 mediates defective brain development in monoamine oxidase A-deficient mouse embryos . Monoamine oxidases A and B ( P21397 and P27338 ) are enzymes of the outer mitochondrial membrane that metabolize biogenic amines . In the adult central nervous system , MAOs have important functions for neurotransmitter homeostasis . Expression of MAO isoforms has been detected in the developing embryo . However , suppression of P27338 does not induce developmental alterations . In contrast , targeted inhibition and knockdown of P21397 expression ( E7.5-E10.5 ) caused structural abnormalities in the brain . Here we explored the molecular mechanisms underlying defective brain development induced by P21397 knockdown during in vitro embryogenesis . The developmental alterations were paralleled by diminished apoptotic activity in the affected neuronal structures . Moreover , dysfunctional P21397 expression led to elevated levels of embryonic serotonin ( 5-hydroxytryptamine ( 5-HT ) ) , and we found that knockdown of serotonin receptor-6 ( 5-Htr6 ) expression or pharmacologic inhibition of 5-Htr6 activity rescued the P21397 knockdown phenotype and restored apoptotic activity in the developing brain . Our data suggest that excessive 5-Htr6 activation reduces activation of caspase-3 and -9 of the intrinsic apoptotic pathway and enhances expression of antiapoptotic proteins Bcl-2 and Bcl-XL . Moreover , we found that elevated 5-HT levels in P21397 knockdown embryos coincided with an enhanced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) and a reduction of proliferating cell numbers . In summary , our findings suggest that excessive 5-HT in P21397 -deficient mouse embryos triggers cellular signaling cascades via 5-Htr6 , which suppresses developmental apoptosis in the brain and thus induces developmental retardations . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis .

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interacts_with DB00398?

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A novel tescalcin-sodium/hydrogen exchange axis underlying sorafenib resistance in P36888 -ITD+ AML . Internal tandem duplication ( ITD ) of fms-like tyrosine kinase 3 ( P36888 ) in acute myeloid leukemia ( AML ) is associated with inferior clinical prognosis . DB00398 is effective in clearing leukemic blasts in chemorefractory P36888 -ITD(+) AML , but leukemia progression invariably occurs . Mechanisms of drug resistance are not completely understood . We hypothesized that a gene encoding tescalcin ( Q96BS2 ) , known to be upregulated at leukemia progression during continuous sorafenib treatment and activate an Na(+)/H(+) exchanger type-1 ( P19634 ) , may underlie tyrosine kinase inhibitor resistance . Q96BS2 was highly expressed in P36888 -ITD(+) AML lines MOLM-13 and MV4-11 , and its knockdown by small-interfering RNA lowered intracellular pH ( pHi ) and induced apoptosis . The results were recapitulated by treatment with an P19634 inhibitor , 5-(N,N-hexamethylene) amiloride ( HMA ) . Induction of sorafenib resistance in the MOLM-13 cell line ( M13-RE ) significantly increased its sensitivity to HMA . The later also enhanced suppression of P36888 signaling by sorafenib in otherwise resistant cell lines . HMA treatment of MOLM-13 and MV4-11 as well as primary P36888 -ITD(+) AML cells significantly reduced leukemia initiation in anti-CD122-primed NOD/SCID mouse xenotransplantation . These observations provided novel information about the pathogenetic role of a Q96BS2 - P19634 -pHi axis in mediating sorafenib resistance in AML . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Treatment of P36888 -ITD-positive acute myeloid leukemia relapsing after allogeneic stem cell transplantation with sorafenib . Patients with acute myeloid leukemia ( AML ) and internal tandem duplication of P07333 -like tyrosine kinase receptor-3 gene ( P36888 -ITD ) mutation have poor prognoses and are often treated with allogeneic hematopoietic stem cell transplantation ( HSCT ) . DB00398 , an inhibitor of multiple kinases including P36888 , has shown promising activity in P36888 -ITD-positive AML . We treated 16 patients with P36888 -ITD-positive AML who relapsed after HSCT with sorafenib alone ( n = 8 ) or in combination with cytotoxic chemotherapy ( n = 8 ) . The number of circulating blasts decreased in 80 % of cases , but none of the patients achieved complete remission ( CR ) ; 3 achieved partial remission . Two patients were bridged to a second transplantation but both relapsed within 3 months of the transplantation . Median overall survival ( OS ) was 83 days , with none surviving more than a year . DB00398 is not effective in the treatment of P36888 -ITD-positive AML relapsing after HSCT . Preventive strategies after HSCT may be more suitable for these high-risk patients . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . DB00398 ( BAY 43-9006 ) : review of clinical development . DB00398 ( BAY 43-9006 ) is a novel oral bis-aryl urea compound originally developed as an inhibitor to RAF kinase for its anti-proliferative property . It also inhibits receptor tyrosine kinases of multiple pro-angiogenic factors such as P35968 /3 , Flt-3/ and P09619 . The combination of both its anti-proliferative and anti-angiogenic properties makes sorafenib an attractive agent in cancer treatment . Phase I studies demonstrated that sorafenib was well tolerated , and the recommended phase II dose was 400 mg twice daily continuously . Common toxicities included skin toxicity ( rash and hand-foot syndrome ) , gastrointestinal toxicities ( nausea and diarrhea ) and fatigue . Anti-tumor activities were observed in multiple tumors types including renal cell carcinoma and hepatocellular carcinoma . Randomized phase III studies in these tumor types are ongoing , and results are eagerly waited . Involvement of protein kinase Cdelta/extracellular signal-regulated kinase/poly(ADP-ribose) polymerase-1 ( P09874 ) signaling pathway in histamine-induced up-regulation of histamine H1 receptor gene expression in HeLa cells . The histamine H(1) receptor ( P35367 ) gene is up-regulated in patients with allergic rhinitis . However , the mechanism and reason underlying this up-regulation are still unknown . Recently , we reported that the P35367 expression level is strongly correlated with the severity of allergic symptoms . Therefore , understanding the mechanism of this up-regulation will help to develop new anti-allergic drugs targeted for P35367 gene expression . Here we studied the molecular mechanism of P35367 up-regulation in HeLa cells that express P35367 endogenously in response to histamine and phorbol 12-myristate 13-acetate ( PMA ) . In HeLa cells , histamine stimulation caused up-regulation of P35367 gene expression . Rottlerin , a PKCδ-selective inhibitor , inhibited up-regulation of P35367 gene expression , but Go6976 , an inhibitor of Ca(2+)-dependent PKCs , did not . DB11320 or PMA stimulation resulted in PKCδ phosphorylation at DB00135 (311) and DB00156 (505) . Activation of PKCδ by H(2)O(2) resulted in P35367 mRNA up-regulation . Overexpression of PKCδ enhanced up-regulation of P35367 gene expression , and knockdown of the PKCδ gene suppressed this up-regulation . DB11320 or PMA caused translocation PKCδ from the cytosol to the Golgi . U0126 , an MEK inhibitor , and DPQ , a poly(ADP-ribose) polymerase-1 inhibitor , suppressed PMA-induced up-regulation of P35367 gene expression . These results were confirmed by a luciferase assay using the P35367 promoter . Phosphorylation of P29323 and P04049 in response to PMA was also observed . However , real-time PCR analysis showed no inhibition of P35367 mRNA up-regulation by a P04049 inhibitor . These results suggest the involvement of the PKCδ/ P29323 /poly(ADP-ribose) polymerase-1 signaling pathway in histamine- or PMA-induced up-regulation of P35367 gene expression in HeLa cells . Comparative transcriptional and functional profiling of clear cell and papillary renal cell carcinoma . Renal cell carcinoma ( RCC ) is known to effectively prevent immune recognition . However , little is known about the mechanisms that underlie this phenomenon . Thus , the identification of immunogenic molecules associated with RCC and the elucidation of the corresponding signaling pathways are crucial to the development of effective treatments . We performed transcriptional and functional profiling with cDNA microarrays ( 1070 cDNA probes ) on a total of 17 RCCs , 11 clear cell and 6 papillary , and on corresponding normal tissue . Samples were clustered based on their expression profiles . We found a total of 45 genes to be regulated equally by both tumor types compared to the normal tissue . A set of 13 differentially expressed genes was identified between the examined tumor subtypes . Functional analysis was performed for both gene sets and showed a significant enrichment of cell surface genes regulated in both tumor subtypes . Within these we found five surface marker genes to be upregulated ( O14763 , P32970 , P19438 , P09619 , and Q9Y275 ) which are involved in immune responses via the regulation of lymphocytes and can also induce apoptosis . Their overexpression in both tumor subtypes suggests a possible involvement in the immune escape strategies of RCC . The combination of transcriptional and functional profiling revealed potential target molecules for novel therapy strategies that must be studied in more detail . Combination of the P29323 inhibitor AZD6244 and low-dose sorafenib in a xenograft model of human renal cell carcinoma . DB00398 , a multikinase inhibitor , is currently used as monotherapy for advanced renal cell carcinoma ( RCC ) . However , adverse effects associated with its use have been experienced by some patients . In this study , we examined the antitumor and antiangiogenic activities of low-dose sorafenib in combination with the MEK inhibitor AZD6244 ( sorafenib/AZD6244 ) in a preclinical model of RCC . Primary RCC 08-0910 and RCC 786-0 cells as well as patient-derived RCC models were used to study the antitumor and antiangiogenic activities of sorafenib/AZD6244 . Changes of biomarkers relevant to angiogenesis and cell cycle were determined by western immunoblotting . Microvessel density , apoptosis and cell proliferation were analyzed by immunohistochemistry . Treatment of RCC 786-0 cells with sorafenib/AZD6244 resulted in P55008 cell cycle arrest and blockade of serum-induced cell migration . DB00398 /AZD6244 induced apoptosis in primary RCC 08-0910 cells at low concentrations . In vivo addition of AZD6244 to sorafenib significantly augmented the antitumor activity of sorafenib and allowed dose reduction of sorafenib without compromising its antitumor activity . DB00398 /AZD6244 potently inhibited angiogenesis and phosphorylation of P35968 , P09619 -β and P29323 , p90RSK , p70S6K , cdk-2 and retinoblastoma . DB00398 /AZD6244 also caused upregulation of p27 , Bad and Bim but downregulation of survivin and cyclin B1 . These resulted in a reduction in cellular proliferation and the induction of tumor cell apoptosis . Our findings showed that AZD6244 and sorafenib complement each other to inhibit tumor growth . This study provides sound evidence for the clinical investigation of low-dose sorafenib in combination with AZD6244 in patients with advanced RCC . DB00398 as treatment for relapsed or refractory pediatric acute myelogenous leukemia . The prognosis for children with acute myelogenous leukemia ( AML ) has improved with overall survival rates of up to 65 % [ Pui et al. J Clin Oncol 2011 ; 29 : 551-565 ] . However , the cure rate for AML lags behind that of acute lymphoblastic leukemia . Advances in AML leukemogenesis are leading to refined risk stratification . P07333 like tyrosine kinase 3 ( P36888 ) mutations are independently associated with a poor prognosis . Newer kinase inhibitors , including sorafenib , have shown promise in adult studies . We report three pediatric patients with relapsed AML who achieved a sustained remission with sorafenib . Further trials are necessary to understand the role of sorafenib in pediatric AML . B cell activating factor-dependent expression of vascular endothelial growth factor in MH7A human synoviocytes stimulated with tumor necrosis factor-α . Angiogenesis in rheumatoid arthritis ( RA ) is one of the histological hallmarks , which is mediated by expression of vascular endothelial growth factor ( P15692 ) in RA synovium . P15692 expression is enhanced by P01375 -α , the main pro-inflammatory cytokine in RA . B cell activating factor ( Q9Y275 ) which plays a role in maturation and maintenance of B cells is also associated with autoimmune RA . Here , we investigated whether Q9Y275 could regulate P15692 expression in P01375 -α-stimulated synovium using MH7A synovial cells that are established by transfection with the SV40 T antigen . Changes in hBAFF and hVEGF were measured by western blotting , RT-PCR and luciferase promoter assay . When MH7A cells were treated with P01375 -α , we observed that P01375 -α increased the expression of hBAFF and hVEGF . P01375 -α also increased transcriptional activity of hBAFF and hVEGF as judged by luciferase promoter assay . Inhibition of hBAFF expression with Q9Y275 -siRNA decreased transcriptional level and activity of hVEGF . In addition , when c-fos expression was inhibited by the transfection of MH7A cells with c-fos-siRNA , data showed that transcriptional level and activity of both hBAFF and hVEGF were attenuated by the activation with P01375 -α . Our results demonstrate for the first time that P15692 -mediated angiogenesis in RA could be controlled by P01375 -α-induced Q9Y275 expression through c-Fos . Data suggest that P01375 -α-induced Q9Y275 expression and Q9Y275 -mediated P15692 expression in synovium may cooperate to maintain the capacity of such cells to protect B cells from apoptosis and the supply of nutrients and oxygen in inflammatory microenvironments . A phase I trial of the oral , multikinase inhibitor sorafenib in combination with carboplatin and paclitaxel . PURPOSE : This study evaluated the safety , maximum tolerated dose , pharmacokinetics , and antitumor activity of sorafenib , a multikinase inhibitor , combined with paclitaxel and carboplatin in patients with solid tumors . PATIENTS AND METHODS : Thirty-nine patients with advanced cancer ( 24 with melanoma ) received oral sorafenib 100 , 200 , or 400 mg twice daily on days 2 to 19 of a 21-day cycle . All patients received carboplatin corresponding to AUC6 and 225 mg/m(2) paclitaxel on day 1 . Pharmacokinetic analyses were done for sorafenib on days 2 and 19 of cycle 1 and for paclitaxel on day 1 of cycles 1 and 2 . Pretreatment tumor samples from 17 melanoma patients were analyzed for P15056 mutations . RESULTS : DB00398 was well tolerated at the doses evaluated . The most frequent severe adverse events were hematologic toxicities ( grade 3 or 4 in 33 patients , 85 % ) . Twenty-seven ( 69 % ) patients had sorafenib-related adverse events , the most frequent of which were dermatologic events ( 26 patients , 67 % ) . Exposure to paclitaxel was not altered by intervening treatment with sorafenib . Treatment with sorafenib , paclitaxel , and carboplatin resulted in one complete response and nine partial responses , all among patients with melanoma . There was no correlation between P15056 mutational status and treatment responses in patients with melanoma . CONCLUSIONS : The recommended phase II doses are oral 400 mg twice daily sorafenib , carboplatin at an AUC6 dose , and 225 mg/m(2) paclitaxel . The tumor responses observed with this combined regimen in patients with melanoma warrant further investigation . Ferulic acid is nephrodamaging while gallic acid is renal protective in long term treatment of chronic kidney disease . BACKGROUNDS & AIMS : The long term therapeutic effect of ferulic acid ( FA ) and gallic acid ( GA ) in treatment of chronic kidney disease ( CKD ) has been lacking . METHODS : Doxorubicin ( DR , DB00997 ) -induced CKD rat model was established for this study . RESULTS : DR significantly reduced levels of serum albumin , GOT , GPT , RBC , P01375 -α , and urinary creatinine and elevated serum cholesterol , TG , BUN , creatinine , uric acid , WBC , platelet count , and P05231 . In DRCKD rats , FA and GA significantly increased kidney weight and glomerular volume . FA reduced glomerular filtration rate but GA did not . FA enhanced more collagen deposition than GA in renal cortex and glomeruli . Both FA and GA showed crucial hyperlipidemic activity . The inhibitory effects of FA and GA on P08253 were very comparable . GA suppressed P08253 more effectively than FA in DRCKD rats . Both FA and GA induced SOD elevation and MDA elimination . In DRCKD rats , Western blot analysis indicated that FA further up-regulated P28906 , α-SMA , tissue pDGFR , p- P09619 , and TGF-β ; and down-regulated p-PI3K , and p-Akt . Since both DB00102 and TGF-β are considered to induce kidney prefibrosis stage , GA was proved to be more beneficial in this regard . CONCLUSIONS : GA tends to protect the CKD while FA is not recommended for the long term CKD therapy . Endothelial progenitor cells in relation to endothelin-1 and endothelin receptor blockade : a randomized , controlled trial . AIMS : Endothelial progenitor cells ( EPC ) represent an endogenous repair mechanism involving rendothelialization and neoangiogenesis . Patients with both diabetes and vascular disease have low numbers of circulating EPC . The endothelium-derived peptide , endothelin-1 ( ET-1 ) , is increased in patients with type 2 diabetes and vascular complications and has been suggested to contribute to endothelial dysfunction . Therefore , we investigated the relation between EPC and plasma ET-1 and the effect of dual ET-1 receptor antagonist treatment . METHODS : In this double blind study patients with type 2 diabetes mellitus and microalbuminuria were randomized to treatment with the dual P25101 /ETB receptor antagonist DB00559 treatment ( 125mg bid ; n=17 ) or placebo ( n=19 ) for four weeks . Different EPC subpopulations were enumerated by flow cytometry using triple staining ( P28906 , CD133 , P35968 ) at baseline at the end of treatment . Viability was assessed by 7AAD and Annexin-V-staining . RESULTS : Baseline ET-1 levels correlated significantly with P02741 levels . Patients with ET-1 levels above the median value had higher levels of P28906 (+)CD133(+) and P28906 (+) P35968 (+) EPC . There was no difference in P28906 (+) and P28906 (+)CD133(+) P35968 (+) cells , markers of EPC apoptosis or circulating markers of endothelial damage between patients with ET-1 levels below or above the median . Four week treatment with DB00559 did not change EPC levels . CONCLUSION : Among patients with type 2 diabetes and vascular disease , high plasma levels of ET-1 are associated with higher number of EPC . The recruitment of EPC does not seem to be regulated via ET-1 receptor activation since treatment with a dual ET-1 receptor blocker did not affect circulating EPC numbers . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Role of the steroid receptor coactivator Q9Y6Q9 in cell growth . Steroid receptor coactivator 3 ( Q9Y6Q9 /p/CIP/ Q9Y6Q9 / Q9Y6Q9 /RAC3/ Q9Y6Q9 ) is a member of the P52701 family of nuclear receptor coactivators , which includes Q15788 ( Q15788 ) and P12931 -2 ( Q15596 /GRIP1/NCoA2 ) . Previous studies indicate that Q9Y6Q9 is required for normal animal growth and is often amplified or overexpressed in many cancers , including breast and prostate cancers . However , the mechanisms of Q9Y6Q9 -mediated growth regulation remain unclear . In this study , we show that overexpression of Q9Y6Q9 stimulates cell growth to increase cell size in prostate cancer cell lines . Furthermore , our results indicate that overexpression of Q9Y6Q9 can modulate the AKT signaling pathway in a steroid-independent manner , which results in the activation of AKT/ P42345 signaling concomitant with an increase in cell size . In contrast , down-regulation of Q9Y6Q9 expression in cells by small interfering RNA decreases cell growth , leading to a smaller cell size . Similarly , in Q9Y6Q9 null mutant mice , AKT signaling is down-regulated in normally Q9Y6Q9 -expressing tissues . Taken together , these results suggest that Q9Y6Q9 is an important modulator for mammalian cell growth . P01344 Producing Hepatocellular Carcinoma Treated with DB00398 : Metabolic Complications and a Foresight to Molecular Targeting Therapy to the IGF Signal . Hypoglycemia is a rare paraneoplastic manifestation of patients with neoplasms . Hypoglycemia can be induced by several causes , including an aberrant increase of hypoglycemic agents and adrenal insufficiency . DB00398 is the first agent to demonstrate a survival benefit in the treatment of advanced hepatocellular carcinoma ( HCC ) . This small molecule inhibits serine/threonine kinase RAF in tumor cells and tyrosine kinases VEGFR/ P09619 in tumor vasculature and decreases tumor growth and angiogenesis . In this paper , we report a case of HCC who was treated with sorafenib and showed severe hypoglycemia . This hypoglycemia might be induced by two causes , both adrenal insufficiency as an adverse effect of sorafenib and activation of the insulin-like growth factor ( IGF ) signal by excessive secretion of incompletely processed precursors of P01344 . Although the IGF signal is suggested to be involved in aberrant growth of HCC in some cases , there is no other report showing the influence of sorafenib on HCC with active IGF signal . Unfortunately , the effect of sorafenib was limited in the present case . However , emerging drugs that directly inhibit the IGF signal can be expected to be highly effective in the treatment of HCC with hypoglycemia . DB00398 monotherapy gives sustainable suppression of P36888 clone in untreated patients with P36888 -internal tandem duplication positive acute myeloid Leukaemia . DB00398 inhibits the angiogenesis and growth of orthotopic anaplastic thyroid carcinoma xenografts in nude mice . Anaplastic thyroid carcinoma ( ATC ) remains one of the most lethal human cancers . We hypothesized that sorafenib , a multikinase inhibitor of the BRaf , vascular endothelial growth factor receptor-2 , and platelet-derived growth factor receptor-beta kinase , would decrease tumor growth and angiogenesis in an orthotopic model of ATC . The in vitro antiproliferative and proapoptotic effects of sorafenib on ATC cell lines were examined . To study the in vivo effects of sorafenib on orthotopic ATC tumors in nude mice , sorafenib was given p.o. at 40 or 80 mg/kg daily . Intratumoral effects were studied using immunohistochemical analysis . The effect of sorafenib on survival of the mice was also studied . DB00398 inhibited the in vitro proliferation of ATC cell lines . DB00398 also significantly inhibited tumor angiogenesis via the induction of endothelial apoptosis in an orthotopic model of thyroid cancer . As result , the growth of orthotopic ATC xenografts was reduced and the survival of the test animals was improved . DB00398 exerts significant antitumor activity in an orthotopic xenograft model of ATC via a potent antiangiogenic effect . The antiangiogenic effects of sorafenib suggest that its use in clinical setting may not depend on the P15056 mutational status of thyroid tumors . Given the lack of curative options for patients with ATC , sorafenib warrants further study as a therapeutic agent against ATC . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Dependence on phosphoinositide 3-kinase and DB01367 -RAF pathways drive the activity of RAF265 , a novel RAF/ P35968 inhibitor , and RAD001 ( DB01590 ) in combination . Activation of phosphatidylinositol-3-kinase ( PI3K ) -AKT and Kirsten rat sarcoma viral oncogene homologue ( P01116 ) can induce cellular immortalization , proliferation , and resistance to anticancer therapeutics such as epidermal growth factor receptor inhibitors or chemotherapy . This study assessed the consequences of inhibiting these two pathways in tumor cells with activation of P01116 , PI3K-AKT , or both . We investigated whether the combination of a novel RAF/vascular endothelial growth factor receptor inhibitor , RAF265 , with a mammalian target of rapamycin ( P42345 ) inhibitor , RAD001 ( everolimus ) , could lead to enhanced antitumoral effects in vitro and in vivo . To address this question , we used cell lines with different status regarding P01116 , P42336 , and P15056 mutations , using immunoblotting to evaluate the inhibitors , and MTT and clonogenic assays for effects on cell viability and proliferation . Subcutaneous xenografts were used to assess the activity of the combination in vivo . RAD001 inhibited P42345 downstream signaling in all cell lines , whereas RAF265 inhibited RAF downstream signaling only in P15056 mutant cells . In vitro , addition of RAF265 to RAD001 led to decreased AKT , S6 , and P06730 binding protein 1 phosphorylation in HCT116 cells . In vitro and in vivo , RAD001 addition enhanced the antitumoral effect of RAF265 in HCT116 and H460 cells ( both P01116 mut , P42336 mut ) ; in contrast , the combination of RAF265 and RAD001 yielded no additional activity in A549 and MDAMB231 cells . The combination of RAF and P42345 inhibitors is effective for enhancing antitumoral effects in cells with deregulation of both DB01367 -RAF and PI3K , possibly through the cross-inhibition of 4E binding protein 1 and S6 protein . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . DB11320 contributes to tissue remodeling via periostin expression . DB11320 is thought to have a critical role in the synthesis of extracellular matrix in skin and may be involved in tissue remodeling of allergic diseases . Recent studies revealed that periostin , a matricelluar protein , contributed to tissue remodeling ; however , a link between periostin and histamine remains unproven . We investigated whether periostin was involved in histamine-induced collagen production . Cultured dermal fibroblasts derived from wild-type ( WT ) or periostin knockout ( PN(-/-) ) mice were stimulated with histamine , and then collagen and periostin production was evaluated . DB11320 induced collagen gene expression in WT fibroblasts in the late phase but not in the early phase , whereas no effect on collagen expression was observed in histamine-stimulated PN(-/-) fibroblasts . In WT fibroblasts , histamine directly induced periostin expression in a dose-dependent manner , and an H1 receptor antagonist blocked both periostin and collagen expression . DB11320 activated extracellular signal-regulated kinase 1/2 ( P27361 /2 ) through the H1 receptor . Q15063 induction was inhibited by either H1 antagonist or P27361 /2 inhibitor treatment in vitro and was attenuated in P35367 (-/-) mice . Elevated expression of periostin was found in lesional skin from atopic dermatitis patients . These results suggest that histamine mediates periostin induction and collagen production through activation of the H1 receptor-mediated P27361 /2 pathway ; furthermore , histamine may accelerate the chronicity of atopic dermatitis . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Synthesis , biological evaluation and 3D-QSAR study of novel 4,5-dihydro-1H-pyrazole thiazole derivatives as P15056 (V⁶⁰⁰E) inhibitors . Many reports implied that the P15056 serine/threonine kinase was mutated in various types of human tumors , which were related with cell growth , survival and differentiation . To provide new therapeutic opportunities , a series of novel 4,5-dihydro-1H-pyrazole derivatives ( 6a-10d ) containing thiazole moiety as potential V600E mutant P15056 kinase ( P15056 (V600E) ) inhibitors were designed and synthesized . All compounds were evaluated in vitro for anticancer activities against WM266.4 human melanoma cell line and breast cancer MCF-7 cell line . Compound 10d displayed the most potential antiproliferative activity with an IC50 value of 0.12μM against cell line WM266.4 and 0.16μM against MCF-7 with positive control DB00398 . Results of the inhibitory activity against P15056 (V600E) revealed that compound 10d was bearing the best bioactivity with IC50 of 0.05μM as well . On the basis of the result of flow cytometry , with the dose of compound 10d increasing , more and more cancer cell gradually encountered apoptosis or died , which indicated the compound 10d could induce remarkable apoptosis of MCF-7 and WM266.4 cells in a dose dependent manner . Furthermore , docking simulation of inhibitor analogues and 3D-QSAR modeling provided potential binding model and further knowledge of pharmacophore . DB00398 inhibits activation of human peripheral blood T cells by targeting P06239 phosphorylation . DB00398 , a novel drug for metastatic renal cancer , has broad-spectrum activity against multiple tyrosine kinases , including P04049 , vascular endothelial growth factor receptor and platelet-derived growth factor receptor . However , little is known about its effects on the immune system . In this report , we examine the effects of sorafenib on the proliferation and activation of human peripheral blood T cells , as well as its effects on T-cell-mediated immune response in mice . At concentrations similar to those used in patients , sorafenib inhibited the proliferation of primary human T cells in vitro . At more than 10 microM , sorafenib caused an irrecoverable inhibition of proliferation , even after drug withdrawal . In addition , sorafenib induced T-cell apoptosis at concentrations higher than 10 muM. sorafenib also caused G(0)/G(1) phase arrest , inhibition of CD25 and Q07108 expression , interleukin-2 production and P06239 phosphorylation in the T cells ; all of these effects exhibited dose and time dependence . When tested against contact dermatitis in mice , sorafenib significantly reduced the ear swelling induced by picryl chloride . These findings suggest that sorafenib may cause the loss of T-cell immune response by inducing apoptosis and targeting P06239 . This could potentially lead to immunosuppression in patients with cancer . Multi-kinase inhibition in ovarian cancer . DB00398 ( Nexavar ) is a multi-kinase inhibitor that was developed as an inhibitor of RAF-1 , in the P27361 /2 pathway , but which was subsequently shown to inhibit class III tyrosine kinase receptors . ( 1 ) More recently regorafenib ( Stivarga ) has been developed , which is a further fluorinated version of sorafenib with greater bioavailability and similar inhibitory properties against RAF-1/class III RTKs . ( 2 ) Some of the anti-tumor effects of sorafenib have been ascribed to anti-angiogenic actions of this agent on endothelial associated kinases such as P35968 . Other effects of sorafenib clearly have to be due to its effects on the inherent biology of the tumor cells themselves . For example , through various mechanisms sorafenib has been shown in the laboratory and the clinic to suppress expression of the protective protein Q8WXI8 -1 . ( 3 ) DB00398 has also been linked to inhibition of P40763 , NFκB , and activation of the death receptor CD95 . ( 4 ) DB00398 is routinely dosed daily ( 400 mg P55957 ) and 7 d after the start of dosing has a Cmax of ~21 μM with a nadir at 12 h of ~10 μM , and is a highly protein bound based on in vitro assays . ( 5 ) Despite this in vitro binding data sorafenib has profound in vivo effects on tumor cells in renal carcinoma and hepatocellular carcinoma patients ; cells which are not per se addicted to high activity oncogene signals that are targets of sorafenib/regorafenib . Thus the precise stable bioavailable level of sorafenib/regorafenib in patient plasma is not known . A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets . To establish the druggability of a target , genetic validation needs to be supplemented with pharmacological validation . Pharmacological studies , especially in the kinase field , are hampered by the fact that many reference inhibitors are not fully selective for one target . Fortunately , the initial trickle of selective inhibitors released in the public domain has steadily swelled into a stream . However , rationally picking the most selective tool compound out of the increasing amounts of available inhibitors has become progressively difficult due to the lack of accurate quantitative descriptors of drug selectivity . A recently published approach , termed ' selectivity entropy ' , is an improved way of expressing selectivity as a single-value parameter and enables rank ordering of inhibitors . We provide a guide to select the best tool compounds for pharmacological validation experiments of candidate drug targets using selectivity entropy . In addition , we recommend which inhibitors to use for studying the biology of the 20 most investigated kinases that are clinically relevant : Abl ( P00519 ) , P31749 , Q9UM73 , Aurora A/B , CDKs , MET , P07333 ( P07333 ) , P00533 , P36888 , P04626 ( P04626 ) , O14920 ( O14920 ) , O60674 /3 , P45983 /2/3 ( P45983 /9/10 ) , Q02750 /2 , P53350 , PI3Ks , p38α ( Q16539 ) , P15056 , P12931 and P35968 ( P35968 ) . Initial testing ( stage 1 ) of the multi-targeted kinase inhibitor sorafenib by the pediatric preclinical testing program . BACKGROUND : DB00398 is an inhibitor of multiple kinases ( e.g. , P15692 receptors , P09619 , P36888 , P07949 , P15056 , P10721 ) and is approved by FDA for treatment of two adult cancers . The activity of sorafenib was evaluated against the PPTP 's in vitro and in vivo panels . PROCEDURES : DB00398 was evaluated against the PPTP in vitro panel using 96-hr exposure at concentrations ranging from 1.0 nM to 10.0 µM . It was tested against the PPTP in vivo panels at a dose of 60 mg/kg administered by oral gavage daily for 5 days per week , repeated for 6 weeks . RESULTS : In vitro sorafenib demonstrated cytotoxic activity , with a median IC(50) value of 4.3 µM . Twenty of 23 cell lines had IC(50) values between 1.0 and 10.0 µM . A single cell line ( Kasumi-1 ) with an activating P10721 mutation had an IC(50) value < 1.0 µM ( IC(50) = 0.02 µM ) . In vivo sorafenib induced significant differences in event-free survival ( O43281 ) distribution compared to control in 27 of 36 ( 75 % ) of the evaluable solid tumor xenografts and in 1 of 8 ( 12.5 % ) of the evaluable ALL xenografts . DB00398 induced tumor growth inhibition meeting criteria for intermediate activity ( O43281 T/C ) in 15 of 34 ( 44 % ) evaluable solid tumor xenografts . No xenografts achieved an objective response . CONCLUSIONS : The primary in vitro activity of sorafenib was noted at concentrations above 1 µM , with the exception of a more sensitive cell line with an activating P10721 mutation . The primary in vivo effect for sorafenib was tumor growth inhibition , which was observed across multiple histotypes . Role of sorafenib in the treatment of advanced hepatocellular carcinoma : An update . DB00398 is the first and only p.o. administrated drug currently approved to treat advanced hepatocellular carcinoma ( HCC ) . However , concerns have been raised about sorafenib therapy , including acquired drug resistance . This review provides an overview of sorafenib in the treatment of HCC on the basis of data obtained in the laboratory and in clinical studies . Three underlying mechanisms have been found to support sorafenib therapy . First , sorafenib blocks HCC cell proliferation by inhibiting BRaf and Raf1/c-Raf serine/threonine kinase phosphorylation in the mitogen-activated protein kinase pathway . Second , sorafenib induces apoptosis by reducing elF4E phosphorylation and downregulating Mcl-1 levels in tumor cells . Third , sorafenib prevents tumor-associated angiogenesis by inactivating vascular endothelial growth factor receptors ( P35968 and -3 ) and the platelet-derived growth factor receptor-β . Clinical trials have demonstrated the effectiveness and relative safety of sorafenib , and thus the drug is used in unresectable HCC . However , many patients may develop acquired resistance to sorafenib , so their response to sorafenib is eventually lost . DB00398 may induce autophagy , which leads to apoptosis . However , autophagy can also cause drug resistance . Many studies have combined sorafenib with other treatments in an effort to increase its effects , reduce the necessary dose or overcome resistance . It is urgent to study the mechanisms underlying how sorafenib interacts with cellular molecules and other drugs to increase its efficacy and reduce resistance in HCC patients . Synergistic cytotoxicity and molecular interaction on drug targets of sorafenib and gemcitabine in human pancreas cancer cells . BACKGROUND : Current treatments have a modest impact on survival of pancreatic cancer patients and this study investigates the interaction between sorafenib and gemcitabine and the molecular pharmacodynamics of this combination . METHODS : The pancreatic cancer cells were treated with sorafenib and gemcitabine , alone or in combination . The effects of treatments were evaluated on cell proliferation , cell cycle , apoptosis , phosphorylation of Akt , c-Kit , P29323 and P35968 , and expression of genes related to drug activity . RESULTS : Gemcitabine and sorafenib synergistically interacted on the inhibition of cell proliferation , and assessment of apoptosis demonstrated that drug associations increased the apoptotic index . DB00398 reduced c-Kit , P29323 and P35968 activation and on the other hand , gemcitabine inhibited Akt phosphorylation . Moreover , quantitative PCR showed that sorafenib modulated the expression of genes related to gemcitabine activity , while gemcitabine induced the expression of P30086 . CONCLUSION : These data demonstrate that gemcitabine and sorafenib combination displays a synergistic effect in pancreatic cancer cells . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of P35968 /Flk-1 . Previous studies revealed that gambogic acid ( GA ) , the major active ingredient of gamboge , a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia , possessed significant anticancer activity both in vitro and in vivo . In this study , we explored the high antiangiogenic activities of GA for the first time . GA inhibits the P15692 -stimulated proliferation , migration and tube formation of human umbilical vein endothelial cells ( HUVECs ) as well as microvessel sprouting from rat aortic rings in vitro . Moreover , GA inhibits vessel growth in matrigel plugs and P62158 in vivo and transplanted tumor in mice . The results also indicated that GA decreases P15692 production of cultured tumor cells and inhibits P15692 -induced tyrosine phosphorylation of P35968 /Flk-1 . This inhibition of receptor phosphorylation is correlated with a significant decrease in P15692 -triggered phosphorylated forms of P29323 , AKT and p38 . Taken together , these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor . Treatment of hepatocellular carcinoma with adenoviral vector-mediated P49771 gene therapy . P36888 ligand ( P49771 ) plays an important role in development and activation of dendritic cells ( DCs ) and natural killer cells ( NK ) . It has been shown that administration of either tumor cells transfected in vitro with P49771 vectors or soluble P49771 fusion protein in a high dose can enhance host antitumor immunity in animal model systems . In this study , we developed a recombinant defective adenovirus with an insert of gene encoding extracellular domain of mouse P49771 ( Ad-mFlt3L ) under control of cytomegalovirus promoter and investigated its biological efficacy in eliciting tumor-specific immune response against hepatocellular carcinoma in mouse hepatoma model . The constructed Ad-mFlt3L efficiently infected hepa 1-6 hepatoma cells both in vitro and in vivo , leading to a high production of mFlt3L proteins in association with accumulation of DCsNK cells and lymphocytes in local tumor tissues . Tumor cells infected with Ad-mFlt3L lost tumorigenicity and became more immunogenic in syngeniec animal models . Intratumoral injection of Ad-mFlt3L ( 10(9) expression-forming unit ) x 3 significantly inhibited tumor growth with elicitation of long-lasting antitumor immunity , which is both preventive and curative . The tumor-specific immunity can be partially abrogated by depletion of either CD3+ P01730 + T cells or NK cells and can be also re-established in naïve animals by adoptive transfer of splenocytes from treated mice . The results suggest that adenovirus-mediated P49771 gene therapy may provide a useful strategy for treatment of cancers . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . A randomized phase I clinical and biologic study of two schedules of sorafenib in patients with myelodysplastic syndrome or acute myeloid leukemia : a NCIC ( National Cancer Institute of Canada ) Clinical Trials Group Study . DB00398 is a small molecule inhibitor of RAF kinase , P35968 , c- P10721 , and P36888 . In this randomized phase I study , eligible patients had relapsed/refractory acute myeloid leukemia ( AML ) , and one prior induction regimen , or were age > 65 with untreated myelodysplastic syndrome ( P43034 ) or secondary AML . DB00398 was given orally for 28 days ( cont ) or 14 days ( int ) every 4 weeks at three dose levels ( 100 , 200 , and 400 mg P55957 ) ; 300 mg cont was also tested . Forty-two patients were enrolled ( median age 71 [ 37-82 ] ; prior chemotherapy : 22 ) . Dose-limiting toxicity ( DLT ) was : 100 mg P55957 : 0/7 patients ; 200 mg P55957 : 2/12 patients ; 400 mg P55957 : 1/17 patients . DB00398 400 mg cont was not tolerated in this population : 6/8 received < 14 days of treatment due to toxicity ; no DLT was seen with 300 mg cont . One CR was seen in a patient with AML with P36888 -ITD . Flow cytometry studies suggest that sorafenib inhibits P29323 phosphorylation via c- P10721 . The recommended phase II dose in AML is 300 mg P55957 continuously , and testing in combination and in P36888 -ITD AML is warranted . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Pharmacologic Inhibition of MNKs in Acute Myeloid Leukemia . The Ras/Raf/MAPK and PI3K/Akt/ P42345 pathways are key signaling cascades involved in the regulation of cell proliferation and survival , and have been implicated in the pathogenesis of several types of cancers , including acute myeloid leukemia ( AML ) . The oncogenic activity of P06730 driven by the Mnk kinases is a convergent determinant of the two cascades , suggesting that targeting the Mnk/ P06730 axis may provide therapeutic opportunity for the treatment of cancer . Herein , a potent and selective Q9HBH9 inhibitor ( MNKI-85 ) and a dual-specific Q9BUB5 and Q9HBH9 inhibitor ( MNKI-19 ) , both derived from a thienopyrimidinyl chemotype , were selected to explore their antileukemic properties . MNKI-19 and MNKI-85 are effective in inhibiting the growth of AML cells that possess an M5 subtype with P36888 -internal tandem duplication mutation . Further mechanistic studies show that the downstream effects with respect to the selective Q9BUB5 /2 kinase inhibition in AML cells causes P55008 cell cycle arrest followed by induction of apoptosis . MNKI-19 and MNKI-85 demonstrate similar Q9HBH9 kinase activity and cellular antiproliferative activity but exhibit different time-dependent effects on cell cycle progression and apoptosis . Collectively , this study shows that pharmacologic inhibition of both Q9BUB5 and Q9HBH9 can result in a more pronounced cellular response than targeting Q9HBH9 alone . However , MNKI-85 , a first-in-class inhibitor of Q9HBH9 , can be used as a powerful pharmacologic tool in studying the Q9HBH9 / P06730 -mediated tumorigenic mechanism . In conclusion , this study provides a better understanding of the mechanism underlying the inhibition of AML cell growth by Mnk inhibitors and suggests their potential utility as a therapeutic agent for AML . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Exploring schizophrenia drug-gene interactions through molecular network and pathway modeling . In this study , we retrieved 39 schizophrenia-related antipsychotic drugs from the DrugBank database . These drugs had interactions with 142 targets , whose corresponding genes were defined as drug targeted genes . To explore the complexity between these drugs and their related genes in schizophrenia , we constructed a drug-target gene network . These genes were overrepresented in several pathways including : neuroactive ligand-receptor pathways , glutamate metabolism , and glycine metabolism . Through integrating the pathway information into a drug-gene network , we revealed a few bridge genes connected the sub-networks of the drug-gene network : Q12879 , O60391 , Q14957 , Q13224 , P21728 , and P14416 . These genes encode ionotropic glutamate receptors belonging to the DB01221 receptor family and dopamine receptors . DB00502 was the only drug to directly interact with these pathways and receptors and consequently may have a unique action at the drug-gene interaction level during the treatment of schizophrenia . This study represents the first systematic investigation of drug-gene interactions in psychosis . Investigation of immunomodulatory properties of human Wharton 's Jelly-derived DB05914 after lentiviral transduction . Human Wharton 's Jelly-derived Mesenchymal Stem Cells ( hWJ-MSCs ) are considered as an alternative for bone-marrow-derived MSCs . These cells have immunosuppressive properties . It was unclear whether the WJ-MSCs would sustain their immunomodulatory characteristics after lentiviral transduction or not . In this study , we evaluated immunomodulatory properties of WJ-MSCs after lentiviral transduction . HWJ-MSCs were transduced with lentiviral particles . Expression of transduced and un-transduced hWJ-MSCs surface molecules and secretion of P22301 , P14210 , P15692 and TGF-β was analyzed . Cell proliferation and frequency of P01730 (+)CD25(+) CD127(low/neg) Foxp3(+) T regulatory cells was measured . There was no difference between the surface markers and secretion of P22301 , P14210 , P15692 and TGF-β in transduced and un-transduced hWJ-MSCs . Both cells inhibited the proliferation of PHA stimulated PBMCs , and improved the frequency of T regulatory cells . These findings suggest that lentiviral transduction does not alter the immunomodulatory function of hWJ-MSCs . However , lentiviral transduction may have a wide range of applications in gene therapy . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . AP-1 transrepressing retinoic acid does not deplete coactivators or AP-1 monomers but may target specific Jun or Fos containing dimers . Retinoic acid ( RA ) inhibits tumor promotion in many models in vivo and in vitro , among them mouse epidermal JB6 cells . RA treatment suppresses 12-O-tetradecanoylphorbol-13-acetate ( TPA ) induced AP-1 activity , an activity that is required for transformation of JB6 P+ cells . The molecular mechanism of AP-1 transrepression by retinoids is unclear , especially as related to inhibition of transformation . Overexpression of AP-1 components did not rescue TPA induced AP-1 activation nor did a Q86UG4 pull down experiment implicate direct binding , thus rendering unlikely both a Jun/Fos-RA-RAR direct interaction and a Jun/Fos sequestration mechanism . Overexpression of p300 , Q15788 or pCAF did not abrogate AP-1 suppression by RA , thus arguing against coactivator competition . Overexpression of the corepressor silencing mediator for retinoic acid and thyroid hormone receptors ( Q9Y618 ) suppressed AP-1 activity . However , Q9Y618 but not RA inhibited cJun transactivation , suggesting Q9Y618 does not mediate RA transrepression . RA treatment also did not block TPA induced P29323 phosphorylation , Jun/Fos family protein expression except for cFos , or DNA binding of the AP-1 complex . The transcriptional activities of full-length JunB and full-length Fra-1 , but not the transactivation domain fusions , were increased by TPA treatment and suppressed by RA . Since these full-length fusions have bzip domains , the results suggest that JunB and/or Fra-1-containing dimers may constitute one target of RA for transrepression of AP-1 . DB01590 ( RAD001 ) : an P42345 inhibitor for the treatment of metastatic renal cell carcinoma . The recent introduction of drugs that inhibit angiogenesis or the P42345 has provided new options for the treatment of metastatic renal cell carcinoma , a disease which often has a poor prognosis . Chemotherapy and cytokine therapy are largely ineffective . The 5-year survival rate is under 10 % . DB01590 , an immunosuppressive drug widely used for the prevention of allograft rejection and an P42345 inhibitor , is one of the latest drugs undergoing clinical trials in metastatic renal cell carcinoma . It has been tested in patients with progressive disease after therapy with tyrosine kinase receptor inhibitors ( sunitinib , sorafenib or both ) , which interfere with signaling pathways , such as the P15692 pathway . Clinical efficacy results ( progression-free survival ) for everolimus are promising and the safety profile is good . The ability of sorafenib to inhibit oncogenic PDGFRbeta and P36888 mutants and overcome resistance to other small molecule inhibitors . BACKGROUND AND OBJECTIVES : Activated tyrosine kinases are implicated in the pathogenesis of chronic and acute leukemia , and represent attractive targets for therapy . DB00398 ( BAY43-9006 , Nexavar ) is a small molecule B-RAF inhibitor that is used for the treatment of renal cell carcinoma , and has been shown to have activity against receptor tyrosine kinases from the platelet-derived growth factor receptor ( P09619 ) and vascular endothelial growth factor receptor ( VEGFR ) families . We investigated the efficacy of sorafenib at inhibiting mutants of the receptor tyrosine kinases PDGFRbeta , P10721 , and P36888 , which are implicated in the pathogenesis of myeloid malignancies . DESIGN AND METHODS : We tested the effect of sorafenib on the proliferation of hematopoietic cells transformed by P41212 -PDGFRbeta , P36888 with an internal tandem duplication or D835Y point mutation , and the P10721 (D816V) mutant . The direct effect of sorafenib on the activity of these kinases and their downstream signaling was tested using phospho-specific antibodies . RESULTS : We show that sorafenib is a potent inhibitor of P41212 -PDGFRbeta and P36888 mutants , including some of the mutants that confer resistance to PKC412 and other P36888 inhibitors . DB00398 induced a cell cycle block and apoptosis in the acute myeloid leukemia cell lines MV4-11 and MOLM-13 , both expressing P36888 with an internal tandem duplication , whereas no effect was observed on four other acute myeloid leukemia cell lines . The imatinib-resistant P10721 (D816V) mutant , associated with systemic mastocytosis , was found to be resistant to sorafenib . INTERPRETATION AND CONCLUSIONS : These results warrant further clinical studies of sorafenib for the treatment of myeloid malignancies expressing activated forms of PDGFRbeta and P36888 . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . DB00398 priming may augment salvage chemotherapy in relapsed and refractory P36888 -ITD-positive acute myeloid leukemia . DB00398 in patients with advanced biliary tract carcinoma : a phase II trial . BACKGROUND : Advanced biliary tract carcinoma has a very poor prognosis , with chemotherapy being the mainstay of treatment . DB00398 , a multikinase inhibitor of P35968 /-3 , P09619 , B-Raf , and C-Raf , has shown to be active in preclinical models of cholangiocarcinoma . METHODS : We conducted a phase II trial of single-agent sorafenib in patients with advanced biliary tract carcinoma . DB00398 was administered at a dose of 400 mg twice a day . The primary end point was the disease control rate at 12 weeks . RESULTS : A total of 46 patients were treated . In all , 26 ( 56 % ) had received chemotherapy earlier , and 36 patients completed at least 45 days of treatment . In intention-to-treat analysis , the objective response was 2 % and the disease control rate at 12 weeks was 32.6 % . Progression-free survival ( PFS ) was 2.3 months ( range : 0-12 months ) , and the median overall survival was 4.4 months ( range : 0-22 months ) . Performance status was significantly related to PFS : median PFS values for ECOG 0 and 1 were 5.7 and 2.1 months , respectively ( P=0.0002 ) . The most common toxicities were skin rash ( 35 % ) and fatigue ( 33 % ) , requiring a dose reduction in 22 % of patients . CONCLUSIONS : DB00398 as a single agent has a low activity in cholangiocarcinoma . Patients having a good performance status have a better PFS . The toxicity profile is manageable . Cribriform-morular variant of papillary thyroid carcinoma in an 8-year-old girl : a case report with immunohistochemical and molecular testing . The description of the histological features and the immunohistochemical and molecular analyses of a case of cribriform-morular variant of papillary thyroid carcinoma in an 8-year-old girl with a family history of adenomatous polyposis is presented . The neoplasm was multifocal and bilateral , with a mixed pattern of solid , trabecular , and morular areas . The neoplasm showed angioinvasive behavior , extracapsular infiltration with extension to the perythyroidal muscles , and lymph node metastases . Tumor cells were positive for P62158 5.2 , cytokeratins 5/6 , Q15669 -1 , HBME-1 , galectin-3 , and β-catenin . In addition , the molecular tests did not reveal P15056 mutations , P07949 /PTC rearrangement , P25054 mutation , or P01116 mutation . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . Off-label use of cetuximab plus sorafenib and panitumumab plus regorafenib to personalize therapy for a patient with V600E P15056 -mutant metastatic colon cancer . DB00398 , the first agent developed to target P15056 mutant melanoma , is a multi-kinase inhibitor that was approved by the FDA for therapy of kidney and subsequently liver cancer , and is currently in clinical trials for thyroid , lung and brain cancer . Colorectal cancer with V600E P15056 mutation has shown relative resistance to standard chemotherapy regimens , as well as lack of efficacy to vemurafenib in clinical trials . New treatments are needed for P15056 -mutant colorectal cancer . We report a case of a patient with P15056 -mutant metastatic colon cancer whose disease had progressed on FOLFOX plus bevacizumab and subsequent FOLFIRI plus cetuximab . Based on preclinical data published in Nature in 2012 suggesting that successful therapeutic targeting of P15056 in colorectal cancer may require concomitant targeting of the P00533 , we offered this patient without other attractive options the combination of sorafenib plus cetuximab , in off-label use with informed consent . DB00398 and cetuximab therapy led to a mixed radiographic response with some areas showing dramatic improvement and other areas showing stable disease over a 7-month period which is a notably long period of progression-free survival for V600E P15056 mutated colon cancer . The cetuximab plus sorafenib therapy was very well-tolerated by the patient who remained on it long enough until another therapy option , regorafenib , was approved in September 2012 . The patient was offered single agent regorafenib at the time of progression . At the time of progression on single agent regorafenib , panitumumab was combined with regorafenib and this was also well-tolerated and appeared to slow disease progression . Further study of these approaches in the clinic as personalized treatment of P15056 -mutant advanced colorectal cancer is warranted . Acyclic retinoid inhibits angiogenesis by suppressing the MAPK pathway . Acyclic retinoid ( P10323 ) is currently under clinical trial as an agent to suppress the recurrence of hepatocellular carcinoma ( HCC ) through its ability to induce apoptosis in premature HCC cells . P10323 has an anticancer effect in vivo as well , although it shows weak apoptosis-inducing activity against mature HCC cells , suggesting the existence of an additional action mechanism . In this study , we investigated the antiangiogenic activity of P10323 . P10323 inhibited angiogenesis within chicken chorioallantoic membrane ( P62158 ) in as similar a manner as all-trans retinoic acid ( atRA ) . Although suppression of angiogenesis by atRA was partially rescued by the simultaneous addition of angiopoietin-1 , suppression of angiogenesis by P10323 was not rescued under the same condition at all . Conversely , although suppression of angiogenesis by P10323 was partially inverted by the simultaneous addition of vascular endothelial growth factor ( P15692 ) , suppression of angiogenesis by atRA was not affected under the same condition . These results suggested that mechanisms underlying the suppression of angiogenesis by P10323 and atRA were different . P10323 selectively inhibited the phosphorylation of P15692 receptor 2 ( P35968 ) and of extracellular signal-regulated kinase ( P29323 ) without changing their protein expression levels , and inhibited endothelial cell growth , migration , and tube formation . The inhibition of the phosphorylation of P29323 , endothelial growth , migration , tube formation , and angiogenesis by P10323 was rescued by the overexpression of constitutively active mitogen-activated protein kinase ( MAPK ) . Finally , P10323 , but not atRA , inhibited HCC-induced angiogenesis in a xenografted P62158 model . These results delineate the novel activity of P10323 as an antiangiogenic through a strong inhibition of the P35968 MAPK pathway . Evidence That Q12879 Mutations in Melanoma Correlate with Decreased Survival . Previous whole-exome sequencing has demonstrated that melanoma tumors harbor mutations in the Q12879 gene . Q12879 encodes the regulatory Q12879 subunit of the glutamate-gated N-methyl-d-aspartate receptor ( NMDAR ) , involvement of which in melanoma remains undefined . Here , we sequenced coding exons of Q12879 in 19 low-passage melanoma cell lines derived from patients with metastatic melanoma . Potential mutation impact was evaluated in silico , including within the Q12879 crystal structure , and clinical correlations were sought . We found that of 19 metastatic melanoma tumors , four ( 21 % ) carried five missense mutations in the evolutionarily conserved domains of Q12879 ; two were previously reported . Melanoma cells that carried these mutations were treatment-naïve . Sorting intolerant from tolerant analysis predicted that S349F , G762E , and P1132L would disrupt protein function . When modeled into the crystal structure of Q12879 , G762E was seen to potentially alter Q05586 - Q12879 interactions and ligand binding , implying disruption to NMDAR functionality . Patients whose tumors carried non-synonymous Q12879 mutations had faster disease progression and shorter overall survival ( P < 0.05 ) . This was in contrast to the P15056 V600E mutation , found in 58 % of tumors but showing no correlation with clinical outcome ( P = 0.963 ) . Although numbers of patients in this study are small , and firm conclusions about the association between Q12879 mutations and poor clinical outcome can not be drawn , our results highlight the high prevalence of Q12879 mutations in metastatic melanoma and suggest for the first time that mutated NMDARs impact melanoma progression . Complementary and alternative medicine use and quality of life in patients with primary brain tumors . This study explored the use of complementary and alternative medicine ( P62158 ) approaches and their relationship with demographic and disease characteristics and quality of life ( QOL ) in the primary brain tumor ( P10721 ) population . One hundred one P10721 patients were enrolled in this study . The results showed that 34 % of patients reported using P62158 . Forty-one percent reported using more than one type of P62158 . The average cost of each P62158 used per month was 69 dollars , with 20 % of patients spending more than 100 dollars per month . The majority ( 74 % ) reported that their physicians were unaware of their use of P62158 . Data analysis found a higher performance status to be the only factor significantly related to use of P62158 therapy ( P < 0.005 ) . There was no difference in patient report of QOL between users and nonusers of P62158 therapies . The high number of patients who do not report P62158 use has potential implications for evaluation of symptoms and response to therapy in this population . This may be especially relevant in those patients with higher functional status participating in clinical trials . Targeting PIM kinases impairs survival of hematopoietic cells transformed by kinase inhibitor-sensitive and kinase inhibitor-resistant forms of P36888 and P11274 / P00519 . Previous studies have shown that activation of the signal transducer and activator of transcription 5 ( P42229 ) plays an essential role in leukemogenesis mediated through constitutive activated protein tyrosine kinases ( PTK ) . Because PIM-1 is a P42229 target gene , we analyzed the role of the family of PIM serine/threonine kinases ( PIM-1 to PIM-3 ) in PTK-mediated transformation of hematopoietic cells . Ba/ P13726 cells transformed to growth factor independence by various oncogenic PTKs ( P41212 / O60674 , P41212 / Q16288 , P41212 / P00519 , P11274 / P00519 , P36888 -ITD , and H4/PDGFbetaR ) show abundant expression of PIM-1 and PIM-2 . Suppression of PIM-1 activity had a negligible effect on transformation . In contrast , expression of kinase-dead PIM-2 mutant ( PIM-2KD ) led to a rapid decline of survival in Ba/ P13726 cells transformed by P36888 -ITD but not by other oncogenic PTKs tested . Coexpression of PIM-1KD and PIM-2KD abrogated growth factor-independent growth of Ba/ P13726 transformed by several PTKs , including P11274 / P00519 . Targeted down-regulation of PIM-2 by RNA interference ( RNAi ) selectively abrogated survival of Ba/ P13726 cells transformed by various P36888 ( P36888 ) -activating mutants [ internal tandem duplication ( ITD ) and kinase domain ] and attenuated growth of human cell lines containing P36888 mutations . Interestingly , cells transformed by P36888 and P11274 / P00519 mutations that confer resistance to small-molecule tyrosine kinase inhibitors were still sensitive to knockdown of PIM-2 , or PIM-1 and PIM-2 by RNAi . Our observations indicate that combined inactivation of PIM-1 and PIM-2 interferes with oncogenic PTKs and suggest that PIMs are alternative therapeutic targets in PTK-mediated leukemia . Targeting the PIM kinase family could provide a new avenue to overcome resistance against small-molecule tyrosine kinase inhibitors . DB00398 enhances pemetrexed cytotoxicity through an autophagy-dependent mechanism in cancer cells . DB00642 ( ALIMTA ) is a folate anti-metabolite that has been approved for the treatment of non-small cell lung cancer , and has been shown to stimulate autophagy . In the present study , we sought to further understand the role of autophagy in the response to pemetrexed and to test if combination therapy could enhance the level of toxicity through altered autophagy in tumor cells . The multikinase inhibitor sorafenib ( NEXAVAR ) , used in the treatment of renal and hepatocellular carcinoma , suppresses tumor angiogenesis and promotes autophagy in tumor cells . We found that sorafenib interacted in a greater than additive fashion with pemetrexed to increase autophagy and to kill a diverse array of tumor cell types . Tumor cell types that displayed high levels of cell killing after combination treatment showed elevated levels of AKT , P08133 S6K and/or phosphorylated P42345 , in addition to class III RTKs such as PDGFRb and P17948 , known in vivo targets of sorafenib . In xenograft and in syngeneic animal models of mammary carcinoma and glioblastoma , the combination of sorafenib and pemetrexed suppressed tumor growth without deleterious effects on normal tissues or animal body mass . Taken together , the data suggest that premexetred and sorafenib act synergistically to enhance tumor killing via the promotion of a toxic form of autophagy that leads to activation of the intrinsic apoptosis pathway , and predict that combination treatment represents a future therapeutic option in the treatment of solid tumors . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Renal cell carcinoma and the use of sorafenib . Immunotherapy results in a small overall survival advantage in metastatic renal cell carcinoma ( RCC ) , but there is a need to develop more effective systemic therapies . Angiogenesis has an important role in the pathophysiology of RCC and vascular endothelial growth factor ( P15692 ) is a key mediator of this process . DB00398 ( BAY 43-9006 ) is a new agent belonging to a class of drugs called kinase inhibitors and inhibits the P15692 , platelet-derived growth factor ( PDGF ) , and c- P10721 receptor tyrosine kinases , amongst others . DB00398 has shown significant activity with manageable toxicity in metastatic RCC in phase 2 studies in patients pretreated with immunotherapy , whilst prolonged progression-free survival in comparison with placebo in a phase 3 study has been reported . Further phase 3 trials in advanced disease are ongoing and a trial of adjuvant sorafenib therapy in RCC is planned . DB00398 does not improve efficacy of chemotherapy in advanced pancreatic cancer : A GISCAD randomized phase II study . BACKGROUND : The RAF-MEK- P29323 pathway is commonly activated in pancreatic cancer because of a high frequency of P01116 - P15056 mutations . A phase II randomized trial was designed to investigate the activity of sorafenib in combination with chemotherapy in advanced pancreatic cancer . METHODS : Locally advanced or metastatic pancreatic adenocarcinoma patients were randomized in a 1:1 ratio to receive cisplatin plus gemcitabine with sorafenib 400mg bid ( arm A ) or without sorafenib ( arm B ) . RESULTS : One hundred and fourteen patients were enrolled ; of these , 43 ( 74.6 % ) patients progressed in arm A and 44 ( 82.4 % ) in arm B . Median progression-free survival was 4.3 months ( 95 % CI : 2.7-6.5 ) and 4.5 months ( 95 % CI : 2.5-5.2 ) , respectively ( HR=0.92 ; 95 % CI : 0.62-1.35 ) . Median overall survival was 7.5 ( 95 % CI : 5.6-9.7 ) and 8.3 months ( 95 % CI : 6.2-8.7 ) , respectively ( HR=0.95 ; 95 % CI : 0.62-1.48 ) . Response rates were 3.4 % in arm A and 3.6 % in arm B . CONCLUSIONS : DB00398 does not significantly enhance activity of chemotherapy in advanced pancreatic cancer patients , and therefore should not be assessed in phase III trials . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . [ DB00398 (Nexavar) ] . DB00398 (Nexavar)is a multikinase inhibitor , with disruptive activity at intracellular C-RAF , B-RAF and mutant P15056 receptors , and extracellular C- P10721 , P36888 , P35968 , P35916 and PDGFRb receptors . In the phase III study , as compared with placebo , treatment with sorafenib significantly prolonged progression free survival(PFS)in patients with advanced renal cell carcinoma in whom previous therapy has failed . Diarrhea , rash , fatigue , hand-foot skin reactions , and hypertension were the most common adverse events associated with sorafenib . As sorafenib was associated with similar rates of clinically manageable side effects in elderly patients as compared to younger patients , response rates to sorafenib in elderly patients were comparable to those of younger patients . DB00398 was approved multinationally for the treatment of advanced and/or metastatic renal cell carcinoma . DB00398 and sunitinib are reference standards of care for the treatment of advanced renal cell carcinoma and are recommended by current clinical guidelines . For the future , research of biomarker , adverse drug reaction , and combined regimens are needed to maximize the effects of molecular-targeted drugs .

MH_train_1044

MH_train_1044

MH_train_1044

interacts_with DB00188?

multiple_choice

Experimental autoimmune encephalomyelitis in the Wistar rat : dependence of MBP-specific T cell responsiveness on P33681 costimulation . Experimental autoimmune encephalomyelitis ( EAE ) is an animal model of human multiple sclerosis that requires the activation of autoreactive T cells for the expression of pathology . EAE has been most frequently studied in the Lewis rat model as well as in several murine models of EAE including the PLJ and B10PL strains . In the present study we describe a novel model of EAE induced in the Wistar rat strain by immunization with guinea pig spinal cord antigens and pertussis toxin ( PT ) . T cell responses were induced to myelin basic protein . Autoreactive T cells could be totally blocked by the in vitro treatment with DB01281 , a protein that blocks the costimulation of autoreactive T cells . The addition of P60568 could reverse the inhibition seen in vitro with DB01281 . The effects of inhibition of P33681 costimulation were also examined by an analysis of cytokine responses and P60568 receptor on T cells . DB01281 treatment in vitro reduced the expression of P60568 receptor on T cells , enhanced T cell apoptosis and decreased the synthesis of P60568 , P01579 and P01375 . DB01281 treatment had no effect on P22301 synthesis by T cells , a cytokine implicated in the functions of regulatory T cell subsets . Overall , our studies support the rationale of P33681 blocking therapies as a potential treatment for models of multiple sclerosis . The induction of EAE in the Wistar rat provides yet another novel model in which to examine the regulation of T cell autoimmunity . Knockdown of human deubiquitinase O00487 induces cell cycle arrest and senescence . The O00487 ( O00487 , also known as Rpn11/MPR1/S13/CepP1 ) protein within the 19S complex ( 19S cap ; PA700 ) is responsible for substrate deubiquitination during proteasomal degradation . The role of O00487 in cell proliferation and senescence was explored using siRNA knockdown in carcinoma cell lines . Our results reveal that down-regulation of O00487 by siRNA transfection had a considerable impact on cell viability causing cell arrest in the G0- P55008 phase , ultimately leading to senescence . The molecular events associated with decreased cell proliferation , cell cycle arrest and senescence include down-regulation of cyclin B1- P06493 - P30307 , down-regulation of cyclin D1 and up-regulation of P38936 (/Cip) and p27(/Kip1) . Most notably , phosphorylation of the retinoblastoma protein was markedly reduced in O00487 knockdown cells . A comparative study with P28074 , a subunit of the 20S proteasome , revealed that P28074 and O00487 have different effects on cell cycle , senescence and associated molecular events . These data support the view that the 19S and 20S subunits of the proteasome have distinct biological functions and imply that targeting 19S and 20S would have distinct molecular consequences on tumor cells . G(alpha)12/13 inhibition enhances the anticancer effect of bortezomib through P28074 downregulation . DB00188 is a proteasome inhibitor approved for anticancer therapy . However , variable sensitivity of tumor cells exists in this therapy probably due to differences in the expression of proteasome subunits . G(alpha)(12/13) serves modulators or signal transducers in diverse pathways . This study investigated whether cancer cells display differential sensitivity to bortezomib with reference to G(alpha)(12/13) expression , and if so , whether G(alpha)(12/13) affects the expression of proteasome subunits and their activities . DB00188 treatment exhibited greater sensitivities in Huh7 and SNU886 cells ( epithelial type ) than SK-Hep1 and SNU449 cells ( mesenchymal type ) that exhibited higher levels of G(alpha)(12/13) . Overexpression of an active mutant of G(alpha)(12) ( Galpha(12)QL ) or G(alpha)(13) ( G(alpha)(13)QL ) diminished the ability of bortezomib to induce cytotoxicity in Huh7 cells . Moreover , transfection with the minigene that disturbs G protein-coupled receptor-G protein coupling ( CT12 or Q9NXZ2 ) increased it in SK-Hep1 cells . Consistently , MiaPaCa2 cells transfected with CT12 or Q9NXZ2 exhibited a greater sensitivity to bortezomib . Evidence of G(alpha)(12/13) 's antagonism on the anticancer effect of bortezomib was verified in the reversal by G(alpha)(12)QL or G(alpha)(13)QL of the minigenes ' enhancement of cytotoxity . Real-time polymerase chain reaction assay enabled us to identify P28074 , multicatalytic endopeptidase complex-like-1 , and proteasome activator subunit-1 repression by CT12 or Q9NXZ2 . Furthermore , G(alpha)(12/13) inhibition enhanced the ability of bortezomib to repress P28074 , as shown by immunoblotting and proteasome activity assay . Moreover , this inhibitory effect on P28074 was attenuated by G(alpha)G(alpha)(12)QL or G(alpha)(13)QL . In conclusion , the inhibition of G(alpha)(12/13) activities may enhance the anticancer effect of bortezomib through P28074 repression , providing insight into the G(alpha)(12/13) pathway for the regulation of proteasomal activity . Regulation of P28074 protein and β subunits of mammalian proteasome by constitutively activated signal transducer and activator of transcription 3 ( P40763 ) : potential role in bortezomib-mediated anticancer therapy . The ubiquitin-proteasome system facilitates the degradation of ubiquitin-tagged proteins and performs a regulatory role in cells . Elevated proteasome activity and subunit expression are found in several cancers . However , the inherent molecular mechanisms responsible for increased proteasome function in cancers remain unclear despite the well investigated and defined role of the mammalian proteasome . This study was initiated to elucidate the mechanisms involved in the regulation of β subunits of the mammalian proteasome . Suppression of P40763 tyrosine phosphorylation coordinately decreased the mRNA and protein levels of the β subunits of the 20 S core complex in DU145 cells . Notably , P28074 , a molecular target of bortezomib , was shown to be a target of P40763 . Knockdown of P40763 decreased P28074 protein . Inhibition of phospho- P40763 substantially reduced P28074 protein levels in cells expressing constitutively active- P40763 . Accumulation of activated P40763 resulted in the induction of P28074 promoter and protein levels . In addition , a direct correlation was observed between the endogenous levels of P28074 and constitutively active P40763 . P28074 and P40763 protein levels remained unaltered following the inhibition of proteasome activity . The P01133 -induced concerted increase of β subunits was blocked by inhibition of the P01133 receptor or P40763 but not by the PI3K/AKT or MEK/ P29323 pathways . Decreased proteasome activities were due to reduced protein levels of catalytic subunits of the proteasome in P40763 -inhibited cells . Combined treatments with bortezomib and inhibitor of P40763 abrogated proteasome activity and enhanced cellular apoptosis . Overall , we demonstrate that aberrant activation of P40763 regulates the expression of β subunits , in particular P28074 , and the catalytic activity of the proteasome . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Hypomethylating agents reactivate O43524 in acute myeloid leukemia . The deregulation of the DNA damage response ( DDR ) can contribute to leukemogenesis and favor the progression from myelodysplastic syndrome ( P43034 ) to acute myeloid leukemia ( AML ) . Since hypomethylating agent , notably azacitidine , constitute an efficient therapy for patients with high-risk P43034 , we assessed whether such compounds can activate the DDR in malignant blasts . While azacitidine and decitabine had moderate effects on apoptosis and cell cycle progression , both agents induced profound changes in the expression and functionality of DDR-related proteins . Decitabine , and to a lesser degree azacitidine , induced the activation of checkpoint kinases Chk-1 and Chk-2 and the phosphorylation of the DDR-sensor P16104 . In addition , hypomethylating agents were found to cause the dephosphorylation of the transcriptional regulator forkhead box O3 , best known as O43524 , whose phosphorylation has been related to poor prognosis in AML . The dephoasphorylation of O43524 induced by azacitidine or decitabine in malignant blasts was accompanied by the translocation of O43524 from the cytoplasm to the nucleus . Upon stimulation with azacitidine , P43034 /AML-derived , azacitidine-sensitive SKM-1S cells upregulated O43524 and the pro-apoptotic O43524 targets O43521 and PUMA , and this effect was attenuated or abolished in azacitidine-resistant SMK-1R cells . Altogether , our results suggest that the reactivation of O43524 may contribute to the effects of hypomethylating agents in malignant blasts . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Proteasomal serine hydrolases are up-regulated by and required for influenza virus infection . Interactions between viruses and their host cells are important determinants of virus replication and of immune responses to the virus . However , these interactions and resulting consequences of these interactions remain poorly defined . Numerous recent quantitative proteomic approaches have measured host proteins affected by virus infection . Here , we used activity-based protein profiling ( P05067 ) to measure functional alterations in host serine hydrolases after influenza A virus infection of Madin-Darby canine kidney and human A549 lung cells . We identified 62 serine proteases . We then combined the P05067 approach with stable isotope labeling to directly measure how serine hydrolase activities were affected by virus infection . Differentially regulated SHs mapped into a few key cellular pathway systems , most notably the proteasomal system . The specific serine protease inhibitors DB06692 and Pefablock and specific proteasomal inhibitors DB00188 and MG132 significantly inhibited influenza virus growth . Some inhibitors also down-regulated activities of several proteasomal proteins , including P25786 , P25787 , and PMSB3 . Genetic knockdown of PMSA2 also attenuated influenza virus replication . These findings further our understanding of enzymatic cellular processes affected by influenza virus and may be beneficial in the search for additional antiviral therapeutic targets . Cytotoxic effects of bortezomib in myelodysplastic syndrome/acute myeloid leukemia depend on autophagy-mediated lysosomal degradation of Q9Y4K3 and repression of P25786 . DB00188 ( Velcade ) is used widely for the treatment of various human cancers ; however , its mechanisms of action are not fully understood , particularly in myeloid malignancies . DB00188 is a selective and reversible inhibitor of the proteasome . Paradoxically , we find that bortezomib induces proteasome-independent degradation of the Q9Y4K3 protein , but not mRNA , in myelodysplastic syndrome ( P43034 ) and acute myeloid leukemia ( AML ) cell lines and primary cells . The reduction in Q9Y4K3 protein coincides with bortezomib-induced autophagy , and subsequently with apoptosis in P43034 /AML cells . RNAi-mediated knockdown of Q9Y4K3 sensitized bortezomib-sensitive and -resistant cell lines , underscoring the importance of Q9Y4K3 in bortezomib-induced cytotoxicity . DB00188 -resistant cells expressing an shRNA targeting Q9Y4K3 were resensitized to the cytotoxic effects of bortezomib due to down-regulation of the proteasomal subunit α-1 ( P25786 ) . To determine the molecular consequences of loss of Q9Y4K3 in P43034 /AML cells , in the present study , we applied gene-expression profiling and identified an apoptosis gene signature . Knockdown of Q9Y4K3 in P43034 /AML cell lines or patient samples resulted in rapid apoptosis and impaired malignant hematopoietic stem/progenitor function . In summary , we describe herein novel mechanisms by which Q9Y4K3 is regulated through bortezomib/autophagy-mediated degradation and by which it alters P43034 /AML sensitivity to bortezomib by controlling P25786 expression . Subclones with the t(9;22)/ P11274 - P00519 rearrangement occur in AML and seem to cooperate with distinct genetic alterations . In AML , cooperation of mutations suppressing differentiation ( ' class-II-mutations ' ) with ' class-I-mutations ' increasing cell proliferation is frequent . In rare cases of myeloid malignancies , the P11274 - P00519 fusion was reported to cooperate as class-I-mutation with class-II-mutations , but most cases had to be classified as blast phase of chronic myeloid leukaemia ( CML ) . We identified five cases of Philadelphia positive subclones in AML occurring in coincidence with other genetic lesions : 1:220 patients with inv(16)/ Q13951 - P35749 ( 0·5 % ) , 2:272 AML cases with t(8;21)/ Q01196 - Q06455 ( 0·7 % ) , 1:1029 P06748 -mutated AML ( 0·1 % ) , and one patient with s-AML following P43034 with a 5q-deletion . Four patients had m- P11274 ( e1a2 ) P11274 - P00519 transcripts ; one case only had an M- P11274 ( b3a2 ) breakpoint . These cases allow some interesting conclusions : The P11274 - P00519 rearrangement apparently can cooperate with the P06748 mutation similar to other class-I-mutations . The identification of Philadelphia positive subclones in < 1 % of patients with Q03701 -leukaemias fits well with previous observations that most Q03701 -AML are accompanied by activating mutations in genes enhancing proliferation . Since we observed the occurrence of the Philadelphia positive subclones at diagnosis , at relapse , or throughout the disease , the time point of the emergence of Philadelphia subclones seems variable in AML . Clinical research should further concentrate on Philadelphia positive subclones in AML to assess the clinical impact . Analysis of a 26-kb region linked to the Mhc in zebrafish : genomic organization of the proteasome component beta/transporter associated with antigen processing-2 gene cluster and identification of five new proteasome beta subunit genes . Sequencing of zebrafish ( Danio rerio ) bacterial artificial chromosome and P1 artificial chromosome genomic clone fragments and of cDNA clones has led to the identification of five new loci coding for beta subunits of proteasomes ( PSMB ) . Together with the four genes identified previously , nine PSMB genes have now been defined in the zebrafish . Six of the nine genes reside in the zebrafish MHC ( Mhc ) class I region , four of them reside in a single cluster closely associated with TAP2 on a 26-kb long genomic fragment , and two reside at some distance from the fragment . In addition to homologues of the human genes P28074 through P28065 , two new genes , A5LHX3 and PSMB12 , have been found for which there are no known corresponding genes in humans . The new genes reside in the PSMB cluster in the Mhc . Homology and promoter region analysis suggest that the Mhc-associated genes might be inducible by P01579 . The zebrafish class I region contains representatives of three phylogenetically distinguishable groups of PSMB genes , X , Y , and Z . It is proposed that these genes were present in the ancestral PSMB region before Mhc class I genes became associated with it . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Pin1 promotes degradation of Smad proteins and their interaction with phosphorylated tau in Alzheimer 's disease . AIMS : Neurodegeneration in Alzheimer 's disease ( AD ) is characterized by pathological protein aggregates and inadequate activation of cell cycle regulating proteins . Recently , Smad proteins were identified to control the expression of AD relevant proteins such as P05067 , P11802 and CDK inhibitors , both critical regulators of cell cycle activation . This might indicate a central role for Smads in AD pathology where they show a substantial deficiency and disturbed subcellular distribution in neurones . Still , the mechanisms driving relocation and decrease of neuronal Smad in AD are not well understood . However , Pin1 , a peptidyl-prolyl-cis/trans-isomerase , which allows isomerization of tau protein , was recently identified also controlling the fate of Smads . Here we analyse a possible role of Pin1 for Smad disturbances in AD . METHODS : Multiple immunofluorescence labelling and confocal laser-scanning microscopy were performed to examine the localization of Smad and Pin1 in human control and AD hippocampi . Ectopic Pin1 expression in neuronal cell cultures combined with Western blot analysis and immunoprecipitation allowed studying Smad level and subcellular distribution . Luciferase reporter assays , electromobility shift , RNAi-technique and qRT-PCR revealed a potential transcriptional impact of Smad on Pin1 promoter . RESULTS : We report on a colocalization of phosphorylated Smad in AD with Pin1 . Pin1 does not only affect Smad phosphorylation and stability but also regulates subcellular localization of Q15796 and supports its binding to phosphorylated tau protein . Smads , in turn , exert a negative feed-back regulation on Pin1 . CONCLUSION : Our data suggest both Smad proteins and Pin1 to be elements of a vicious circle with potential pathogenetic significance in AD . P40763 restrains Q9Y6Q6 - and O00206 -mediated signalling by suppressing expression of the E2 ubiquitin-conjugating enzyme P61088 . The transcriptional regulator P40763 curbs pro-inflammatory cytokine production mediated by NF-κB signalling in innate immune cells , yet the mechanism by which this occurs has been unclear . Here we identify P40763 as a pivotal negative regulator of P61088 , an E2 ubiquitin-conjugating enzyme that facilitates Q9Y4K3 K63-linked ubiquitination and NF-κB activation . P61088 accumulates intracellularly in the absence of P40763 . Depletion of P61088 in Stat3-deficient macrophages subdues excessive O14788 - or LPS-dependent gene expression , indicating that P61088 overexpression mediates enhanced transcriptional responses in the absence of P40763 . In O14788 -activated macrophages , P40763 is stimulated by autocrine P05231 and inhibits accrual of Ets-1 , Set1 methyltransferase and trimethylation of histone H3 lysine 4 ( H3K4me3 ) at the Ube2n ( P61088 ) promoter . These results delineate a mechanism by which P40763 operates as a transcriptional repressor on Ube2n , thus modulating NF-κB activity by regulation of P61088 abundance . Our data suggest that this pathway plays important roles in bone homeostasis and restraint of inflammation . Partial least squares based gene expression analysis in estrogen receptor positive and negative breast tumors . BACKGROUND : Breast cancer is categorized into two broad groups : estrogen receptor positive ( ER+ ) and ER negative ( ER- ) groups . Previous study proposed that under trastuzumab-based neoadjuvant chemotherapy , tumor initiating cell ( Q8NDX1 ) featured ER- tumors response better than ER+ tumors . Exploration of the molecular difference of these two groups may help developing new therapeutic strategies , especially for ER- patients . MATERIALS AND METHODS : With gene expression profile from the Gene Expression Omnibus ( GEO ) database , we performed partial least squares ( PLS ) based analysis , which is more sensitive than common variance/regression analysis . RESULTS : We acquired 512 differentially expressed genes . Four pathways were found to be enriched with differentially expressed genes , involving immune system , metabolism and genetic information processing process . Network analysis identified five hub genes with degrees higher than 10 , including P05067 , P03372 , P84022 , Q92769 , and Q13131 . CONCLUSIONS : Our findings provide new understanding for the molecular difference between Q8NDX1 featured ER- and ER+ breast tumors with the hope offer supports for therapeutic studies . Chromatin acetylation , β-amyloid precursor protein and its binding partner O00213 in DNA double strand break repair . Among post-translational modifications of chromatin proteins taking place in DNA double strand break ( DSB ) repair , acetylation plays a prominent role . This review lists several facts and hypotheses concerning this process . Lack of acetyltransferase Q92993 ( HIV-Tat interacting protein of 60 kDa ) activity results in cells with defective DSB repair . The enzyme is present in the nucleus in a multimeric protein complex . Q92993 dependent activation of Q13315 ( ataxia telangiectasia mutated kinase ) is an early event in the response to DNA breakage . Other important acetylations are those of histones H4 and γ P16104 . Correct reconstruction of the damaged site is critical for survival and prevention of genetic and epigenetic changes in the cell that may affect the function of its daughter cells . Recently , two proteins with previously unsuspected functions in DSB repair have been identified as active in this process : Alzheimer β-amyloid precursor protein ( P05067 ) and its binding partner O00213 , β-amyloid precursor binding protein . Their participation in DSB repair in both neuronal and non-neuronal cells is related to acetylation carried out by the acetyltransferase complex . The same function is ascribed to heterochromatin protein 1 ( P59665 ) . So far , the relations ( if any ) between Q92993 activation by P59665 and by the O00213 complex remain unidentified . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . DB00188 -resistant myeloma cell lines : a role for mutated P28074 in preventing the accumulation of unfolded proteins and fatal ER stress . DB00188 is an effective agent for treating multiple myeloma ( MM ) . To investigate the underlying mechanisms associated with acquired resistance to this agent , we established two bortezomib-resistant MM cell lines , KMS-11/BTZ and OPM-2/BTZ , the 50 % inhibitory concentration values of which were respectively 24.7- and 16.6-fold higher than their parental cell lines . No activation of caspase and BH3-only proteins such as Noxa was noted in bortezomib-resistant cells after exposure to the drug . The accumulation of polyubiquitinated proteins was reduced in bortezomib-resistant cells compared with the parental cells , associated with avoidance of catastrophic ER stress as assessed by downregulation of P35638 expression . These resistant MM cells have a unique point mutation , G322A , in the gene encoding the proteasome beta5 subunit ( P28074 ) , likely resulting in conformational changes to the bortezomib-binding pocket of this subunit . KMS-11 parental cells transfected to express mutated P28074 also showed reduced bortezomib-induced apoptosis compared with those expressing wild-type P28074 or the parental cells . Expression of mutated P28074 was associated with the prevention of the accumulation of unfolded proteins . Thus , a fraction of MM cells may acquire bortezomib resistance by suppressing apoptotic signals through the inhibition of unfolded protein accumulation and subsequent excessive ER stress by a mutation of the P28074 gene . Ameliorating replicative senescence of human bone marrow stromal cells by P28074 overexpression . Multipotent human bone marrow stromal cells ( hBMSCs ) potentially serve as a source for cell-based therapy in regenerative medicine . However , in vitro expansion was inescapably accompanied with cell senescence , characterized by inhibited proliferation and compromised pluripotency . We have previously demonstrated that this aging process is closely associated with reduced 20S proteasomal activity , with down-regulation of rate-limiting catalytic β-subunits particularly evident . In the present study , we confirmed that proteasomal activity directly contributes to senescence of hBMSCs , which could be reversed by overexpression of the β5-subunit ( P28074 ) . Knocking down P28074 led to decreased proteasomal activity concurrent with reduced cell proliferation in early-stage hBMSCs , which is similar to the senescent phenotype observed in late-stage cells . In contrast , overexpressing P28074 in late-stage cells efficiently restored the normal activity of 20S proteasomes and promoted cell growth , possibly via upregulating the P12004 D1/ P11802 complex . Additionally , P28074 could enhance cell resistance to oxidative stress , as evidenced by the increased cell survival upon exposing senescent hBMSCs to hydrogen peroxide . Furthermore , P28074 overexpression retained the pluripotency of late-stage hBMSCs by facilitating their neural differentiation both in vitro and in vivo . Collectively , our work reveals a critical role of P28074 in 20S proteasome-mediated protection against replicative senescence , pointing to a possible strategy for maintaining the integrity of culture-expanded hBMSCs by manipulating the expression of P28074 . No evidence of mutations of the P28074 ( beta-5 subunit of proteasome ) in a case of myeloma with clinical resistance to DB00188 . Discovery and optimization of sulfonyl acrylonitriles as selective , covalent inhibitors of protein phosphatase methylesterase-1 . The serine hydrolase protein phosphatase methylesterase-1 ( Q9Y570 ) regulates the methylesterification state of protein phosphatase 2A ( PP2A ) and has been implicated in cancer and Alzheimer 's disease . We recently reported a fluorescence polarization-activity-based protein profiling ( fluopol- P05067 ) high-throughput screen for Q9Y570 that uncovered a remarkably potent and selective class of aza-β-lactam ( P00519 ) Q9Y570 inhibitors . Here , we describe a distinct set of sulfonyl acrylonitrile inhibitors that also emerged from this screen . The optimized compound , 28 ( AMZ30 ) , selectively inactivates Q9Y570 and reduces the demethylated form of PP2A in living cells . Considering that 28 is structurally unrelated to P00519 inhibitors of Q9Y570 , these agents , together , provide a valuable set of pharmacological probes to study the role of methylation in regulating PP2A function . We furthermore observed that several serine hydrolases were sensitive to analogues of 28 , suggesting that more extensive structural exploration of the sulfonyl acrylonitrile chemotype may result in useful inhibitors for other members of this large enzyme class . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . DB00072 has preferential activity against breast cancers driven by P04626 homodimers . In breast cancer cells with P04626 gene amplification , P04626 receptors exist on the cell surface as monomers , homodimers , and heterodimers with P00533 / P21860 . The therapeutic antibody trastuzumab , an approved therapy for P04626 (+) breast cancer , can not block ligand-induced P04626 heterodimers , suggesting it can not effectively inhibit P04626 signaling . Hence , P04626 oligomeric states may predict the odds of a clinical response to trastuzumab in P04626 -driven tumors . To test this hypothesis , we generated nontransformed human MCF10A mammary epithelial cells stably expressing a chimeric P04626 -FKBP molecule that could be conditionally induced to homodimerize by adding the FKBP ligand AP1510 , or instead induced to heterodimerize with P00533 or P21860 by adding the heterodimer ligands P01133 /TGFα or heregulin . AP1510 , P01133 , and heregulin each induced growth of MCF10A cells expressing P04626 -FKBP . DB00072 inhibited homodimer-mediated but not heterodimer-mediated cell growth . In contrast , the P04626 antibody pertuzumab , which blocks P04626 heterodimerization , inhibited growth induced by heregulin but not AP1510 . Lastly , the P04626 / P00533 tyrosine kinase inhibitor lapatinib blocked both homodimer- and heterodimer-induced growth . AP1510 triggered phosphorylation of Erk1/2 but not AKT , whereas trastuzumab inhibited AP1510-induced Erk1/2 phosphorylation and Shc- P04626 homodimer binding , but not TGFα-induced AKT phosphorylation . Consistent with these observations , high levels of P04626 homodimers correlated with longer time to progression following trastuzumab therapy in a cohort of patients with P04626 -overexpressing breast cancer . Together , our findings confirm the notion that P04626 oligomeric states regulate P04626 signaling , also arguing that trastuzumab sensitivity of homodimers may reflect their inability to activate the PI3K ( phosphoinositide 3-kinase ) /AKT pathway . A clinical implication of our results is that high levels of P04626 homodimers may predict a positive response to trastuzumab . Dynamic simulations of pathways downstream of P00533 -family , including mutations and treatments : concordance with experimental results . The pathways downstream of ErbB-family proteins are very important in BC , especially when considering treatment with onco-protein inhibitors . We studied and implemented dynamic simulations of four downstream pathways and described the fragment of the signaling network we evaluated as a Molecular Interaction Map . Our simulations , enacted using Ordinary Differential Equations , involved 242 modified species and complexes , 279 reversible reactions and 111 catalytic reactions . Mutations within a single pathway tended to be mutually exclusive ; only inhibitors acting at , or downstream ( not upstream ) , of a given mutation were active . A double alteration along two distinct pathways required the inhibition of both pathways . We started an analysis of sensitivity/robustness of our network , and we systematically introduced several individual fluctuations of total concentrations of independent molecular species . Only very few cases showed significant sensitivity . We transduced the ErbB2 over-expressing BC line , BT474 , with the P01112 ( V12 ) mutant , then treated it with ErbB-family and phosphorylated MEK ( MEKPP ) inhibitors , DB01259 and U0126 , respectively . Experimental and simulation results were highly concordant , showing statistical significance for both pathways and for two respective endpoints , i.e. phosphorylated active forms of P29323 and Akt , p one tailed = .0072 and = .0022 , respectively . Working with a complex 39 basic species signaling network region , this technology facilitates both comprehension and effective , efficient and accurate modeling and data interpretation . Dynamic network simulations we performed proved to be both practical and valuable for a posteriori comprehension of biological networks and signaling , thereby greatly facilitating handling , and thus complete exploitation , of biological data . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Characterization of bortezomib-adapted I-45 mesothelioma cells . BACKGROUND : DB00188 , a proteasome-specific inhibitor , has emerged as a promising cancer therapeutic agent . However , development of resistance to bortezomib may pose a challenge to effective anticancer therapy . Therefore , characterization of cellular mechanisms involved in bortezomib resistance and development of effective strategies to overcome this resistance represent important steps in the advancement of bortezomib-mediated cancer therapy . RESULTS : The present study reports the development of I-45-BTZ-R , a bortezomib-resistant cell line , from the bortezomib-sensitive mesothelioma cell line I-45 . I-45-BTZ-R cells showed no cross-resistance to the chemotherapeutic drugs cisplatin , 5-fluorouracil , and doxorubicin . Moreover , the bortezomib-adapted I-45-BTZ-R cells had decreased growth kinemics and did not over express proteasome subunit beta5 ( P28074 ) as compared to parental I-45 cells . I-45-BTZ-R cells and parental I-45 cells showed similar inhibition of proteasome activity , but I-45-BTZ-R cells exhibited much less accumulation of ubiquitinated proteins following exposure to 40 nm bortezomib . Further studies revealed that relatively low doses of bortezomib did not induce an unfolded protein response ( UPR ) in the bortezomib-adapted cells , while higher doses induced UPR with concomitant cell death , as evidenced by higher expression of the mitochondrial chaperone protein Bip and the endoplasmic reticulum ( ER ) stress-related pro-apoptotic protein P35638 . In addition , bortezomib exposure did not induce the accumulation of the pro-apoptotic proteins p53 , Mcl-1S , and noxa in the bortezomib-adapted cells . CONCLUSION : These results suggest that UPR evasion , together with reduced pro-apoptotic gene induction , accounts for bortezomib resistance in the bortezomib-adapted mesothelioma cell line I-45-BTZ-R .

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Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . The 80th anniversary of vitamin E : beyond its antioxidant properties . Molecules provided with an antioxidant function may have additional properties , the latter being sometimes of greater importance than the former . In the last ten years , DB00163 has revealed precise cellular functions , some of which are independent of its antioxidant/radical scavenging ability . At the posttranslational level , DB00163 inhibits protein kinase C and P09917 and activates protein phosphatase 2A and diacylglycerol kinase . Some genes ( P16671 , alpha-TTP , alpha-tropomyosin , and collagenase ) are affected by DB00163 at the transcriptional level . alpha-Tocopherol also induces inhibition of cell proliferation , platelet aggregation and monocyte adhesion . These effects are unrelated to the antioxidant activity of vitamin E , but rather are believed to be a result of specific interactions of vitamin E with components of the cell , e. g. proteins , enzymes and membranes . This review focuses on novel non-antioxidant functions of DB00163 and discusses the possibility that many of the effects previously attributed to the antioxidant functions can also be explained by non-antioxidant mechanisms . Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . Identification of regions of leukotriene C4 synthase which direct the enzyme to its nuclear envelope localization . Leukotrienes ( LTs ) are fatty acid derivatives formed by oxygenation of arachidonic acid via the P09917 ( P09917 ) pathway . Upon activation of inflammatory cells P09917 is translocated to the nuclear envelope ( NE ) where it converts arachidonic acid to the unstable epoxide P09960 . P09960 is further converted to LTC4 by conjugation with glutathione , a reaction catalyzed by the integral membrane protein Q16873 ( Q16873 ) , which is localized on the NE and endoplasmic reticulum ( ER ) . We now report the mapping of regions of Q16873 that are important for its subcellular localization . Multiple constructs encoding fusion proteins of green fluorescent protein ( GFP ) as the N-terminal part and various truncated variants of human Q16873 as C-terminal part were prepared and transfected into P29320 293/T or COS-7 cells . Constructs encoding hydrophobic region 1 of Q16873 ( amino acids 6-27 ) did not give distinct membrane localized fluorescence . In contrast hydrophobic region 2 ( amino acids 60-89 ) gave a localization pattern similar to that of full length Q16873 . Hydrophobic region 3 ( amino acids 114-135 ) directed GFP to a localization indistinguishable from that of full length Q16873 . A minimal directing sequence , amino acids 117-132 , was identified by further truncation . The involvement of the hydrophobic regions in the homo-oligomerization of Q16873 was investigated using bioluminescence resonance energy transfer ( BRET ) analysis in living cells . BRET data showed that hydrophobic regions 1 and 3 each allowed oligomerization to occur . These regions most likely form transmembrane helices , suggesting that homo-oligomerization of Q16873 is due to helix-helix interactions in the membrane . Cytokine-modulating activity of tepoxalin , a new potential antirheumatic . Tepoxalin is a new dual cyclooxygenase/ P09917 anti-inflammatory compound currently under clinical investigation . It has been shown to possess anti-inflammatory activity in a variety of animal models and more recently to inhibit P60568 induced signal transduction . The current study was conducted to evaluate the cytokine modulating activity of tepoxalin and the role of iron in these effects . In human peripheral blood mononuclear cells ( PBMC ) stimulated with OKT3/PMA , tepoxalin inhibited lymphocyte proliferation with an IC50 of 6 microM . Additionally , it inhibited the production of LTB4 ( IC50 = 0.5 microM ) and the cytokines P60568 , P05231 and P01375 alpha ( IC50 = 10-12 microM ) . Cytotoxicity was not demonstrated at these concentrations . Add-back experiments with either cytokines ( P60568 or P05231 ) , LTB4 or conditioned media failed to restore the proliferative response in the presence of tepoxalin . However , the concurrent addition of iron ( in the form of ferrous or ferric chloride and other iron salts ) reversed the inhibition of proliferation caused by tepoxalin . Tepoxalin also inhibits the activation of NF kappa B , a transcription factor which acts on several cytokine genes . Tepoxalin 's effect on NF kappa B is also reversed by the addition of iron salts . These data suggest that the action of tepoxalin to inhibit proliferation in PBMC may be at least in part due to its ability to reduce the amount of available iron resulting in decreased activation of NF kappa B and subsequent inhibition of cytokine production . Cytoprotective properties of DB00163 are related to gene regulation in cultured D-galactosamine-treated human hepatocytes . DB00163 ( DB00163 ) has demonstrated antioxidant activity and gene-regulatory properties . d-Galactosamine ( D-GalN ) -induced cell death is mediated by nitric oxide in hepatocytes , and it is associated with hepatic steatosis . The beneficial properties of DB00163 and their relation to oxidative stress and gene regulation were assessed in D-GalN-induced cell death . Hepatocytes were isolated from human liver resections by a collagenase perfusion technique . alpha-Tocopherol ( 50 microM ) was administered at the advanced stages ( 10 h ) of D-GalN-induced cell death in cultured hepatocytes . Cell death , oxidative stress , DB00163 metabolism , and NF-kappaB- , pregnane X receptor ( O75469 ) - , and peroxisome proliferator-activated receptor ( Q07869 ) -associated gene regulation were estimated in the hepatocytes . D-GalN increased cell death and DB00163 metabolism . alpha-Tocopherol exerted a moderate beneficial effect against apoptosis and necrosis induced by D-GalN . Induction ( rifampicin ) or inhibition ( ketoconazole ) of DB00163 metabolism and overexpression of O75469 showed that the increase in O75469 -related P08684 expression caused by DB00163 enhanced cell death in hepatocytes . Nevertheless , the reduction in NF-kappaB activation and inducible nitric oxide synthase expression and the enhancement of Q07869 and carnitine palmitoyl transferase gene expression by DB00163 may be relevant for cell survival . In conclusion , the cytoprotective properties of DB00163 are mostly related to gene regulation rather than to antioxidant activity in toxin-induced cell death in hepatocytes . Association between Q5S007 and Q13541 protein levels in normal and malignant cells . Translational control is a crucial component of cancer development and progression . Eukaryotic initiation factor ( eIF ) 4E mediates eIF4F association with the mRNA 5' cap structure to stimulate cap-dependent translation initiation . The P06730 -binding protein , Q13541 , regulates cap-dependent translation through its phosphorylation at multiple sites . It has been described that some human carcinomas present a high level of p- Q13541 , not always associated with high levels of p- P42345 . These previous observations suggest that other kinases could be involved in Q13541 phosporylation . Investigation in new kinases that could be implicated in Q13541 phosphorylation and mechanisms that affect Q13541 stability is important to understand the role of P06730 in cell transformation . In this study , we examined 48 kinases that could be involved in Q13541 phosphorylation and stability . The screening study was based on analysis of Q13541 status after inhibition of these kinases in a breast carcinoma cell line . Several kinases affecting Q13541 stability ( Q5S007 , RAF-1 , p38γ , GSK3β , AMPKα , PRKACA and P22694 ) and Q13541 phosphorylation ( P06493 , PDK1 , P12931 , P05771 , Q13177 , p38β , P17252 and CaMKKB ) were identified . These findings provide evidence that Q13541 can be regulated and stabilized by multiple kinases implicated in several cell signaling pathways . We focus on the finding that Q5S007 down-regulation was associated with a clearly decreased Q13541 protein ( and not with mRNA down-regulation ) . Importantly , knockdown of Q5S007 associated with high proliferative rate in normal cells and treatment with rapamycin and/or proteosome inhibition suppressed Q13541 protein degradation . These results offer new insights into the regulation of total and phosphorylated Q13541 . Gamma-tocopherol , but not DB00163 , decreases proinflammatory eicosanoids and inflammation damage in rats . Gamma-tocopherol ( gammaT ) , the major form of vitamin E in U.S. diets , and its physiological metabolite 2 , 7 , 8-trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman ( gamma-CEHC ) , in contrast to DB00163 ( alphaT ) , the primary vitamin E in supplements , inhibit cyclooxygenase-catalyzed synthesis of prostaglandin E2 ( DB00917 ) in activated macrophages and epithelial cells . Here we report that in carrageenan-induced inflammation in male Wistar rats , administration of gammaT ( 33 or 100 mg/kg ) and gamma-CEHC ( 2 mg/pouch ) , but not alphaT ( 33 mg/kg ) , significantly reduced DB00917 synthesis at the site of inflammation . gammaT , but not alphaT , significantly inhibited the formation of leukotriene B4 , a potent chemotactic agent synthesized by the P09917 of neutrophils . Although gammaT had no effect on neutrophil infiltration , it significantly attenuated the partial loss of food consumption caused by inflammation-associated discomfort . Administration of gammaT led consistently to a significant reduction of inflammation-mediated increase in 8-isoprostane , a biomarker of lipid peroxidation . gammaT at 100 mg/kg reduced P01375 ( 65 % ;P=0.069 ) , total nitrate/nitrite ( 40 % ;P=0.1 ) , and lactate dehydrogenase activity ( 30 % ;P=0.067 ) . Collectively , gammaT inhibits proinflammatory DB00917 and LTB4 , decreases P01375 , and attenuates inflammation-mediated damage . These findings provide strong evidence that gammaT shows anti-inflammatory activities in vivo that may be important for human disease prevention and therapy . Cloning of a novel phosphatidylinositol kinase-related kinase : characterization of the human Q96Q15 RNA surveillance protein . We have cloned and characterized a new member of the phosphatidylinositol kinase ( PIK ) -related kinase family . This gene , which we term human Q96Q15 ( Q96Q15 ) , is orthologous to Caenorhabditis elegans Q96Q15 , a protein that functions in nonsense-mediated mRNA decay ( Q53H76 ). cDNA sequencing revealed that Q96Q15 encodes a protein of 3031 amino acids containing a conserved kinase domain , a C-terminal domain unique to the PIK-related kinases and an P62942 -rapamycin binding-like domain similar to that found in the PIK-related kinase P42345 . Immunopurified FLAG-tagged Q96Q15 exhibits protein kinase activity as measured by autophosphorylation and phosphorylation of the generic PIK-related kinase substrate PHAS-1. Q96Q15 kinase activity is inhibited by high nanomolar concentrations of wortmannin ( IC(50) = 105 nm ) but is not inhibited by a P62942 -rapamycin complex . Mutation of conserved residues within the kinase domain of Q96Q15 abolishes both autophosphorylation and substrate phosphorylation , demonstrating that Q96Q15 exhibits intrinsic protein kinase activity . Q96Q15 phosphorylates purified Q92900 protein , a phosphoprotein that plays a critical role in Q53H76 , at sites that are also phosphorylated in whole cells . Based on these data , we conclude that Q96Q15 is the human orthologue to C. elegans Q96Q15 . Our data indicate that Q96Q15 may function in Q53H76 by directly phosphorylating Q92900 protein at physiologically relevant sites . Genome-wide association study identifies common variants associated with circulating vitamin E levels . In genome-wide association studies ( GWAS ) of common genetic variants associated with circulating alpha- and gamma-tocopherol concentrations in two adult cohorts comprising 5006 men of European descent , we observed three loci associated with DB00163 levels , two novel single-nucleotide polymorphisms ( SNPs ) , rs2108622 on 19pter-p13.11 ( P= 1.7 × 10(-8) ) and rs11057830 on 12q24.31 ( P= 2.0 × 10(-8) ) and confirmed a previously reported locus marked by rs964184 on 11q23.3 ( P= 2.7 × 10(-10) ) . The three SNPs have been reported to be associated with lipid metabolism and/or regulation . We replicated these findings in a combined meta-analysis with two independent samples , P= 7.8 × 10(-12) ( rs964184 on 11q23.3 near Q9BRD0 , O75312 and P02647 / P01024 /A4/A5 ) , P= 1.4 × 10(-10) ( rs2108622 on 19pter-p13.11 near P78329 ) and P= 8.2 × 10(-9) ( rs11057830 on 12q24.31 near Q8WTV0 ) . Combined , these SNPs explain 1.7 % of the residual variance in log DB00163 levels . In one of the two male GWAS cohorts ( n= 992 ) , no SNPs were significantly associated with gamma-tocopherol concentrations after including data from the replication sample for 71 independent SNPs with P < 1 × 10(-4) identified . Selective inhibition of histone deacetylase 6 ( Q9UBN7 ) induces DNA damage and sensitizes transformed cells to anticancer agents . Q9UBN7 ( Q9UBN7 ) is structurally and functionally unique among the 11 human zinc-dependent histone deacetylases . Here we show that chemical inhibition with the Q9UBN7 -selective inhibitor tubacin significantly enhances cell death induced by the topoisomerase II inhibitors etoposide and doxorubicin and the pan-HDAC inhibitor DB02546 ( vorinostat ) in transformed cells ( LNCaP , MCF-7 ) , an effect not observed in normal cells ( human foreskin fibroblast cells ) . The inactive analogue of tubacin , nil-tubacin , does not sensitize transformed cells to these anticancer agents . Further , we show that down-regulation of Q9UBN7 expression by shRNA in LNCaP cells enhances cell death induced by etoposide , doxorubicin , and DB02546 . Tubacin in combination with DB02546 or etoposide is more potent than either drug alone in activating the intrinsic apoptotic pathway in transformed cells , as evidenced by an increase in PARP cleavage and partial inhibition of this effect by the pan-caspase inhibitor Z-VAD-fmk . Q9UBN7 inhibition with tubacin induces the accumulation of γ P16104 , an early marker of DNA double-strand breaks . Tubacin enhances DNA damage induced by etoposide or DB02546 as indicated by increased accumulation of γ P16104 and activation of the checkpoint kinase Chk2 . Tubacin induces the expression of P35638 ( P35638 / P35638 ) , a transcription factor up-regulated in response to cellular stress . P35638 induction is further increased when tubacin is combined with DB02546 . These findings point to mechanisms by which Q9UBN7 -selective inhibition can enhance the efficacy of certain anti-cancer agents in transformed cells . The effect of DB00163 on monocyte proatherogenic activity . Atherosclerosis is the leading cause of morbidity and mortality in Westernized populations . The monocyte is a crucial cell in the genesis of the atherosclerotic lesion and is present during all stages of atherosclerosis . alpha-Tocopherol ( AT ) is the most active component of the vitamin E family and is the principal and most potent lipid-soluble antioxidant in plasma and LDL . With regard to monocyte function , AT supplementation ( 1200 IU/d ) has been shown to decrease release of reactive oxygen species , lipid oxidation , release of cytokines such as interleukin-1ss ( IL-1ss ) and tumor necrosis factor-alpha ( P01375 ) and decrease adhesion of monocytes to human endothelium . The mechanism of inhibition of superoxide and lipid oxidation by monocytes appears to be via inhibition of protein kinase C ( PKC ) , the decrease in IL-1ss and P01375 release by inhibition of P09917 and the inhibition of monocyte-endothelial cell adhesion via decrease in adhesion molecules on monocytes , CD11b and VLA-4 and by decreasing DNA-binding activity of nuclear transcription factor kappaB . Thus , in addition to the decrease in oxidative stress resulting from AT supplementation , as evidenced by decreased F(2)-isoprostanes and LDL oxidizability , AT is anti-inflammatory and exerts beneficial antiatherogenic effects on cells crucial in atherogenesis such as monocytes . DB00163 , P09917 and oxidative stress in haemodialysis patients : facts , not fancies . Red blood cell surface adhesion molecules : their possible roles in normal human physiology and disease . Human erythrocytes express a relatively large number of known adhesion receptors , despite the fact that red blood cells ( RBCs ) are generally considered to be nonadhesive for endothelial cell surfaces . Some of these adhesion receptors are expressed by many other tissues , while others have more limited tissue distribution . Some adhesion receptors , including P16671 and VLA-4 , are only expressed by immature erythroid cells , while others are present on mature erythrocytes . The structure and function of these proteins is reviewed here . LW , P16671 , P19256 , and CD147 have been shown in other tissues to mediate cell-cell interaction . Other receptors , such as P16070 , VLA-4 , and B- P62158 /LU , can mediate adhesion to components of extracellular matrix . In addition , their roles in normal erythropolesis , as well as in the pathophysiology of human disease , are summarized . The most convincing evidence for a pathophysiologic role for any of these receptors on erythrocytes comes from studies of cells from patients homozygous for hemoglobin S , as RBC adhesion is thought to contribute to vaso-occlusion . Thus , receptors such as B- P62158 /LU may become targets for future therapy aimed at preventing or ameliorating this thrombotic process . G protein-coupled receptor-induced sensitization of phospholipase C stimulation by receptor tyrosine kinases . Activation of stably expressed M(2) and M(3) muscarinic acetylcholine receptors ( mAChRs ) as well as of endogenously expressed lysophosphatidic acid and purinergic receptors in P29320 -293 cells can induce a long lasting potentiation of phospholipase C ( P98160 ) stimulation by these and other G protein-coupled receptors ( GPCRs ) . Here , we report that GPCRs can induce an up-regulation of P98160 stimulation by receptor tyrosine kinases ( RTKs ) as well and provide essential mechanistic characteristics of this sensitization process . Pretreatment of P29320 -293 cells for 2 min with carbachol , a mAChR agonist , lysophosphatidic acid , or DB00171 , followed by agonist washout , strongly increased ( by 2-3-fold ) maximal P98160 stimulation ( measured >/= 40 min later ) by epidermal growth factor and platelet-derived growth factor , but not insulin , and largely enhanced P98160 sensitivity to these RTK agonists . The up-regulation of RTK-induced P98160 stimulation was cycloheximide-insensitive and was observed for up to approximately 90 min after removal of the GPCR agonist . Sensitization of receptor-induced P98160 stimulation caused by prior M(2) mAChR activation was fully prevented by pertussis toxin and strongly reduced by expression of Gbetagamma scavengers . Furthermore , inhibition of conventional protein kinase C ( PKC ) isoenzymes and chelation of intracellular Ca(2+) suppressed the sensitization process , while overexpression of P17252 , but not PKC-betaI , further enhanced the M(2) mAChR-induced sensitization of P98160 stimulation . None of these treatments affected acute P98160 stimulation by either GPCR or RTK agonists . Taken together , short term activation of GPCRs can induce a strong and long lasting sensitization of P98160 stimulation by RTKs , a process apparently involving G(i)-derived Gbetagammas as well as increases in intracellular Ca(2+) and activation of a PKC isoenzyme , most likely P17252 . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Novel cinnamyl hydroxyamides and 2-aminoanilides as histone deacetylase inhibitors : apoptotic induction and cytodifferentiation activity . Four novel series of cinnamyl-containing histone deacetylase ( HDAC ) inhibitors 1-4 are described , containing hydroxamate ( 1 and 3 ) or 2-aminoanilide ( 2 and 4 ) derivatives . When screened against class I ( maize HD1-B and human Q13547 ) and class II ( maize HD1-A and human P56524 ) HDACs , most hydroxamates and 2-aminoanilides displayed potent and selective inhibition toward class I enzymes . Immunoblotting analyses performed in U937 leukemia cells generally revealed high acetyl-H3 and low acetyl-α-tubulin levels . Exceptions are compounds 3 f-i , 3 m-o , and 4 k , which showed higher tubulin acetylation than DB02546 . In U937 cells , cell-cycle blockade in either the G₂/M or G₁/S phase was observed with 1-4 . Five hydroxamates ( compounds 1 h-l ) effected a two- to greater than threefold greater percent apoptosis than DB02546 , and in the CD11c cytodifferentiation test some 2-aminoanilides belonging to both series 2 and 4 were more active than MS-275 . The highest-scoring derivatives in terms of apoptosis ( 1 k , 1 l ) or cytodifferentiation ( 2 c , 4 n ) also showed antiproliferative activity in U937 cells , thus representing valuable tools for study in other cancer contexts . Oxidized LDL and lysophosphatidylcholine stimulate plasminogen activator inhibitor-1 expression through reactive oxygen species generation and P27361 /2 activation in 3T3- Q9NUQ9 adipocytes . P00747 activator inhibitor-1 ( P05121 ) is secreted from adipose tissue and is considered to be a risk factor for both atherosclerosis and insulin resistance . Here we report for the first time that P05121 expression is enhanced by oxidized low-density lipoprotein ( OxLDL ) and its lipid component lysophosphatidylcholine ( Q16549 ) in mouse 3T3- Q9NUQ9 adipocytes . In fully differentiated 3T3- Q9NUQ9 cells , OxLDL treatment increased the mRNA expression and protein secretion of P05121 in a dose- and time-dependent manner , whereas native LDL had no effect . The addition of an anti- P16671 antibody suppressed OxLDL-stimulated P05121 expression by 50 % , suggesting that adipose-derived P16671 contributes to roughly half of the P05121 expression stimulated by OxLDL . In addition , pharmacological experiments showed that the OxLDL-stimulated enhancement in P05121 expression was mediated through the generation of reactive oxygen species ( ROS ) and phosphorylation of extracellular signal-regulated kinase 1/2 . Furthermore , Q16549 , a major lipid component of OxLDL , was responsible for the enhanced expression of P05121 as phospholipase A(2)-treated acetyl LDL , which generates Q16549 , strongly stimulated P05121 expression , whereas acetyl LDL itself had no such activity . These data demonstrate that the uptake of OxLDL and , in particular , its lipid component Q16549 into adipocytes triggers aberrant ROS-mediated P05121 expression , which may be involved in the pathogenesis of metabolic syndrome . Mechanisms of epidermal growth factor-induced contraction of guinea pig airways . We investigated the functional effects of epidermal growth factor ( P01133 ) on guinea pig airways in vitro . P01133 ( 3 ng/ml to 1 microgram/ml ) induced a concentration-dependent contraction in epithelium-denuded strips . The average maximal contraction was 0.64 +/- 0.1 g ( mean +/- S.E. , for n = 27 ) , which was 72.0 +/- 9.5 % of the 100 mM DB00761 -induced contraction . The EC50 was 12.3 +/- 1.6 ng/ml . The presence of the epithelium significantly suppressed the P01133 -induced contraction ( P < 0.01 ) . P01133 -induced contraction was abolished by cyclooxygenase inhibitors ( indomethacin and ibuprofen ) and a P09917 inhibitor , 2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-1,4-benz oqu inone ( AA-861 ) . It was also inhibited by a leukotriene-receptor antagonist , 8-[p-(4-phenylbutyloxy)benzoyl]amino-2-(tetrazol-5-yl)-4-oxo -4H-1-benzopyran hemihydrate ( ONO-1078 ) but not affected by a thromboxane A2-synthetase inhibitor , ( E ) -3-[4-(1-imidazolylmethyl)phenyl]-2-propenoic acid ( OKY-046 ) or a thromboxane A2-receptor antagonist , 9,11-epithio-11,12-methano-thromboxane A2 ( ONO-3708 ) . A phospholipase A2 inhibitor ( mepacrine ) inhibited the P01133 -induced contraction but a diacylglycerol-lipase inhibitor , 1,6-di- ( O-(carbamoyl)cyclohexanone oxime ) hexane ( U-57908 ) and a phospholipase D inhibitor ( wortmannin ) did not affect it . A tyrosine kinase inhibitor ( genistein ) abolished it . Measurement of prostanoids showed that P01133 ( 300 ng/ml ) did not increase the prostaglandin F2 alpha level in either epithelium-intact or epithelium-denuded strips . In epithelium-intact strips , P01133 significantly increased the prostaglandin E2 concentration ( P < 0.01 ) . These results suggest that P01133 causes contraction of guinea pig airway smooth muscle by activating tyrosine kinase followed by phospholipase A2 activation , and that arachidonic acid metabolites , especially leukotrienes , may have important roles in this contraction . Arachidonate cascade , apoptosis , and vitamin E in peripheral blood mononuclear cells from hemodialysis patients . BACKGROUND : Lipid peroxidation and oxidative stress are enhanced in peripheral blood mononuclear cells ( PBMCs ) from hemodialysis ( HD ) patients because of upregulation of the P09917 pathway of the arachidonate cascade . 5-Lipoxygenase activity is specifically inhibited by vitamin E both in vitro and in vivo regardless of its administration route . METHODS : The effect of arachidonate cascade enzymes and vitamin E on oxidative stress and apoptosis was investigated in PBMCs from 16 maintenance HD patients treated for at least 6 months with cuprammonium rayon membranes in a two-step crossover study : after a 4-week treatment with vitamin E-coated cuprammonium rayon membranes and again after a 4-week treatment with oral vitamin E. Control PBMCs were obtained from 16 healthy volunteers . RESULTS : Membrane lipoperoxidation , cellular luminescence , membrane fluidity , and leukotriene B(4) content were significantly greater in PBMCs from HD patients ; lipoxygenase was upregulated , but prostaglandin H synthase ( P61457 ) was not affected . Regardless of administration route , vitamin E partially controlled lipid peroxidation and oxidative stress through direct inhibition of P09917 . Cultured PBMCs from HD patients showed a significant increase in apoptotic cells compared with controls . DB00163 markedly reduced cell luminescence , membrane fluidity , and apoptosis , whereas the P61457 inhibitor indomethacin was ineffective . Similar results were obtained with control PBMCs induced to apoptosis by hydrogen peroxide . CONCLUSION : Reported data suggest that the P09917 branch of the arachidonate cascade is only responsible for membrane peroxidation , oxidative stress , and apoptosis of PBMCs of HD patients , and administration of vitamin E may be helpful in the control of oxidative stress-related disease in these subjects . Effect of the hemoregulatory peptide (pEEDCK)2 (pyroGlu- DB00142 - DB00128 - DB00151 -Lys)2 and MIP-1alpha is reduced in bone marrow cultures from patients with chronic myeloid leukemia ( CML ) . The granulocyte-derived hemoregulatory peptide pyroGlu- DB00142 - DB00128 - DB00151 -Lys = pEEDCK is known to keep hematopoietic cells quiescent . When oxidized to its dimeric form (pEEDCK)2 , it activates growth of hematopoietic progenitors in association with stroma-derived cytokines . (pEEDCK)2 has a DB00151 - DB00151 motif which is also a typical feature of the macrophage inflammatory protein ( MIP-1alpha ) . The present study was designed to analyze differences between the response of normal and leukemic progenitor cells to (pEEDCK)2 or MIP-1alpha . When long-term bone marrow cultures ( LTBMCs ) were incubated with (pEEDCK)2 or MIP-1alpha and/or cytokines , the stimulatory effect on colony-forming units-granulocyte/erythroid/macrophage/megakaryocyte of LTBMC from chronic myeloid leukemia ( CML ) patients was less than 50 % compared to LTBMC from healthy humans . No difference in oncogene expression could be observed in LTBMC from CML patients regarding reduction of Philadelphia chromosome-associated transcription of the P11274 - P00519 gene . With respect to the expression of growth and differentiation-associated genes ( Galpha16 , P09917 , phospholipaseA2 , c-kit , and P28906 ) , which were analyzed from LTBMC by semiquantitative reverse transcriptase-polymerase chain reaction , the same transcription rate was observed in CML patients and healthy donors . However , two isoforms of a key enzyme of oxidative metabolism , carnitine palmitoyltransferase ( P50416 and Q92523 ) , showed 50-fold higher expression rates in LTBMC cells of healthy donors compared to CML patients . It is known that a decrease in oxidative metabolism is associated with an increase in redox equivalents in malignancy . This might result in a reduction of disulphide bonds in (pEEDCK)2 or MIP-1alpha , thus inducing a downregulation of these factors in bone marrow from CML patients . Specific cellular responses to DB00163 . In the last 10 years precise cellular functions of DB00163 , some of which are independent of its antioxidant/radical-scavenging ability , have been revealed . Absorption of DB00163 from the gut is a selective process . Other tocopherols are not absorbed or are absorbed to a lesser extent . At the post-translational level , DB00163 inhibits protein kinase C and P09917 and activates protein phosphatase 2A and diacylglycerol kinase . Some genes [ platelet glycoprotein IV/thrombospondin receptor/class B scavenger receptor ( P16671 ) , DB00163 transfer protein ( alpha-TTP ) , alpha-tropomyosin , connective tissue growth factor and collagenase ] are affected by DB00163 at the transcriptional level . alpha-Tocopherol also inhibits cell proliferation , platelet aggregation , monocyte adhesion and the oxygen burst in neutrophils . Other antioxidants , such as beta-tocopherol and probucol , do not mimic these effects , suggesting a nonantioxidant , DB00163 -specific molecular mechanism . Alpha-tocopherol as a modulator of smooth muscle cell proliferation . The effects of DB00163 and beta-tocopherol have been studied in rat and human aortic smooth muscle cells . Alpha-tocopherol , but not beta-tocopherol , inhibited smooth muscle cell proliferation and protein kinase C in a dose-dependent manner , at concentrations ranging from 10 to 50 microM . Beta-tocopherol added simultaneously with DB00163 prevented both proliferation and protein kinase C inhibition . Protein kinase C inhibition was cell cycle-dependent and it was prevented by okadaic acid , a protein phosphatase inhibitor . Protein kinase C activity measured from aortas of cholesterol-fed rabbits was also inhibited by DB00163 . By using protein kinase C ( PKC ) isoform-specific inhibitors and immunoprecipitation reactions it was found that P17252 was selectively inhibited by DB00163 . Further , an activation of protein phosphatase 2A by DB00163 was found , which caused P17252 dephosphorylation and inhibition . Ultimately , this cascade of events at the level of cell signal transduction leads to the inhibition of smooth muscle cell proliferation . Activation of P09917 and related cell membrane lipoperoxidation in hemodialysis patients . Lipid peroxidation was shown at the membrane level in peripheral blood cells of patients hemodialyzed on cuprophan dialyzers , and was mainly attributable to the generation of conjugated hydroperoxides in the lipid bilayer . The oxidative index ( i.e. , the A234/205 ratio ) of membrane lipids was 3.2-fold higher in hemodialysis patients than in healthy control subjects , and also the level of leukotriene B4 was significantly increased ( up to 1.7-fold over control ) . Both membrane peroxidation and release of leukotriene B4 were linked to upregulation of P09917 activity ( up to 2.4-fold over control ) and expression at the protein level ( up to 1.9-fold ) . DB00163 , the most important lipophilic antioxidant , prevented both membrane peroxidation and release of leukotriene B4 by inhibiting P09917 activity without affecting enzyme expression . Similar results were observed in patients hemodialyzed on polymethylmetacrylate membranes , but in this case the activation of P09917 was less pronounced . The use of a purified P09917 demonstrated that vitamin E was a reversible inhibitor of enzyme activity ( IC50 = 35 +/- 4 microM ) , further characterized as noncompetitive ( Ki = 30 +/- 3 microM ) . Taken together , the results reported here shed some light on the mechanism responsible for the oxidative damage in hemodialysis . Moreover , the beneficial effect of vitamin E described here may have relevance for the therapy of patients with kidney disease . Elevated retinol binding protein 4 induces apolipoprotein B production and associates with hypertriglyceridemia . CONTEXT AND OBJECTIVE : A high level of retinol binding protein 4 ( P02753 ) is reported to be associated with insulin resistance in humans . However , evidence from large-scale populations about the relationship between serum P02753 and metabolic phenotypes is scarce . In the present study , we aimed to evaluate serum P02753 distribution and its association with metabolic phenotypes among middle-aged and elderly Chinese . DESIGN AND PARTICIPANTS : Serum concentrations of P02753 in a cross-sectional sample of 2780 Chinese population aged 50-70 years old in Guangzhou were measured by ELISA . RESULTS : The mean of serum P02753 concentration was 28.04 μg/mL for male and 37.76 μg/mL for female ( P < .01 ) , respectively . Circulating P02753 was positively correlated with serum triglyceride and apolipoprotein B ( apoB ) concentrations . The odds ratio ( OR ) was substantially higher for hypertriglyceridemia ( OR , 3.26 ; 95 % confidence interval , 2.36-4.51 ) in the highest P02753 quartile compared with those in the lowest quartile after multiple adjustment for confounders . Furthermore , serum P02753 was significantly associated with fasting glucose , insulin levels , and homeostasis model assessment index-insulin resistance ( HOMA-IR ) . Moreover , we showed that P02753 enhanced microsomal triglyceride transfer protein ( P55157 ) expression and activity via up-regulation of protein disulfide isomerase ( P07237 ) , suppressed low-density lipoprotein receptor ( P01130 ) expression , and impaired insulin-signaling pathway , leading to inductions in apoB secretion both in vitro and in vivo . CONCLUSIONS : Elevated circulating P02753 concentrations were associated with higher risk of hypertriglyceridemia by inducing the secretion of triglyceride-rich apoB-containing lipoproteins . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . DB00163 -related inhibition of monocyte P09917 and cardiovascular outcome in maintenance hemodialysis patients . A daily supplement of vitamin E is recommended for the secondary prevention of cardiovascular events in end-stage renal disease patients on maintenance hemodialysis . DB00163 has been entrusted with therapeutic properties against cardiovascular disease for more than 60 years . Several epidemiological studies and intervention trials have been performed with vitamin E , and some of them showed that it prevents atherosclerosis . For a long time , vitamin E was assumed to act by decreasing the oxidation of low-density lipoproteins , a key step in atherosclerosis initiation . However , at the cellular level vitamin E interferes with smooth muscle cell proliferation , platelet aggregation , monocyte adhesion , and oxidized low-density lipoproteins uptake and cytokine production , all reactions implied in the progression of atherosclerosis . Recent research points out that these effects may be not only the result of the antioxidant activity of vitamin E but also of its distinct molecular actions . These biological properties of vitamin E may allow to design better strategies for primary and secondary prevention of cardiovascular disease , with a potential exploitation of vitamin E supplements in primary and secondary prevention of major adverse cardiovascular events in all uremic patients . In this review , we also outline relevant patents on vitamin E and lipoxygenase inhibitors . Tocotrienols activate the steroid and xenobiotic receptor , O75469 , and selectively regulate expression of its target genes . DB00163 is an essential nutrient with antioxidant activity . DB00163 is comprised of eight members , alpha- , beta- , gamma- , and delta-tocopherols and alpha- , beta- , gamma- , and delta-tocotrienols . All forms of vitamin E are initially metabolized by omega-oxidation , which is catalyzed by cytochrome P450 enzymes . The steroid and xenobiotic receptor ( O75469 ) is a nuclear receptor that regulates drug clearance in the liver and intestine via induction of genes involved in drug and xenobiotic metabolism . We show here that all four tocotrienols specifically bind to and activate O75469 , whereas tocopherols neither bind nor activate . Surprisingly , tocotrienols show tissue-specific induction of O75469 target genes , particularly P08684 . Tocotrienols up-regulate expression of P08684 but not P22309 ( P22309 ) or multidrug resistance protein-1 ( P08183 ) in primary hepatocytes . In contrast , tocotrienols induce P08183 and P22309 but not P08684 expression in intestinal LS180 cells . We found that nuclear receptor corepressor ( NCoR ) is expressed at relatively high levels in intestinal LS180 cells compared with primary hepatocytes . The unliganded O75469 interacts with NCoR , and this interaction is only partially disrupted by tocotrienols . Expression of a dominant-negative NCoR enhanced the ability of tocotrienols to induce P08684 in LS180 cells , suggesting that NCoR plays an important role in tissue-specific gene regulation by O75469 . Our findings provide a molecular mechanism explaining how vitamin supplements affect the absorption and effectiveness of drugs . Knowledge of drug-nutrient interactions may help reduce the incidence of decreased drug efficacy . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . DB00163 suppresses P09917 -mediated oxidative stress in peripheral blood mononuclear cells of hemodialysis patients regardless of administration route . A number of pathological conditions caused by oxidative stress have been reported in uremic patients undergoing maintenance hemodialysis ( HD ) . Enhanced lipid peroxidation was previously observed in peripheral blood mononuclear cells ( PBMCs ) of HD patients . Upregulation of P09917 ( 5-Lox ) activity and protein content with enhanced production of leukotriene B(4) ( Q06643 (4) ) and membrane lipoperoxides was also shown in PBMCs of HD patients . Administration of free vitamin E specifically inhibited 5-Lox activity without affecting gene expression at the protein level . To assess whether oral or intramuscular ( IM ) administration of vitamin E may suppress 5-Lox in HD patients , PBMCs from 16 subjects on maintenance HD therapy for at least 6 months were investigated before and after a short course of IM or oral administration of vitamin E ( 8 patients per group ) . PBMCs from 13 healthy controls were also evaluated and assumed as the reference standard . DB00163 significantly reduced lipid peroxidation , Q06643 (4) content , and 5-Lox activity in PBMCs , whereas 5-Lox gene expression at the protein level was not affected . There were no significant differences in these parameters between patients treated with IM or oral vitamin E. PBMCs of HD patients showed enhanced membrane lipid peroxidation and release of Q06643 (4) , both linked to upregulation of 5- P28300 : 5-Lox activity and related oxidative stress were significantly ( although not completely ) suppressed by vitamin E regardless of the administration route . A synthetic dl-nordihydroguaiaretic acid ( Nordy ) , inhibits angiogenesis , invasion and proliferation of glioma stem cells within a zebrafish xenotransplantation model . The zebrafish ( Danio rerio ) and their transparent embryos represent a promising model system in cancer research . Compared with other vertebrate model systems , we had previously shown that the zebrafish model provides many advantages over mouse or chicken models to study tumor invasion , angiogenesis , and tumorigenesis . In this study , we systematically investigated the biological features of glioma stem cells ( GSCs ) in a zebrafish model , such as tumor angiogenesis , invasion , and proliferation . We demonstrated that several verified anti-angiogenic agents inhibited angiogenesis that was induced by xenografted-GSCs . We next evaluated the effects of a synthetic dl-nordihydroguaiaretic acid compound ( dl-NDGA or " Nordy " ) , which revealed anti-tumor activity against human GSCs in vitro by establishing parameters through studying its ability to suppress angiogenesis , tumor invasion , and proliferation . Furthermore , our results indicated that Nordy might inhibit GSCs invasion and proliferation through regulation of the arachidonate P09917 ( Alox-5 ) pathway . Moreover , the combination of Nordy and a P15692 inhibitor exhibited an enhanced ability to suppress angiogenesis that was induced by GSCs . By contrast , even following treatment with 50 µM Nordy , there was no discernible effect on zebrafish embryonic development . Together , these results suggested efficacy and safety of using Nordy in vivo , and further demonstrated that this model should be suitable for studying GSCs and anti- P56915 drug evaluation . Effects of ellagic Acid on angiogenic factors in prostate cancer cells . BACKGROUND : Several natural antioxidants , including ellagic acid ( EA ) , have been reported to have chemotherapeutic activity in vivo and in vitro settings . Cytochrome P450 ( CYP ) activity and synthesis of both epoxyeicosatrienoic acids ( EETs ) and 20-hydroxy-5,8,11,14-eicosatetraenoic acid ( 20-HETE ) , together with vascular endothelial growth factor ( P15692 ) and heme oxygenase system ( HO ) have emerged as important modulators of tumor growth and metastasis . METHODS : The anti-angiogenic effects of EA were investigated in the human prostatic cancer cell line LnCap . P09601 , P30519 , P51589 and soluble epoxyde hydrolase ( sEH ) expressions were evaluated by western blotting . Levels of P15692 and osteoprotegerin ( O00300 ) were determined in the culture supernatant using an ELISA assay , while CYP mRNAs were determined by qRT-PCR . RESULTS : EA treatment induced a significant decrease ( p < 0.05 ) in P09601 , P30519 and P51589 expression , and in P15692 and O00300 levels . Similarly P51589 , P78329 and CYPA22 mRNAs were significantly ( p < 0.05 ) down-regulated by EA treatment . The decrease in P51589 mRNA was associated with an increase in sEH expression . CONCLUSIONS : RESULTS reported in the present study highlighted the ability of EA to modulate a new pathway , in addition to anti-proliferative and pro-differentiation properties , via a mechanism that involves a decrease in eicosanoid synthesis and a down-regulation of the HO system in prostate cancer . Cerebral protein kinase C and its mRNA level in apolipoprotein E-deficient mice . It is known that protein kinase C ( PKC ) activity may be one of the fundamental cellular changes associated with memory function . P02649 ( apoE ) deficiency causes cholinergic deficits and memory impairment . ApoE-deficient mouse has been employed as a serviceable model for studying the relation between apoE and the memory deficit induced by cholinergic impairment . Brain-fatty acid binding protein ( b-FABP ) might be functional during development of the nervous system . Peroxisome proliferator-activated receptor ( Q07869 ) is involved in the early change in lipid metabolism . We investigated the alterations not only in cerebral PKC activity , but also in the gene expressions of P05771 , brain-FABP and Q07869 in apoE-deficient mice . The results showed that there was a lower cerebral membrane-bound PKC activity in the apoE-deficient mice than in its wild type strain ( C57BL/6 ) . But there were no significant differences in cytosolic PKC activity . P05771 , b-FABP and Q07869 mRNA expressions in cerebrum were lowered in apoE-deficient mice . These findings may be involved in the dysfunction of the brain neurotransmission system in apoE-deficient mouse . Alternatively , these results also suggest that cerebral apoE plays an important role in brain PKC activation by maintaining an appropriate expression of b-FABP and Q07869 mRNAs . Flavonoids inhibit the oxidative modification of low density lipoproteins by macrophages . Low density lipoproteins ( LDL ) can be oxidatively modified in vitro by macrophages and certain other cell types so that macrophages will take them up much faster . This process may be important in the formation of cholesterol-laden foam cells derived from macrophages in atherosclerotic lesions . In this study , we have shown that certain flavonoids , plant constituents found in the diet , are potent inhibitors of the modification of 125I-labelled LDL by macrophages , with IC50 values in the micromolar range ( e.g. morin and fisetin 1 microM ; quercetin and gossypetin 2 microM ) . The potencies of individual flavonoids in inhibiting LDL modification did not correlate with their previously determined potencies as inhibitors of P09917 and cyclo-oxygenase . The modification of LDL by macrophages exhibits a lag period of about 4-6 hr before enhanced uptake is detected . During this time , there is a rapid depletion in its content of DB00163 ( an endogenous antioxidant found in lipoproteins ) followed by a large increase in the level of hydroperoxides . The flavonoids conserved the DB00163 content of LDL and delayed the onset of detectable lipid peroxidation . Flavonoids also inhibited the cell-free oxidation of LDL mediated by CuSO4 . These findings raise the possibility that flavonoids may protect LDL against oxidation in atherosclerotic lesions and may therefore be natural anti-atherosclerotic components of the diet , although this will depend to a large extent on their pharmacokinetics . In vivo expression of classic PKC isoforms in the rat dental follicle as related to tooth eruption . Activation of protein kinase C ( PKC ) can upregulate tooth eruption molecules such as osteoprotegerin ( O00300 ) and vascular endothelial growth factor ( P15692 ) . In this study , we examined the in vivo gene expression of classic isoforms of PKC in the dental follicle of postnatal rats . The expression level of P17252 was significantly reduced at day 3 followed by a gradual return to day 1 level , a profile similar to O00300 expression . The expression of P05771 was the lowest at day 1 followed by elevated levels from day 3 to day 11 . Expression of P05771 is positively correlated with the expression of overall P15692 and VEGF120 . The expression level of P05129 was relatively steady in the postnatal days . Injection of a PKC activator , phorbol 12-myristate 13-acetate ( PMA ) , at late postnatal days , slightly accelerated first mandibular molar eruption . This study suggests that PKC isoforms may be involved in the regulation of tooth eruption . The role of protein kinase C activation in the pathogenesis of diabetic vascular complications . Many vascular diseases in diabetes are known to be associated with the activation of the diacylglycerol ( DAG ) -protein kinase C ( PKC ) pathway . The major source of DAG that is elevated in diabetes is de novo synthesis from glycolytic intermediates . Among the various PKC isoforms , the beta-isoform has been shown to be persistently activated in diabetic animals . Multiple lines of evidence have shown that many vascular alterations in diabetes -- such as a decrease in the activity of Na+-K+-adenosine triphosphatase ( Na+-K+-ATPase ) , and increases in extracellular matrix , cytokines , permeability , contractility , and cell proliferation -- are caused by activation of PKC . Inhibition of PKC by two different kinds of PKC inhibitors , LY333531 , a selective P05771 -isoform inhibitor , and d- DB00163 , were able to prevent or reverse the various vascular dysfunctions in diabetic rats . These results have also provided in vivo evidence that DAG-PKC activation could be responsible for the hyperglycemia-induced vascular dysfunctions in diabetes . Clinical studies are now being performed to clarify the pathogenic roles of the DAG-PKC pathway in developing vascular complications in diabetic patients . Inhibition of P09917 by vitamin E . Purified P09917 from potato tubers was inhibited strongly by vitamin E and its analogs . The inhibition by d- DB00163 was found to be irreversible and non-competitive with respect to arachidonic acid . An IC50 of 5 microM was calculated for d- DB00163 . The inhibition appears to be unrelated to its antioxidant function . Binding studies with 14C-labelled d- DB00163 revealed that there is a strong interaction between vitamin E and P09917 . Tryptic digestion and peptide mapping of P09917 -vitamin E complex indicate that vitamin E binds strongly to a single peptide . These studies suggest that cellular vitamin E levels may have profound influence on the formation of leukotrienes . Eicosanoid signalling pathways in the heart . Myocardial phospholipids serve as primary reservoirs of arachidonic acid ( AA ) , which is liberated through the rate-determining hydrolytic action of cardiac phospholipases A2 ( PLA2s ) . A predominant P04054 in myocardium is calcium-independent phospholipase A2beta ( iPLA2beta ) , which , through its calmodulin ( P62158 ) and DB00171 -binding domains , is regulated by alterations in local cellular Ca2+ concentrations and cardiac bioenergetic status , respectively . Importantly , iPLA2beta has been demonstrated to be activated by ischaemia through elevation of the concentration of myocardial fatty acyl- DB01992 , which abrogates Ca2+/ P62158 -mediated inhibition of iPLA2beta . AA released by P04054 -catalysed hydrolysis of phospholipids serves as a precursor for eicosanoids generated by pathways dependent on cyclooxygenases ( P36551 ) , lipoxygenases ( P28300 ) , and cytochromes P450 ( CYP ) . Eicosanoids initiate and propagate diverse signalling cascades , primarily through their interaction with cellular receptors and ion channels . However , during pathologic states such as ischaemia or congestive heart failure , eicosanoids contribute to multiple maladaptive changes including inflammation , alterations of cellular growth programmes , and activation of multiple transcriptional events leading to the deleterious sequelae of these pathologic states . This review summarizes the central roles of myocardial PLA(2)s in eicosanoid signalling in the heart , the major P36551 , P28300 , and CYP pathways of eicosanoid generation in the myocardium , and the effects of important eicosanoids on receptor- , ion channel- , and transcription-mediated processes that facilitate cardiac hypertrophy , mediate ischaemic preconditioning , and precipitate arrhythmogenesis in response to pathologic stimuli . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Design , synthesis , and biological evaluation of conformationally constrained aci-reductone mimics of arachidonic acid . An efficient and convergent synthesis has been developed for the production of 3,4-dihydroxy-5- [ 4- ( 2- ( ( 2Z ) -hexenyl ) phenyl ) -3-(1Z)-but enyl ] -2 ( 5H ) -furanone ( 12d ) . This hydrophobic antioxidant is a stable conformationally constrained mimic of arachidonic acid ( AA ) ( 1 ) and its respective aci-reductone analogue ( 2 ) . Pd(0)-catalyzed cross-coupling of 5-(3-butynyl)-3,4-dihydroxy-2(5H)-furanone ( 7 ) with 2- ( ( 2Z ) -hexenyl ) iodobenzene ( 8d ) followed by Lindlar catalyzed hydrogenation produces 12d . Butynyl intermediate 7 is prepared from 2-(benzyloxy)-5-deoxyascorbic acid ( 15 ) by iodination ( I2 , PPh3 , Imd ) , iodo substitution with lithium acetylide ethylenediamine complex ( LiAEDA , HMPA , -5 degrees C ) , and benzyl group cleavage ( Ac2O , Pyr , BCl3 ) . The utility of this synthetic method was demonstrated by the synthesis of analogues 10e-k . Biological testing revealed that certain of these antioxidants inhibit both cyclooxygenase ( P36551 ) and P09917 ( P09917 ) with comparable efficacy as reported for aspirin and zileuton , respectively . The antioxidant activity of these aci-reductones , measured as a function of their inhibitory effect on CCl4-induced lipid peroxidation of hepatic microsomes , exceeds that produced by DB00163 . Synthetic routes and initial structure-activity relationships ( SAR ) for these novel mixed functioning antioxidants are presented . Alpha-tocopherol decreases superoxide anion release in human monocytes under hyperglycemic conditions via inhibition of protein kinase C-alpha . Diabetes is a major risk factor for premature atherosclerosis , and oxidative stress appears to be an important mechanism . Previously , we showed that diabetic monocytes produce increased superoxide anion ( O(2)(-) ) , and DB00163 ( AT ) supplementation decreases this . The aim of this study was to elucidate the mechanism(s) of O(2)(-) release and inhibition by AT under hyperglycemic ( HG ) conditions in monocytes . O(2)(-) release , protein kinase C ( PKC ) activity , and translocation of P17252 and -betaII and p47phox were increased in THP-1 cells ( human monocytic cell line ) under HG ( 15 mmol/l glucose ) conditions , whereas AT supplementation inhibited these changes . AT , NADPH oxidase inhibitors ( apocynin and diphenyleneiodonium chloride [ DPI ] ) , and an inhibitor to P17252 and other isoforms ( 2,2',3,3',4,4'-hexahydroxy-1,1'-biphenyl-6,6'-dimethanol dimethyl ether [ HBDDE ] ) but not P05771 II ( LY379196 ) decreased O(2)(-) release and p47phox translocation . Antisense oligodeoxynucleotides to P17252 and p47phox but not to PKC-betaII inhibited HG-induced O(2)(-) release and p47phox translocation in THP-1 cells . Under HG conditions , reactive oxygen species release from monocytes was not inhibited by agents affecting mitochondrial metabolism but was inhibited in human endothelial cells . We conclude that under HG conditions , monocytic O(2)(-) release is dependent on NADPH oxidase activity but not the mitochondrial respiratory chain ; HG-induced O(2)(-) release is triggered by P17252 , and AT inhibits O(2)(-) release via inhibition of P17252 . Anti-allergic function and regulatory mechanisms of KR62980 in allergen-induced airway inflammation . The ligand-activated transcription factor , peroxisome proliferator-activated receptor ( Q07869 )gamma , and its ligands inhibit pro-inflammatory cytokine production by immune cells , thus exerting anti-inflammatory activity . As a non-thiazolidinedione PPARgamma ligand , KR62980 has anti-diabetic and anti-adipogenic activities , but its anti-inflammatory function has yet to be characterized . In this study , we investigated the functions and mechanisms of KR62980 in the activation and differentiation of P01730 + T helper ( Th ) cells by comparing its effects with those of a thiazolidinedione PPARgamma ligand , rosiglitazone . KR62980 dose-dependently and significantly suppressed TCR-triggered Th cell proliferation by suppressing P60568 /IL-2Ralpha-mediated signaling . Both KR62980 and rosiglitazone suppressed IFNgamma production in a dose-dependent manner , whereas P05112 gene expression was specifically suppressed by only KR62980 . In addition , sustained KR62980 treatment diminished Th2 cytokine production by inhibiting c-Maf expression . In vivo administration of KR62980 in a model of allergic asthma significantly attenuated eotaxin-induced eosinophil infiltration , allergic cytokine production and collagen deposition in the lung . KR62980 also decreased goblet cell hyperplasia in the airway and mucous cell metaplasia in nasal epithelium , concurrent with decreases of allergic Th2 cytokines and Q16552 in the draining lymph node . In conclusion , a novel PPARgamma ligand , KR62980 , suppresses in vitro Th2 cell differentiation and attenuates in vivo OVA-induced airway inflammation , suggesting a beneficial role for KR62980 in the treatment of allergic asthma and allergic rhinitis . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Differential gene signatures in rat mammary tumors induced by DMBA and those induced by fractionated gamma radiation . The aim of this work was to identify specific genes involved in rat mammary tumors induced by dimethylbenz(a)anthracene ( DMBA ) or radiation . More TUNEL- and P12004 -positive cells were present in mammary tumors induced by radiation than in tumors induced by DMBA , whereas DNA damage responses like p53 accumulation and histone P16104 phosphorylation were higher in DMBA-induced tumors , even though the pathology was similar in both types of tumors . cDNA microarray and real-time RT-PCR analysis of radiation- or DMBA-induced tumor tissues , revealed that stanniocalcin 2 ( Stc2 ) , interferon regulatory factor 1 ( Irf1 ) , interleukin 18 binding protein ( Il18bp ) , and chloride channel calcium activated 3 ( Clca3 ) were expressed in both , and that arachidonate P09917 activating protein 1 ( Alox5ap ) and cathepsin S ( Ctss ) were expressed only in radiation-induced tumors . No DMBA-specific gene signatures were found . Soft agar growth assays were carried out to identify the carcinogenic features of these specific genes . Cells stably transfected with Alox5ap , Ctss , Stc2 , Irf1 , Il18bp and Clca3 showed morphological changes compared to controls . These findings indicate different gene alterations in carcinogen- or radiation-induced mammary tumors with similar pathological stages . ROS-mediated autophagy induced by dysregulation of lipid metabolism plays a protective role in colorectal cancer cells treated with gambogic acid . Gambogic acid ( GA ) , the main active component of gamboge resin , has potent antitumor activity both in vivo and in vitro . However , the underlying molecular mechanisms remain unclear . In this study , we found that GA could initiate autophagy in colorectal cancer cells , and inhibition of the autophagy process accelerated the effect of proliferative inhibition and apoptotic cell death induced by GA , implying a protective role of autophagy . Two-dimensional electrophoresis-based proteomics showed that GA treatment altered the expression of multiple proteins involved in redox signaling and lipid metabolism . Functional studies revealed that GA-induced dysregulation of lipid metabolism could activate P09917 ( 5- P28300 ) , resulting in intracellular ROS accumulation , followed by inhibition of Akt- P42345 signaling and autophagy initiation . Finally , results using a xenograft model suggested ROS-induced autophagy protect against the antitumor effect of GA . Taken together , these data showed new biological activities of GA against colorectal cancer underlying the protective role of ROS-induced autophagy . This study will provide valuable insights for future studies regarding the anticancer mechanisms of GA . TagSNP evaluation for the association of 42 inflammation loci and vascular disease : evidence of P05231 , P02675 , P09917 , P25963 , and P24394 loci effects . Inflammatory markers have consistently been associated with vascular disease . Evidence of genetic polymorphisms in inflammatory loci that predict severe carotid artery disease ( CAAD ) would suggest that this relationship is not secondary to other correlated factors , but related to inflammation itself . We examined the full common genetic variation in 42 inflammatory loci for prediction of severe CAAD versus ultrasound proven controls using a tagSNP approach . For selected loci , monocyte RNA levels were contrasted in subjects with and without CAAD . We confirm the association of P05231 (-174) , P02675 ( -455 ) , and P09917 with CAAD and show that multiple P09917 SNPs independently predict CAAD . We provide evidence for previously unreported associations of SNPs in P24394 , P25963 , and P00747 with CAAD , and weaker evidence for associations with P09919 , Q13651 , and P19320 . The P25963 and Q13651 expression levels significantly differed between subjects with CAAD and controls . These results support a role for genetic variation related to inflammation in CAAD and a causal role for specific gene products . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . DB01393 restores the inhibition of DB00094 -induced follicular development and steroidogenesis by tumor necrosis factor-alpha through peroxisome proliferator-activated receptor-gamma pathway in an in vitro mouse preantral follicle culture . We recently reported that bezafibrate , a lipid-lowering drug of the fibrate class , administered in addition to clomiphene citrate ( CC ) successfully induced ovulation in CC-resistant polycystic ovary syndrome ( PCOS ) patients . We hypothesized that bezafibrate may directly affect ovarian follicle development . P01308 resistance and compensatory hyperinsulinemia are important for the pathogenesis of PCOS . In this study , we first examined the effects of tumor necrosis factor-alpha ( P01375 ) , which plays a role in insulin resistance , on follicle development by using the follicle culture system . P01375 significantly inhibited follicle-stimulating hormone ( DB00094 ) -induced follicle development , 17beta-estradiol ( E2 ) secretion , and ovulation rate in a dose-dependent manner . We then examined whether bezafibrate treatment could rescue the inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 . DB01393 treatment rescued inhibition of follicle development , secretion of E2 , and ovulation rate by P01375 . We examined the expression of peroxisome proliferator-activated receptor ( Q07869 ) subtypes in mouse preantral follicles . As the protein expression of only P37231 was observed in mouse preantral follicles , we examined whether bezafibrate could affect follicle development and steroidogenesis through P37231 pathways . Treatment with GW1929 , a selective P37231 agonist , restored inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 , whereas treatment with GW9662 , a selective P37231 antagonist , canceled the restorative effects of bezafibrate . Collectively , the results in this study suggest that bezafibrate may directly exhibit a restorative effect on the inhibition of ovarian follicle development and steroidogenesis by P01375 through the P37231 pathway . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus ( FK506 ) . Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes ( EBV-CTLs ) to solid organ transplant ( SOT ) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders ( PTLDs ) . SOT recipients , however , require the continuous administration of immunosuppressive drugs to prevent graft rejection , and these agents may significantly limit the long-term persistence of transferred EBV-CTLs , precluding their use as prophylaxis . DB00864 ( FK506 ) is one of the most widely used immunosuppressive agents in SOT recipients , and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein ( P62942 ) . We have knocked down the expression of P62942 in EBV-CTLs using a specific small interfering RNA ( siRNA ) stably expressed from a retroviral vector and found that P62942 -silenced EBV-CTLs are FK506 resistant . These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity . We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model , suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation . Protein kinase C-alpha-mediated regulation of low-density lipoprotein receptor related protein and urokinase increases astrocytoma invasion . Aggressive and infiltrative invasion is one of the hallmarks of glioblastoma . P01130 -related protein ( Q14764 ) is expressed by glioblastoma , but the role of this receptor in astrocytic tumor invasion remains poorly understood . We show that activation of protein kinase C-alpha ( P17252 ) phosphorylated and down-regulated Q14764 expression . Pretreatment of tumor cells with PKC inhibitors , phosphoinositide 3-kinase ( PI3K ) inhibitor , P17252 small interfering RNA ( siRNA ) , and short hairpin RNA abrogated phorbol 12-myristate 13-acetate-induced down-regulation of Q14764 and inhibited astrocytic tumor invasion in vitro . In xenograft glioblastoma mouse model and in vitro transmembrane invasion assay , Q14764 -deficient cells , which secreted high levels of urokinase-type plasminogen activator ( uPA ) , invaded extensively the surrounding normal brain tissue , whereas the Q14764 -overexpressing and uPA-deficient cells did not invade into the surrounding normal brain . siRNA , targeted against uPA in Q14764 -deficient clones , attenuated their invasive potential . Taken together , our results strongly suggest the involvement of P17252 /PI3K signaling pathways in the regulation of Q14764 -mediated astrocytoma invasion . Thus , a strategy of combining small molecule inhibitors of P17252 and PI3K could provide a new treatment paradigm for glioblastomas . Adipose tissue endothelial cells from obese human subjects : differences among depots in angiogenic , metabolic , and inflammatory gene expression and cellular senescence . OBJECTIVE : Regional differences among adipose depots in capacities for fatty acid storage , susceptibility to hypoxia , and inflammation likely contribute to complications of obesity . We defined the properties of endothelial cells ( EC ) isolated from subcutaneous adipose tissue ( P21673 ) and visceral adipose tissue ( VAT ) biopsied in parallel from obese subjects . RESEARCH DESIGN AND METHODS : The architecture and properties of the fat tissue capillary network were analyzed using immunohistochemistry and flow cytometry . P28906 (+)/CD31(+) EC were isolated by immunoselection/depletion . Expression of chemokines , adhesion molecules , angiogenic factor receptors , as well as lipogenic and senescence-related genes were assayed by real-time PCR . Fat cell size and expression of hypoxia-dependent genes were determined in adipocytes from both fat depots . RESULTS : Hypoxia-related genes were more highly expressed in VAT than P21673 adipocytes . VAT adipocytes were smaller than P21673 adipocytes . Vascular density and EC abundance were higher in VAT . VAT-EC exhibited a marked angiogenic and inflammatory state with decreased expression of metabolism-related genes , including endothelial lipase , Q8IV16 , and Q07869 gamma . VAT-EC had enhanced expression of the cellular senescence markers , P17936 and γ- P16104 , and decreased expression of Q96EB6 . Exposure to VAT adipocytes caused more EC senescence-associated β-galactosidase activity than P21673 adipocytes , an effect reduced in the presence of vascular endothelial growth factor A ( P15692 ) neutralizing antibodies . CONCLUSIONS : VAT-EC exhibit a more marked angiogenic and proinflammatory state than P21673 -EC . This phenotype may be related to premature EC senescence . VAT-EC may contribute to hypoxia and inflammation in VAT . Alpha-tocopherol decreases tumor necrosis factor-alpha mRNA and protein from activated human monocytes by inhibition of P09917 . Cardiovascular disease is the leading cause of morbidity in Westernized populations . Low levels of DB00163 ( AT ) are associated with increased incidence of atherosclerosis and increased intakes appear to be protective . AT supplementation decreases interleukin 1 and 6 release from human monocytes . Thus , the aim of this study was to examine the effect of AT on an important proinflammatory cytokine , tumor necrosis factor-alpha ( P01375 ) release from human monocytes . AT supplementation ( 1200 IU/day for 3 months ) significantly decreased P01375 release from activated human monocytes . Mechanisms that were examined included its effect as a general antioxidant , its inhibitory effect on protein kinase C ( PKC ) , and the cycloxygenase-lipoxygenase pathway . While AT decreased P01375 release from activated monocytes , other antioxidants had no effect on P01375 release . Specific PKC inhibitors had no effect on P01375 release from activated monocytes . The inhibition of P01375 release by AT in activated monocytes was reversed by leukotriene B(4) ( Q06643 (4) ) , a major product of the P09917 ( P09917 ) pathway . Similar observations were seen with inhibitors of P09917 . Indomethacin , a P36551 inhibitor , in the presence and absence of AT failed to affect P01375 activity . These findings suggest that AT decreases P01375 release from activated human monocytes via inhibition of P09917 . Also , AT as well as a P09917 inhibitor significantly decreased P01375 mRNA . Furthermore , AT and the P09917 inhibitor decreased NFkappab-binding activity . Thus , in activated human monocytes , AT appears to inhibit P01375 mRNA and protein by inhibition of P09917 . DB00163 and drug metabolism . Tocopherols and tocotrienols are metabolized by side chain degradation initiated by cytochrome P450 ( CYP ) -catalyzed omega-hydroxylation followed by beta-oxidation . Whereas DB00163 is only poorly metabolized , high amounts of the final products , carboxyethyl hydroxychroman ( CEHC ) , are found from other tocols in HepG2 cells and in human urine . P08684 and P78329 were suggested to be involved in tocopherol degradation . P08684 metabolizes most of the drugs and is induced by many of its substrates via the activation of the pregnane X receptor ( O75469 ) . Also tocopherols and in particular tocotrienols induce the expression of a O75469 -driven reporter gene and the expression of endogenous P08684 and P20815 which is supported by sporadic publications spread over the last 30 years . The potential interference of vitamin E with drug metabolism is discussed in the light of related complications evoked by herbal remedies . Species difference in glucuronidation formation kinetics with a selective P42345 inhibitor . The mammalian target of rapamycin ( P42345 ) is a protein kinase that shows key involvement in age-related disease and promises to be a target for treatment of cancer . In the present study , the elimination of potent DB00171 -competitive P42345 inhibitor 3-(6-amino-2-methylpyrimidin-4-yl)-N-(1H-pyrazol-3-yl)imidazo[1,2-b]pyridazin-2-amine ( compound 1 ) is studied in bile duct-cannulated rats , and the metabolism of compound 1 in liver microsomes is compared across species . Compound 1 was shown to undergo extensive N-glucuronidation in bile duct-catheterized rats . N-glucuronides were detected on positions N1 ( M2 ) and N2 ( M1 ) of the pyrazole moiety as well as on the primary amine ( M3 ) . All three N-glucuronide metabolites were detected in liver microsomes of the rat , dog , and human , while primary amine glucuronidation was not detected in cynomolgus monkey . In addition , N1- and N2-glucuronidation showed strong species selectivity in vitro , with rat , dog , and human favoring N2-glucuronidation and monkey favoring N1-glucuronide formation . Formation of M1 in monkey liver microsomes also followed sigmoidal kinetics , singling out monkey as unique among the species with regard to compound 1 N-glucuronidation . In this respect , monkeys might not always be the best animal model for N-glucuronidation of uridine diphosphate glucuronosyltransferase ( P78381 ) 1A9 or P22309 substrates in humans . The impact of N-glucuronidation of compound 1 could be more pronounced in higher species such as monkey and human , leading to high clearance in these species . While compound 1 shows promise as a candidate for investigating the impact of pan- P42345 inhibition in vivo , opportunities may exist through medicinal chemistry efforts to reduce metabolic liability with the goal of improving systemic exposure . Protein kinase C activation and the development of diabetic complications . Recent studies have identified that the activation of protein kinase C ( PKC ) and increased diacylglycerol ( DAG ) levels initiated by hyperglycemia are associated with many vascular abnormalities in retinal , renal , and cardiovascular tissues . Among the various PKC isoforms , the beta- and delta-isoforms appear to be activated preferentially in the vasculatures of diabetic animals , although other PKC isoforms are also increased in the renal glomeruli and retina . The glucose-induced activation of PKC has been shown to increase the production of extracellular matrix and cytokines ; to enhance contractility , permeability , and vascular cell proliferation ; to induce the activation of cytosolic phospholipase A2 ; and to inhibit Na+-K+-ATPase . The synthesis and characterization of a specific inhibitor for P05771 isoforms have confirmed the role of PKC activation in mediating hyperglycemic effects on vascular cells , as described above , and provide in vivo evidence that PKC activation could be responsible for abnormal retinal and renal hemodynamics in diabetic animals . Transgenic mice overexpressing P05771 isoform in the myocardium developed cardiac hypertrophy and failure , further supporting the hypothesis that P05771 isoform activation can cause vascular dysfunctions . Interestingly , hyperglycemia-induced oxidative stress may also mediate the adverse effects of P05771 isoforms by the activation of the DAG-PKC pathway , since treatment with D- DB00163 was able to prevent many glucose-induced vascular dysfunctions and inhibit DAG-PKC activation . Clinical studies are now in progress to determine whether P05771 inhibition can prevent diabetic complications . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . Systems biology modeling of the radiation sensitivity network : a biomarker discovery platform . PURPOSE : The discovery of effective biomarkers is a fundamental goal of molecular medicine . Developing a systems-biology understanding of radiosensitivity can enhance our ability of identifying radiation-specific biomarkers . METHODS AND MATERIALS : Radiosensitivity , as represented by the survival fraction at 2 Gy was modeled in 48 human cancer cell lines . We applied a linear regression algorithm that integrates gene expression with biological variables , including ras status ( mut/wt ) , tissue of origin and p53 status ( mut/wt ) . RESULTS : The biomarker discovery platform is a network representation of the top 500 genes identified by linear regression analysis . This network was reduced to a 10-hub network that includes c-Jun , Q13547 , Q04206 ( p65 subunit of NFKB ) , P05771 , P63165 , c-Abl , P42224 , AR , P06493 , and P10914 . Nine targets associated with radiosensitization drugs are linked to the network , demonstrating clinical relevance . Furthermore , the model identified four significant radiosensitivity clusters of terms and genes . Ras was a dominant variable in the analysis , as was the tissue of origin , and their interaction with gene expression but not p53 . Overrepresented biological pathways differed between clusters but included DNA repair , cell cycle , apoptosis , and metabolism . The c-Jun network hub was validated using a knockdown approach in 8 human cell lines representing lung , colon , and breast cancers . CONCLUSION : We have developed a novel radiation-biomarker discovery platform using a systems biology modeling approach . We believe this platform will play a central role in the integration of biology into clinical radiation oncology practice . Intraretinal lipid transport is dependent on high density lipoprotein-like particles and class B scavenger receptors . PURPOSE : In our companion paper we demonstrated that circulating lipoproteins enter the retina via the retinal pigment epithelium ( Q96AT9 ) and possibly Müller cells . In order to understand how these lipids are transported within the retina , expression and localization of the main proteins known to be involved in systemic lipid transport was determined . METHODS : Expression of O95477 , apoA1 ( the major HDL protein ) , Q8WTV0 , SR- Q15878 , P16671 , lecithin:cholesterol acyltransferase ( P04180 ) , and cholesteryl ester transfer protein ( P11597 ) was determined by reverse transcriptase polymerase chain reaction ( RT-PCR ) and immunoblots . Localization was determined by immunohistochemistry using fresh monkey vibrotome sections and imaged by confocal microscopy . RESULTS : O95477 and apoA1 were localized to the ganglion cell layer , retinal pigment epithelium ( Q96AT9 ) , and rod photoreceptor inner segments . ApoA1 was also observed associated with rod photoreceptor outer segments , presumably localized to the interphotoreceptor matrix ( IPM ) . The scavenger receptors Q8WTV0 and SR- Q15878 localized mainly to the ganglion cell layer and photoreceptor outer segments ; in the latter they appear to be associated with microtubules . P04180 and P11597 localized mainly to the IPM . CONCLUSIONS : The presence and specific localization of these well-known lipid transport proteins suggest that the retina employs an internal lipid transport mechanism that involves processing and maturation of HDL-like particles . Niacin reduces plasma P11597 levels by diminishing liver macrophage content in P11597 transgenic mice . The anti-dyslipidemic drug niacin has recently been shown to reduce the hepatic expression and plasma levels of P11597 . Since liver macrophages contribute to hepatic P11597 expression , we investigated the role of macrophages in the P11597 -lowering effect of niacin in mice . In vitro studies showed that niacin does not directly attenuate P11597 expression in macrophages . Treatment of normolipidemic human P11597 transgenic mice , fed a Western-type diet with niacin for 4 weeks , significantly reduced the hepatic cholesterol concentration ( -20 % ) , hepatic P11597 gene expression ( -20 % ) , and plasma P11597 mass ( -30 % ) . Concomitantly , niacin decreased the hepatic expression of P34810 ( -44 % ) and P45844 ( -32 % ) , both of which are specific markers for the hepatic macrophage content . The decrease in hepatic P11597 expression was significantly correlated with the reduction of hepatic macrophage markers . Furthermore , niacin attenuated atherogenic diet-induced inflammation in liver , as evident from decreased expression of P01375 ( -43 % ) . Niacin similarly decreased the macrophage markers and absolute macrophage content in hyperlipidemic P02649 *3-Leiden. P11597 transgenic mice on a Western-type diet . In conclusion , niacin decreases hepatic P11597 expression and plasma P11597 mass by attenuating liver inflammation and macrophage content in response to its primary lipid-lowering effect , rather than by attenuating the macrophage P11597 expression level .

MH_train_1046

MH_train_1046

MH_train_1046

interacts_with DB06273?

multiple_choice

Adipose tissue dysfunction in nascent metabolic syndrome . The metabolic syndrome ( MetS ) confers an increased risk for both type 2 diabetes mellitus ( T2DM ) and cardiovascular disease ( CVD ) . Moreover , studies on adipose tissue biology in nascent MetS uncomplicated by T2DM and/or CVD are scanty . Recently , we demonstrated that adipose tissue dysregulation and aberrant adipokine secretion contribute towards the syndrome 's low-grade chronic proinflammatory state and insulin resistance . Specifically , we have made the novel observation that subcutaneous adipose tissue ( P21673 ) in subjects with nascent MetS has increased macrophage recruitment with cardinal crown-like structures . We have also shown that subjects with nascent MetS have increased the levels of P21673 -secreted adipokines ( IL-1 , P05231 , P10145 , leptin , P02753 -4 , CRP , P0DJI8 , P05121 , P13500 , and chemerin ) and plasma adipokines ( IL-1 , P05231 , leptin , P02753 -4 , CRP , P0DJI8 , and chemerin ) , as well as decreased levels of plasma adiponectin and both plasma and P21673 omentin-1 . The majority of these abnormalities persisted following correction for increased adiposity . Our data , as well as data from other investigators , thus , highlight the importance of subcutaneous adipose tissue dysfunction in subjects with MetS and its contribution to the proinflammatory state and insulin resistance . This adipokine profile may contribute to increased insulin resistance and low-grade inflammation , promoting the increased risk of T2DM and CVD . P00441 G93D sporadic amyotrophic lateral sclerosis ( SALS ) patient with rapid progression and concomitant novel P03950 variant . P00441 G93D mutation has been described in amyotrophic lateral sclerosis ( P35858 ) patients with slowly progressive disease . We describe an Italian patient affected by sporadic P35858 with the P00441 G93D mutation that disclosed an unusual rapid progression with death occurring after 30 months from the symptom onset . Considering the atypical clinical course further genes associated with P35858 or known to be causative were studied including P03950 , PGRN , Q13148 , P35637 , P55072 , P32297 , P43681 , and P30926 . A novel heterozygous P03950 missense variant ( c.433 C > T , p.R145C ) was identified which is neither reported in controls nor in 1000 genomes and single nucleotide polymorphism ( SNP ) databases . This report confirms that clinical course of P00441 -related P35858 may be modulated by other causative or associated genes , including P03950 and suggests that extensive screening of P35858 -associated genes in patients with an already identified mutation may be helpful for better knowledge of genetic architecture of P35858 . Modulation of Q8IVT5 activity in Caenorhabditis elegans by Zn ions , P25116 kinase and PP2A phosphatase . Vulval differentiation in Caenorhabditis elegans is controlled by a conserved signal transduction pathway mediated by Ras and a kinase cascade that includes Raf , Mek and MAPK . Activation of this cascade is positively regulated by a number of proteins such as Q8IVT5 ( kinase suppressor of Ras ) , Q09428 -8/ Q5T124 -2 , Q09428 -6/PP2A-B and P05231 -1 . We describe the functional characterization of sur-7 and several genes that regulate signaling downstream of ras . We identified sur-7 by isolating a mutation that suppresses an activated ras allele , and showed that Q09428 -7 is a divergent member of the cation diffusion facilitator family of heavy metal ion transporters that is probably localized to the endoplosmic recticulum membrane and regulates cellular Zn(2+) concentrations . Genetic double mutant analyses suggest that the Q09428 -7-mediated effect is not a general toxic response . Instead , Zn(2+) ions target a specific step of the pathway , probably regulation of the scaffolding protein Q8IVT5 . Biochemical analysis in mammalian cells indicates that high Zn(2+) concentration causes a dramatic increase of Q8IVT5 phosphorylation . Genetic analysis also indicates that PP2A phosphatase and P25116 kinase act downstream of Raf to positively and negatively regulate Q8IVT5 activity , respectively . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Genetic analysis of expression profile involved in retinoid metabolism in non-alcoholic fatty liver disease . AIM : The patients with non-alcoholic fatty liver disease ( NAFLD ) have been reported to be at greater risk for progression to chronic liver disease including liver cirrhosis ( LC ) . To examine the mechanisms for the progression of NAFLD , a genetic analysis of hepatic expression profile in retinoid metabolism in NAFLD was performed since the loss of retinoid signaling is associated with the progression of liver disease via reactive oxygen species ( ROS ) generation . METHODS : Fifty-one genes , which are associated with retinoid metabolism and action , were examined in thirty six subjects including 17 patients with simple steatosis , 11 with non-alcoholic steatohepatitis ( NASH ) and eight controls were examined by real-time reverse transcriptase polymerase chain reaction . Immunohistochemical study was also done by 3 kinds of antibodies . RESULTS : Higher expression of P09455 O95237 , DGT1/2 and CES1 in NAFLD suggests that mutual conversion between retinyl ester and retinal occurs actively . Expression of P07327 /2/3 , Q92781 /10/11 , O75911 and RALDH1/3 was increased in NAFLD , suggesting that oxidation process from retinol to all-trans retinoic acid ( DB00755 ) was enhanced . Importantly , greater expression of O43174 indicated that degradation of DB00755 was enhanced in NAFLD . Further , expression of P00441 /2 , catalase , thioredoxin and uncoupling protein 2 was also enhanced . CONCLUSION : Hyperdynamic state of retinoid metabolism is present in the liver tissues with NAFLD , which may be a putative mechanism by which NAFLD progresses to chronic liver disease including LC . Anti- P05231 -receptor-alpha ( tocilizumab ) does not inhibit human monocyte-derived dendritic cell maturation or alloreactive T-cell responses . Significant comorbidites and lethality complicate GVHD and its treatment . Targeting the cytokine milieu may improve GVHD control ; and P05231 is an attractive candidate , given its role in dendritic cell activation and T-cell differentiation . DB06273 is a humanized mAb to P05231 -receptor-α ( P08887 -α ) , which is Food and Drug Administration-approved for treatment of rheumatoid arthritis . Mouse transplant models have demonstrated that P05231 blockade also improves GVHD scores and survival . Definitive immunologic effects of P05231 inhibition have not emerged given inconsistent alterations in regulatory T cells ( Tregs ) and suppression of T-cell proliferation . Despite on-target suppression of P08887 -α signaling in human monocyte-derived dendritic cells ( moDCs ) and T cells , our data show no effect on moDC maturation/activation , alloreactive T-cell proliferation , Treg expansion , or allogeneic Th1/Th17 responses in vitro . These findings merit attention in any clinical trials of tocilizumab for GVHD prevention or treatment and provide a rationale for evaluating more specific inhibitors of downstream O60674 / P40763 signaling as well . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Temporal network based analysis of cell specific vein graft transcriptome defines key pathways and hub genes in implantation injury . Vein graft failure occurs between 1 and 6 months after implantation due to obstructive intimal hyperplasia , related in part to implantation injury . The cell-specific and temporal response of the transcriptome to vein graft implantation injury was determined by transcriptional profiling of laser capture microdissected endothelial cells ( EC ) and medial smooth muscle cells ( SMC ) from canine vein grafts , 2 hours ( H ) to 30 days ( D ) following surgery . Our results demonstrate a robust genomic response beginning at 2 H , peaking at 12-24 H , declining by 7 D , and resolving by 30 D . Gene ontology and pathway analyses of differentially expressed genes indicated that implantation injury affects inflammatory and immune responses , apoptosis , mitosis , and extracellular matrix reorganization in both cell types . Through backpropagation an integrated network was built , starting with genes differentially expressed at 30 D , followed by adding upstream interactive genes from each prior time-point . This identified significant enrichment of P05231 , P10145 , NF-κB , dendritic cell maturation , glucocorticoid receptor , and Triggering Receptor Expressed on Myeloid Cells ( Q9NP99 ) signaling , as well as PPARα activation pathways in graft EC and SMC . Interactive network-based analyses identified P05231 , P10145 , IL-1α , and P01308 Receptor ( P06213 ) as focus hub genes within these pathways . Real-time PCR was used for the validation of two of these genes : P05231 and P10145 , in addition to Collagen 11A1 ( P12107 ) , a cornerstone of the backpropagation . In conclusion , these results establish causality relationships clarifying the pathogenesis of vein graft implantation injury , and identifying novel targets for its prevention . DB06273 in pediatric rheumatology : the clinical experience . During the last two decades , clinical use of novel biological therapy has led to increased mechanistic understanding of complex rheumatological diseases . Conversely , basic and translational studies have led to development of new and varied therapeutic agents . These new medications which " target " specific steps in one or more immune pathways have the potential to control disease symptoms , improve quality of life and long-term prognosis , and perhaps in some , restore immunological tolerance . Use of these agents in clinical trials , combined with post-marketing surveillance , has revealed both the benefits and the undesirable side-effects of biological disease-modifying anti-rheumatic drugs ( DMARDs ) . In this review we focus on the use of tocilizumab , a monoclonal antibody directed against the P05231 receptor ( P08887 ) , which potently inhibits P05231 / P08887 signaling . Different cholinesterase inhibitor effects on P04141 cholinesterases in Alzheimer patients . BACKGROUND : The current study aimed to compare the effects of different cholinesterase inhibitors on acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) activities and protein levels , in the cerebrospinal fluid ( P04141 ) of Alzheimer disease ( AD ) patients . METHODS AND FINDINGS : AD patients aged 50-85 years were randomized to open-label treatment with oral rivastigmine , donepezil or galantamine for 13 weeks . P22303 and BuChE activities were assayed by Ellman 's colorimetric method . Protein levels were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Primary analyses were based on the Completer population ( randomized patients who completed Week 13 assessments ) . 63 patients were randomized to treatment . DB00989 was associated with decreased P22303 activity by 42.6 % and decreased P22303 protein levels by 9.3 % , and decreased BuChE activity by 45.6 % and decreased BuChE protein levels by 21.8 % . DB00674 decreased P22303 activity by 2.1 % and BuChE activity by 0.5 % , but increased P22303 protein levels by 51.2 % and BuChE protein levels by 10.5 % . Donepezil increased P22303 and BuChE activities by 11.8 % and 2.8 % , respectively . Donepezil caused a 215.2 % increase in P22303 and 0.4 % increase in BuChE protein levels . Changes in mean P22303 -Readthrough/Synaptic ratios , which might reflect underlying neurodegenerative processes , were 1.4 , 0.6 , and 0.4 for rivastigmine , donepezil and galantamine , respectively . CONCLUSION : The findings suggest pharmacologically-induced differences between rivastigmine , donepezil and galantamine . DB00989 provides sustained inhibition of P22303 and BuChE , while donepezil and galantamine do not inhibit BuChE and are associated with increases in P04141 P22303 protein levels . The clinical implications require evaluation . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . DB00184 consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 - P30532 - P30926 cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5(-/-) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5*-nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake . Pituitary-adrenal axis regulation in P06850 -deficient mice . P06850 ( P06850 ) -deficient ( knockout ( KO ) ) mice demonstrate severely impaired adrenal responses to restraint , ether , and fasting , and lack the normal diurnal glucocorticoid ( GC ) rhythm . Here , we summarize recent studies determining the role of P06850 in augmenting plasma adrenocorticotrophic hormone ( DB01285 ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 ) mRNA , DB01285 peptide content within the pituitary , and plasma DB01285 concentrations are not elevated in P06850 KO mice . P01189 mRNA content in P06850 KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 mRNA , plasma DB01285 does not increase in the P06850 KO mice following adrenalectomy . Administration of P06850 to adrenalectomized P06850 KO mice results in acute , robust DB01285 secretion . Thus , loss of GC feedback can increase P01189 gene expression in the pituitary , but P06850 action is essential for increased secretion of DB01285 into the circulation . While GC secretion is impaired in P06850 KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 and P05231 deficiency reveal that P05231 is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Interferon-gamma-induced dephosphorylation of P40763 and apoptosis are dependent on the P42345 pathway . Interferon-gamma ( P01579 ) exhibits diverse biological activities , including control of cell growth and tumor suppression . Here , we report that the treatment of M12 cells , a human metastatic prostate cancer cell line , with P01579 , resulted in marked inhibition of cell proliferation and induced apoptosis . These effects were not seen with either IFN-alpha or IFN-beta . M12 cells , like many other human cancer cells , contain constitutively activated signal transducer and activator of transcription 3 ( P40763 ) . The basal levels of both Akt and P27361 /2 phosphorylation are also markedly elevated in M12 cells . Strikingly , P01579 -induced apoptosis and growth inhibition of M12 cells were associated with persistent suppression of the constitutive tyrosine-phosphorylated P40763 ( pY- P40763 ) . The P01579 -induced dephosphorylation of pY- P40763 , however , was inhibited when the P42345 pathway was specifically blocked by rapamycin . Inhibition of PI-3K with low-dose LY294002 , or MAPK with PD98059 also suppressed the P42345 / P08133 S6k pathway , and correlated with the blockage of P01579 -induced dephosphorylation of pY- P40763 . Simultaneously , treatment with LY294002 , PD98059 , or rapamycin abolished P01579 -induced apoptosis in M12 cells . The inhibition of the P42345 pathway , however , did not affect P01579 -induced activation of P42224 pathway , and suppression of P42224 expression by siRNA had no effect on P01579 -induced dephosphorylation of pY- P40763 . Taken together , these results demonstrate that an intact P42345 pathway is critical for P01579 -induced suppression of pY- P40763 and apoptosis . Our study thus provides novel insights into the contributions of signaling pathways other than the classical JAK/ P42224 pathway in the anti-proliferative , proapoptotic actions of P01579 . Signaling by retinol and its serum binding protein . DB00162 , retinol , circulates in blood bound to retinol-binding protein ( P02753 ) which , in turn , associates with transthyretin ( P02766 ) to form a retinol- P02753 - P02766 ternary complex . At some tissues , retinol-bound ( holo- ) P02753 is recognized by a membrane protein termed Q9BX79 , which transports retinol from extracellular P02753 into cells and , concomitantly , activates a O60674 / P40763 /5 signaling cascade that culminates in induction of P35610 target genes . Q9BX79 -mediated retinol transport and cell signaling are critically inter-dependent , and they both require the presence of cellular retinol-binding protein 1 ( P09455 ) , an intracellular retinol acceptor , as well as a retinol-metabolizing enzyme such as lecithin:retinol acyltransferase ( O95237 ) . Q9BX79 thus functions as a " cytokine signaling transporter " which couples vitamin A homeostasis and metabolism to cell signaling , thereby regulating gene transcription . Recent studies provided molecular level insights into the mode of action of this unique protein . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor . The aryl hydrocarbon receptor ( P35869 ) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways , such as xenobiotic metabolism and Th17 cell differentiation . Identification of key physiologically relevant ligands that regulate P35869 function remains to be accomplished . Screening of indole metabolites has identified indoxyl 3-sulfate ( I3S ) as a potent endogenous ligand that selectively activates the human P35869 at nanomolar concentrations in primary human hepatocytes , regulating transcription of multiple genes , including P04798 , P05177 , Q16678 , P22309 , P19224 , P05231 , and P0DJI8 . Furthermore , I3S exhibits an approximately 500-fold greater potency in terms of transcriptional activation of the human P35869 relative to the mouse P35869 in cell lines . Structure-function studies reveal that the sulfate group is an important determinant for efficient P35869 activation . This is the first phase II enzymatic product identified that can significantly activate the P35869 , and ligand competition binding assays indicate that I3S is a direct P35869 ligand . I3S failed to activate either CAR or O75469 . The physiological importance of I3S lies in the fact that it is a key uremic toxin that accumulates to high micromolar concentrations in kidney dialysis patients , but its mechanism of action is unknown . I3S represents the first identified relatively high potency endogenous P35869 ligand that plays a key role in human disease progression . These studies provide evidence that the production of I3S can lead to P35869 activation and altered drug metabolism . Our results also suggest that prolonged activation of the P35869 by I3S may contribute to toxicity observed in kidney dialysis patients and thus represent a possible therapeutic target . Expression of cytosolic retinoid-binding protein genes in human skin biopsies and cultured keratinocytes and fibroblasts . Using reverse transcription coupled to polymerase chain reaction we have studied the mRNA expression of serum retinol-binding protein and cytosolic receptors for retinol and retinoic acid in skin biopsies , and in cultured epidermal keratinocytes and dermal fibroblasts . Transcripts for cellular retinol-binding protein ( P09455 ) I and cellular retinoic-acid-binding protein ( CRABP ) I were found in normal skin , keratinocytes , and fibroblasts . CRABP II transcripts were detected in skin and keratinocytes . A decreased mRNA expression of CRABP I and an increased mRNA expression of CRABP II were found in lesional psoriatic skin compared with uninvolved skin . mRNA transcripts for serum retinol-binding protein ( s- P02753 ) were detected in all tissues and cells . The biological importance of s- P02753 expression in keratinocytes and fibroblasts is not known , but hypothetically this protein may be involved in the intracellular shuttling of retinol and retinoic acid , or in the retransportation of cellular retinoids into the extracellular space . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . P05231 -receptor polymorphisms rs12083537 , rs2228145 , and rs4329505 as predictors of response to tocilizumab in rheumatoid arthritis . DB06273 ( TCZ ) , a monoclonal antibody targeting the human interleukin-6-receptor ( IL-6R ) , is indicated for the treatment of rheumatoid arthritis ( RA ) . We examined whether three P08887 single-nucleotide polymorphisms rs12083537 , rs2228145 ( formerly rs8192284 ) , and rs4329505 with previously reported functional effects were associated with clinical response to TCZ in a retrospective study cohort consisting of 79 RA patients . Three months after initiation of TCZ therapy , changes in swollen joint count ( SJC ) and , subordinately , tender joint count ( TJC ) , serum-CRP , DAS28-CRP , and EULAR-response were tested for association with the P08887 -haplotype or genotype . The major allele ( A ) of rs12083537 and the minor allele ( C ) of rs4329505 were associated with a poor SJC response ( P=0.02 and 0.02 , respectively ) . Moreover , the AAC-haplotype ( for rs12083537 , rs2228145 , and rs4329505 , respectively ) was associated with a poor SJC response ( P=0.00004 ) and , with borderline significance , EULAR-response ( P=0.05 ) . These data suggest that genetic variation in P08887 may aid in predicting TCZ therapy outcome in RA patients .

MH_train_1047

MH_train_1047

MH_train_1047

interacts_with DB00624?

multiple_choice

The androgen axis in recurrent prostate cancer . PURPOSE : Prostate cancer that recurs during androgen deprivation therapy is referred to as androgen-independent . High levels of expression of androgen receptor and androgen receptor-regulated genes in recurrent prostate cancer suggest a role for androgen receptor and its ligands in prostate cancer recurrence . EXPERIMENTAL DESIGN : Recurrent prostate cancer specimens from 22 men whose prostate cancer recurred locally during androgen deprivation therapy and benign prostate specimens from 48 men who had received no prior treatment were studied . P10275 expression was measured using monoclonal antibody and automated digital video image analysis . Tissue androgens were measured using radioimmunoassay . RESULTS : Epithelial nuclei androgen receptor immunostaining in recurrent prostate cancer ( mean optical density , 0.284 +/- SD 0.115 and percentage positive nuclei , 83.7 +/- 11.6 ) was similar to benign prostate ( mean optical density , 0.315 +/- 0.044 and percentage positive nuclei , 77.3 +/- 13.0 ) . Tissue levels of testosterone were similar in recurrent prostate cancer ( 2.78 +/- 2.34 pmol/g tissue ) and benign prostate ( 3.26 +/- 2.66 pmol/g tissue ) . Tissue levels of dihydrotestosterone , dehydroepiandrosterone , and androstenedione were lower ( Wilcoxon , P = 0.0000068 , 0.00093 , and 0.0089 , respectively ) in recurrent prostate cancer than in benign prostate , and mean dihydrotestosterone levels , although reduced , remained 1.45 nM . P10275 activation in recurrent prostate cancer was suggested by the androgen-regulated gene product , prostate-specific antigen , at 8.80 +/- 10.80 nmol/g tissue . CONCLUSIONS : DB00624 and dihydrotestosterone occur in recurrent prostate cancer tissue at levels sufficient to activate androgen receptor . Novel therapies for recurrent prostate cancer should target androgen receptor directly and prevent the formation of androgens within prostate cancer tissue . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . The androgen receptor : a mediator of diverse responses . Androgens mediate a number of diverse responses through the androgen receptor , a 110 kD ligand-activated nuclear receptor . P10275 expression , which is found in a variety of tissues , changes throughout development , aging , and malignant transformation . The androgen receptor can be activated by two ligands , testosterone and dihydrotestosterone , which bind to the androgen receptor with different affinities . This difference in binding affinity results in different levels of activation of the androgen receptor by the two ligands . The androgen receptor acts as a transcriptional modifier of a variety of genes by binding to an androgen response element . The ability to confer androgen specific actions by the androgen response element may depend on other cell-specific transcription factors and cis-acting DNA elements in close proximity to it . DB00624 and dihydrotestosterone appear to act upon an identical nuclear receptor . However , in certain instances , they mediate different physiologic responses . For example , dihydrotestosterone , but not testosterone , is capable of mediating full sexual development of the male external genitalia . In some cases , the androgen receptor may induce opposite physiologic responses in similar tissue types depending on their location . For example , in male pattern baldness , activated androgen receptors may suppress the growth of distinct hair follicle populations through initiating stromal-epithelial actions , whereas other hair follicles continue to proliferate . In other cases , altered androgen receptor activity due to its mutation or altered expression may lead to pathology such as recurrence of prostate cancer due to development of androgen independence allowing tumor cell proliferation under androgen deprivation . Inhibition of tumor growth and progression of LNCaP prostate cancer cells in athymic mice by androgen and liver X receptor agonist . Androgen-dependent human LNCaP 104-S tumor xenografts progressed to androgen-independent relapsed tumors ( 104-Rrel ) in athymic mice after castration . The growth of 104-Rrel tumors was suppressed by testosterone . However , 104-Rrel tumors adapted to androgen and regrew as androgen-stimulated 104-Radp tumors . P10275 expression in tumors and serum prostate-specific antigen increased during progression from 104-S to 104-Rrel but decreased during transition from 104-Rrel to 104-Radp . Expression of genes related to liver X receptor ( LXR ) signaling changed during progression . LXRalpha , LXRbeta , DB00171 -binding cassette transporter A1 ( O95477 ) , and sterol 27-hydroxylase decreased during progression from 104-S to 104-Rrel . These coordinated changes in LXR signaling in mice during progression are consistent with our previous findings that reduction of O95477 gene expression stimulates proliferation of LNCaP cells . To test if attenuation of LXR signaling may enhance prostate cancer progression from an androgen-dependent state to an androgen-independent state , castrated mice carrying 104-S tumors were given the synthetic LXR agonist T0901317 by gavage . T0901317 delayed progression from 104-S to 104-Rrel tumors . Based on our in vivo model , androgen is beneficial for the treatment of androgen-independent androgen receptor-rich prostate cancer and modulation of LXR signaling may be a potentially useful therapy for prostate cancer . P10275 signaling induced by supraphysiological doses of dihydrotestosterone in human peripheral blood lymphocytes . Anabolic androgenic steroids , a class of steroid hormones related to testosterone , are natural ligands of androgen receptor ( AR ) , a member of the nuclear receptor superfamily of ligand-activated transcription factors . AR binds specific DNA elements , known as androgen-response elements . DB00624 , the main male sexual hormone , binds AR directly and indirectly , through conversion into dihydrotestosterone ( DB02901 ) , its more active metabolite . Anabolic androgenic steroids are frequently detected in the urine of doped athletes ; their consumption is also growing among sport amateurs and adolescents . The effects of androgens can differ depending on the target cells and/or tissues . To gain insight into transcription activation mechanisms of AR , we investigated AR protein signaling in human peripheral blood lymphocytes treated with supraphysiological doses of DB02901 . We performed a comparative proteomic analysis and we identified about 30 differentially expressed proteins . At least five species contained a consensus androgen-response elements sequence in the promoter region of related coding genes . The analysis also revealed that high doses of DB02901 activate the drug detoxification process , could stimulate an increase in cell motility and exert a prosurvival effect rather than an apoptotic one . P10275 -dependent transactivation of growth arrest-specific gene 6 mediates inhibitory effects of testosterone on vascular calcification . Recent epidemiological studies have found that androgen deficiency is associated with a higher incidence of cardiovascular disease in men . However , little is known about the mechanism underlying the cardioprotective effects of androgens . Here we show the inhibitory effects of testosterone on vascular calcification and a critical role of androgen receptor ( AR ) -dependent transactivation of growth arrest-specific gene 6 ( Gas6 ) , a key regulator of inorganic phosphate ( P(i) ) -induced calcification of vascular smooth muscle cells ( VSMC ) . DB00624 and nonaromatizable androgen dihydrotestosterone inhibited P(i)-induced calcification of human aortic VSMC in a concentration-dependent manner . Androgen inhibited P(i)-induced VSMC apoptosis , an essential process for VSMC calcification . The effects on VSMC calcification were mediated by restoration of P(i)-induced down-regulation of Gas6 expression and a subsequent reduction of Akt phosphorylation . These effects of androgen were blocked by an AR antagonist , flutamide , but not by an estrogen receptor antagonist , DB00947 . We then explored the mechanistic role of the AR in Gas6 expression and found an abundant expression of AR predominantly in the nucleus of VSMC and two consensus ARE sequences in the Gas6 promoter region . DB02901 stimulated Gas6 promoter activity , and this effect was abrogated by flutamide and by AR siRNA . Site-specific mutation revealed that the proximal ARE was essential for androgen-dependent transactivation of Gas6 . Furthermore , chromatin immunoprecipitation assays demonstrated ligand-dependent binding of the AR to the proximal ARE of Gas6 . These results indicate that AR signaling directly regulates Gas6 transcription , which leads to inhibition of vascular calcification , and provides a mechanistic insight into the cardioprotective action of androgens . P10275 expressing neurons that project to the paraventricular nucleus of the hypothalamus in the male rat . Androgen receptors are distributed throughout the central nervous system and are contained by a variety of nuclei that are known to project to or regulate the paraventricular nucleus ( PVN ) of the hypothalamus , the final common pathway by which the brain regulates the hypothalamic-pituitary-adrenal ( Q9Y251 ) response to homeostatic threat . Here we characterized androgen receptor staining within cells identified as projecting to the PVN in male rats bearing iontophoretic or crystalline injections of the retrograde tracer FluoroGold aimed at the caudal two-thirds of the nucleus , where corticotropin-releasing hormone-expressing neurons are amassed . P10275 ( AR ) and FluoroGold ( FG ) double labeling was revealed throughout the limbic forebrain , including scattered numbers of cells within the anterior and posterior subdivisions of the bed nuclei of the stria terminalis ; the medial zone of the hypothalamus , including large numbers of AR-FG-positive cells within the anteroventral periventricular and medial preoptic cell groups . Strong and consistent colabeling was also revealed throughout the hindbrain , predominantly within the periaqueductal gray and the lateral parabrachial nucleus , and within various medullary cell groups identified as catecholaminergic , predominantly C1 and A1 neurons of the ventral medulla . These connectional data predict that androgens can act on a large assortment of multimodal inputs to the PVN , including those involved with the processing of various types of sensory and limbic information , and provide an anatomical framework for understanding how gonadal status could contribute to individual differences in Q9Y251 function . P10275 -immunoreactive cells in ram hypothalamus : distribution and co-localization patterns with gonadotropin-releasing hormone , somatostatin and tyrosine hydroxylase . DB00624 exerts important feedback effects on the hypothalamus of the ram to influence reproductive functioning . To provide a neuroanatomical basis for understanding this androgen action , the present study has examined androgen receptor ( AR ) immunoreactivity within the hypothalamus and adjacent brain areas of the intact non-breeding season ram . The largest populations of AR-immunoreactive cells were detected in the medial preoptic area , infundibular and premammillary nuclei in addition to the ventromedial nucleus ( VMN ) where cells were found distributed throughout its medial and lateral divisions . Smaller numbers of AR-expressing cells were identified in the bed nucleus of the stria terminalis and anterior hypothalamic area ( DB00551 ) including the paraventricular , but not the supraoptic , nucleus . Double-labelling immunocytochemistry revealed the presence of AR immunoreactivity in only 2 of 460 gonadotropin-releasing hormone ( DB00644 ) neurons . A very small population of TH-immunoreactive cells located in the lateral aspect of the DB00551 was found to contain ARs . Dopaminergic cells elsewhere in the hypothalamus , including the infundibular nucleus , did not display AR immunoreactivity . Nearly 50 % of AR-expressing cells in the lateral VMN were immunoreactive for somatostatin while less than 5 % of periventricular somatostatin neurons displayed AR immunoreactivity . These results show where ARs are expressed in the ram hypothalamus and indicate the neuroanatomical sites at which androgen may act to influence reproductive function . The absence of ARs in the neuroendocrine DB00644 and tuberoinfundibular dopaminergic cells suggests that androgens do not influence the genome of these cells in any direct manner . In contrast , the somatostatin neurons of the VMN appear to be an important target for circulating androgens in the non-breeding season ram . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . Gene expression in sexually dimorphic muscles in sheep . DB00624 is known to act differentially on skeletal muscle from different regions of the body . Two genes likely to mediate the testosterone effect are insulin-like growth factor I ( P05019 ) , an important growth regulator acting in an autocrine and paracrine way , and androgen receptor ( AR ) , because receptor density could account for differential muscle growth . Another muscle-specific gene that may play a role in differential muscle growth is myostatin , a member of the transforming growth factor-beta superfamily , shown to be a negative regulator of skeletal muscle mass . The objective of this study was to quantify and compare the steady state expression of these three genes in two different skeletal muscles in sheep . Eleven Dorset rams were slaughtered after reaching puberty and total RNA was extracted from samples of semitendinosus and splenius muscles . P05019 mRNA was measured using a competitive reverse-transcription-polymerase chain reaction . P10275 and myostatin mRNA were measured by a ribonuclease protection assay ( RPA ) with standard curves . The means ( attomoles/microg RNA ) for splenius and semitendinosus muscles were 1.39 and 1.02 ( SE = 0.14 ) , 4.05 and 2.96 ( SE = 0.24 ) , and 4.30 and 3.85 ( SE = 0.37 ) for P05019 , AR , and myostatin , respectively . The difference between the two muscles was significant for P05019 and AR mRNA levels with higher levels in the splenius but not significant for myostatin . Our results show that locally produced P05019 and the regulation of AR expression may be important for sexually dimorphic muscle growth patterns . Androgen receptors . DB00624 and its active metabolite dihydrotestosterone exert their influence on target cells through a specific intracellular protein receptor . Structural abnormalities of this receptor lead to a diminished androgen action within the cell and result in the syndrome of androgen insensitivity . Androgen insensitivity is classified on the basis of whether the insensitivity is complete or partial and whether the androgen receptor is normally present ( AR(+) ) , absent ( AR(-) ) or diminished ( AR(+/-) ) . All patients with androgen insensitivity have normal or high plasma levels of testosterone and elevated serum LH . Patients with complete androgen insensitivity are phenotypically female . The clinical presentation of partial androgen insensitivity is variable , ranging from a minimal amount of virilization to a completely masculine appearance . All patients described with a syndrome of androgen insensitivity are infertile . The influence of androgen receptor function in the pathogenesis of benign prostatic hypertrophy is being investigated . P10275 content is also being studied as a possible marker of responsiveness to hormonal therapy in prostatic carcinoma . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . P10275 is widely expressed in bovine placentomes and up-regulated during differentiation of bovine trophoblast giant cells . OBJECTIVES : In cattle no biological role has been definitely identified for placental estrogens and progesterone . However , in the bovine trophoblast androgens may also be produced and have local effects . Thus , the aims of this study were to identify androgen receptor ( AR ) expressing cells and to monitor testosterone tissue concentrations in bovine placentomes throughout gestation . METHODS : Placental AR expression was characterized at the mRNA and protein level applying conventional and real-time RT-qPCR , western blot and immunohistochemistry . DB00624 was measured by radioimmunoassay . RESULTS : AR-mRNA was qualitatively detected from day 50 of gestation until term . Mean relative gene expression levels were constant between day 100 and late gestation . A slight non-significant increase was observed in the prepartal period . With immunohistochemistry distinct nuclear signals were predominantly observed in invasive trophoblast giant cells ( P21980 ) from day 80 until term . In mature P21980 of the trophoblast , immature P21980 and uninucleated trophoblast cells , stromal cells of the chorionic villi , caruncular epithelial and stromal cells immunoreactive score values were low at early and midgestation but increased significantly ( p < 0.01 ) during late gestation and remained high until parturition . With western blot in placentomal tissue a specific band of approximately 110 kDa was detected as it was the case in epididymis used as a positive control . DB00624 concentrations increased from 0.70 ± 0.29 pmol/g wet tissue between days 60-220 to 4.22 ± 1.29 pmol/g during late gestation ( p < 0.001 ) . DISCUSSION : The results are consistent with androgens as active products of bovine placental steroidogenesis . The substantial up-regulation of AR expression during P21980 differentiation suggests that androgens may be related to this process . P10275 in nuclei of rat testis . Testis nuclei of hypophysectomized rats selectively accumulate labeled testosterone and 5alpha-dihydrotestosterone following the injection of tritiated testosterone in vivo . DB00624 and 5alpha-dihydrotestosterone are bound to macromolecules in nuclei and can be extracted with 0.5 M DB00761 . Accumulation of protein bound radioactive androgens in nuclei of isolated seminiferous tubules is similar to that of whole testis . The relative amounts of testosterone and dihydrotestosterone in purified nuclei were similar to the relative amounts bound to cytoplasmic receptors , suggesting that cytoplasmic androgen-receptor complexes may be transported into the nuclei . Binding of labeled androgen is saturable and inhibited by prior injection of unlabeled testosterone or cyproterone acetate . Nuclear binding sites are destroyed by the proteolytic enzyme pronase , but not by DNase . Like the cytoplasmic androgen-receptor complexes in rat testis , nuclear androgen-protein complexes are heat labile and dissociate slowly at 0 degrees C. androgens fail to accumulate in testis nuclei of the Stanley-Gumbreck androgen insensitive rat , a species lacking cytoplasmic androgen receptors in testis and other androgen target tissues . P10275 is responsible for rat organic cation transporter 2 gene regulation but not for rOCT1 and rOCT3 . PURPOSE : Organic cation transporters 1-3 ( OCT1-3 ; Slc22a1-3 ) mediate the membrane transport of organic cations in the kidney . We previously reported that rat (r)OCT2 expression in the kidney was regulated by testosterone . In this study , we examined the transcriptional mechanisms underlying the testosterone-dependent regulation of rOCT2 expression . METHODS : Approximately 3000-bp fragments of the rOCT1-3 promoter region were isolated , and promoter activities were measured in the renal epithelial cell line LLC- P30613 with the coexpression of rat androgen receptor . RESULTS : Among reporter constructs tested , only rOCT2 promoter activity was stimulated by testosterone . This stimulation was suppressed by nilutamide , an antiandrogen drug . Reporter assays using deletion constructs and mutational constructs of putative androgen response elements ( ARE ) in the rOCT2 promoter region suggested that two AREs , located at approximately -3000 and -1300 , respectively , play an important role in the induction by testosterone . CONCLUSIONS : DB00624 induces the expression of rOCT2 , but not of rOCT1 and rOCT3 , via the AR-mediated transcriptional pathway . This is the first study to address the transcriptional mechanisms of testosterone-dependent gene regulation of the Slc22 family . P10275 in human skin cytosol . Human skin , an accessible tissue , is an androgen target organ . We have measured the androgen-binding capacity of human skin cytosol using either 5 alpha[3H]dihydrotestosterone ( [3H] DB02901 ) or [3H]methyltrienolone ( [3H]R-1881 ) as ligand . Samples were incubated for 20 h at 0 C , and dextran-coated charcoal was used to separate bound from free steroids . The androgen receptor has a high affinity for both ligands ( 0.23 +/- 0.04 nM for [3H] DB02901 ; 0.32 +/- 0.16 nM for [3H]R-1881 ) . DB00624 , cyproterone acetate , and , to a lesser extent , estradiol also bind this protein . Progesterone displaces R-1881 from its binding sites , whereas its 5 alpha-reduced metabolite somewhat inhibits DB02901 binding . The highest binding capacity is measured in cytosol of skin from external genitalia ( 129.14 +/- 58.0 fmol/g skin ; n = 34 ) ; it is lower in pubic skin ( 21.8 +/- 13 fmol/g skin ; n = 6 ) . There is no variation as a function of age or sex in genital skin ; the higher concentrations observed in the cytosol of pubic skin of women compared to that of men are probably related to lower levels of endogenous steroids . Whereas most patients with the complete form of the testicular feminization syndrome do not have detectable concentrations of androgen receptor , one patient with apparent complete clinical androgen insensitivity had a normal androgen-binding capacity . The parity of values in genital skin from men and women , the absence of variation with age , and the presence of a cytosolic androgen receptor in some androgen-insensitive patients suggest that the androgen receptor in human skin cytosol is not regulated by androgens . P10275 regulation of P55008 cyclin and cyclin-dependent kinase function in the CWR22 human prostate cancer xenograft . Human prostate cancer is initially dependent on androgens for growth , and androgen-dependent cells undergo apoptosis after castration . However , a subset of androgen-responsive cells survives and eventually proliferates in the absence of testicular androgen . The high levels of androgen receptor in both androgen-dependent and recurrent tumors led us to investigate androgen regulation of cell cycle proteins in human prostate cancer using the CWR22 xenograft . Cellular proliferation decreased dramatically in CWR22 tumors after castration . DB00624 propionate ( TP ) treatment of castrated mice restored cellular proliferation after 24-48 hours . Growth of CWR22 tumors in the absence of testicular androgen recurred several months after castration . P06493 and P24941 , and cyclin A and cyclin B1 messenger RNAs were decreased 6 days after castration , increased 6-12 hours after TP treatment , and were expressed at high levels in recurrent CWR22 tumors . Coimmunoprecipitated cyclin B1/ P06493 and cyclin D1/ P11802 protein complexes decreased after castration and increased after TP treatment of castrated mice . In addition , P06493 and P24941 kinase activities were upregulated by androgen in parallel with hyperphosphorylation of retinoblastoma ( Rb ) protein . Despite the absence of testicular androgen in recurrent CWR22 , the levels of these androgen-regulated cyclin/ CDK protein complexes and hyperphosphorylation of Rb were equal to or greater than in tumors from intact mice . The results indicate that androgen receptor regulates cellular proliferation by control of CDK and cyclins at the transcriptional level and by post-translational modifications that influence cell cycle protein activity . Recent androgen receptor antagonists in prostate cancer . P10275 has been shown to promote prostate cell growth and carcinogenesis of prostate cancer by up-regulating its target genes . DB00624 and dihydrotestosterone are two major hormones which bind to and activate androgen receptor . Targeting both the androgen receptor and the enzymes catalyzing the biosynthesis of testosterone and dihydrotestosterone has been shown to be clinically beneficial in the treatment of prostate cancer . Prostate cancer can become castration-resistant after long term treatment with chemo drugs , so efforts in finding compounds with improved efficiency to castration-resistant prostate cancer are urgently needed . In this review we summarized the studies on recent progress in the development of small molecular AR antagonists for the treatment of prostate cancer . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . Gender-specific effects of endogenous testosterone : female alpha-estrogen receptor-deficient C57Bl/6J mice develop glomerulosclerosis . Young female mice on a C57Bl/6J ( B6 ) background are considered glomerulosclerosis ( GS ) -resistant but aging B6 mice develop mild GS . Estrogen deficiency accelerates while estrogen replacement retards GS in young sclerosis-prone oligosyndactyly mutant mice on an P04000 background . To explore the effects of sex hormones on glomerular structure and function in the context of gender and genetic background , we studied mice in which the estrogen-receptor ( ER ) genes alpha- or -beta were deleted ( alpha- or betaER knockout ( KO ) ) and crossed into the B6 background . We also studied ovariectomized ( Ovx ) B6 mice given testosterone . Male and female betaERKO and male alphaERKO mice had no glomerular dysfunction at 9 months of age ; however , alphaERKO female mice displayed albuminuria and GS . Ovx prevented glomerular dysfunction in alphaERKO female mice by eliminating endogenous testosterone production while exogenous testosterone induced GS in Ovx B6 mice . P10275 ( AR ) expression and function was found in microdissected glomeruli and cultured mesangial cells . DB00624 compared to placebo increased both AR expression and TGF-beta1 mRNA levels in glomeruli isolated from female B6 mice . Estrogen deficiency had no deleterious effects on the glomeruli in B6 mice . Our study shows that genetic traits strongly influence the GS-promoting effects of estrogen deficiency while testosterone induces GS in a gender-specific manner . P10275 in Sertoli cells is not required for testosterone-induced suppression of spermatogenesis , but contributes to Sertoli cell organization in Utp14b jsd mice . DB00624 acting through the androgen receptor ( AR ) maintains the arrest of spermatogonial differentiation in juvenile spermatogonial depletion ( jsd mutation in the Utp14b gene ) mutant adult male mice . It is not known which of the somatic cell types expressing AR mediates this inhibition . To determine whether Sertoli cells are responsible , we selectively eliminated AR in Sertoli cells in jsd mice containing a floxed-Ar gene and an anti-Müllerian hormone-Cre transgene . In these Sertoli AR-knockout ( SCARKO ) -jsd mice , spermatogonial differentiation did not recover . However , the normal organization of Sertoli cell nuclei was drastically disrupted in SCARKO-jsd mice compared with SCARKO or jsd mice . In addition , the extent of ectoplasmic specializations was reduced ; tight junctions were not found ; vinculin , an anchoring protein found in ectoplasmic specializations , became uniformly distributed in the cytoplasm ; and the adult Sertoli cells showed excess heterochromatin subjacent to their nuclear envelope . Despite the abnormalities in Sertoli cells in SCARKO-jsd mice , global suppression of testosterone action and levels was still effective in restoring the differentiated germ cells , and this was accompanied by an improved arrangement of Sertoli cell nuclei . We conclude that Sertoli cells are not targets for the testosterone-mediated inhibition of spermatogonial differentiation in jsd mice , and that both AR in Sertoli cells and the presence of differentiated germ cells contribute to maintaining the organization of Sertoli cells within the seminiferous tubules . Sex steroid regulation of chin-marking behavior in male New Zealand rabbits . Chin-marking behavior ( chinning ) was evaluated daily in nine intact adult male rabbits . All subjects ( Ss ) displayed chinning ( mean of means +/- SE = 61 +/- 7 marks/10 min ) but the frequency of this behavior varied largely across them ( range of mean chinning frequency = 19-84 marks/10 min ) . Chinning frequency showed abrupt variations at intervals of 2-3 days , but periodogram analysis did not reveal the existence of an endogenous rhythm in this behavior . Castration significantly decreased ( mean of means +/- SE = 29 +/- 9 marks/10 min ; p < 0.01 ) . but did not suppress chinning . DB00624 propionate ( TP ; 1 mg/day for 16 days ) restored chinning in castrated Ss to slightly below precastration levels ( mean +/- S.E. V 53 +/- 13 marks/10 min ) . The daily administration of 1 microgram estradiol benzoate ( EB ) plus 1 mg dihydrotestosterone propionate ( DHTP ) stimulated chinning within 2 days ( mean increase = 147 % ; p < 0.005 ) . DHTP ( 1 mg/day ) given alone stimulated chinning only after 11 days of treatment ( mean increase = 475 % ; p < 0.01 ) . At higher doses , both DHTP ( 10 mg/day ) and EB ( 10 or 50 micrograms/day ) stimulated chinning by 450 % , 80 % , and 100 % , respectively , over baseline values . Results indicate that chinning largely depends on testicular steroids . P10275 occupation by T or DB02901 , which is enhanced by E , optimally activates chinning . Increased dihydrotestosterone receptor levels in high-stage renal adenocarcinoma . Primary renal adenocarcinoma tissue , metastatic deposits , and normal kidney parenchyma from 16 patients were assayed for sex hormone receptors by dextran-coated charcoal adsorption and sucrose gradient centrifugation techniques . DB02901 receptors ( P10275 ) were found in all renal carcinomatous tissue ( 20/20 ) and in 93 % ( 13/14 ) autologous normal kidneys analyzed . DB00624 receptors were found in 84 % ( 16/19 ) of tumors and 93 % ( 14/15 ) or normal kidneys analyzed . Estrogen receptors in small amounts ( ER ) were detected in only 5 % ( 1/19 ) of tumors and in 7 % ( 1/15 ) of normal kidneys . Progesterone receptors ( PR ) in low quantities were detected in 30 % ( 6/20 ) of renal tumors and in 40 % ( 6/15 ) of normal kidneys . P10275 levels in high-stage tumors ( DB00279 , DB00451 ) were significantly elevated over levels in autologous normal kidney , whereas in low-stage tumors localized to the kidney ( T1 and P24752 tumors ) P10275 levels were not significantly different from autologous normal kidney . The mean levels of P10275 in high-stage kidney tumors were significantly elevated over levels in low-stage tumors ( P less than 0.001 ) . P10275 estimation in renal neoplasms may help in biologic staging of renal adenocarcinoma and could define a group of patients in whom anti-androgen therapy may be worth a trial . Altered levels of angiopoietin 1 and tie 2 are associated with androgen-regulated vascular regression and growth in the ventral prostate in adult mice and rats . The involution of the rat ventral prostate gland after castration could be caused by primary changes in the vasculature . To explore the mechanisms , we studied the effects of castration and testosterone treatment on the vasculature in the ventral prostate in adult rats and mice . P10275 expression , vascular morphology , and the expression of angiopoietin ( ang ) 1 and 2 and their receptor tie 2 were examined 1 , 3 , and 7 d after castration and after testosterone treatment of castrated animals using stereological methods , immunohistochemistry , laser capture microdissection , and Western blotting . One day after castration , the percentage of blood vessels covered with smooth muscle actin , endothelial cell proliferation , and vascular volume had decreased , whereas endothelial cell apoptosis had increased . Simultaneously , ang 1 and tie 2 protein levels decreased . Nuclear expression of androgen receptor was observed not only in glandular and stroma smooth muscle cells but also in the mural cells of prostate arteries and veins and was markedly down-regulated already 1 d after castration . DB00624 administration of castrated mice and rats reversed all the observed effects . At the mRNA level , tie 2 was exclusively , but ang 1 predominantly , expressed in the stroma , compared with the epithelial compartment . Local delivery of soluble tie 2 during testosterone-stimulated growth , inhibited vascular maturation and increased vascular volume and leukocyte infiltration compared with controls . We conclude that androgens may regulate the prostate vasculature by direct effects on mural vascular cells and by influencing the secretion of the angiopoietins , in above all , the stroma cells . Androgen receptors , serum androgen levels and survival of breast cancer patients . Steroid receptor levels and serum androgen levels were determined in 61 breast cancer patients and 34 patients with non-malignant breast lesions . DB00624 and dehydroepiandrosterone-sulfate did not and androstenedione did show a difference between the two groups . Androgen levels had no influence on survival rates . P10275 ( AR ) levels correlated with progesterone receptor levels , but not with estrogen receptor levels or with tumor stage . Patients with positive AR findings had a better survival rate ; this was independent of tumor stage . AR findings may therefore be a prognostic index in breast cancer patients . Differential role of Q07869 gamma in the regulation of P25874 -1 and adipogenesis by P01375 in brown adipocytes . Extracellular regulated kinases ( ERKs ) mediate the inhibitory effect of tumor necrosis factor alpha ( P01375 ) on uncoupling protein-1 ( P25874 -1 ) , but not on lipid accumulation . P01375 -induced P29323 -dependent peroxisome proliferator activator receptor gamma ( Q07869 gamma ) phosphorylation could be responsible for P25874 -1 downregulation . Thus , the negative effect of P01375 on P25874 -1 mRNA expression at 4-5 h , under basal conditions or in cells treated with the Q07869 gamma agonist , rosiglitazone , was reversed by the Q02750 inhibitor PD98059 . In contrast , fatty acid synthase and malic enzyme mRNA downregulation was not prevented . Moreover , rosiglitazone has no positive effect on adipogenic gene expression or lipid accumulation . Therefore , there is a differential regulation of thermogenic and adipogenic differentiation by Q07869 gamma , which might account for the differences in the P01375 regulation through ERKs . P10275 decreases CMYC and P01116 expression by upregulating let-7a expression in ER- , PR- , AR+ breast cancer . It is generally known that the decision to use anti-estrogen therapy is based on the expression of estrogen and progesterone receptors in breast cancers . Recent studies have shown that androgen receptor ( AR ) is frequently expressed in ER- , PR- breast cancer and plays an important role in the prognosis of breast cancer patients . Furthermore , AR can increase the global expression of microRNAs , post-transcriptional gene regulators that play a crucial role in the initiation and progression of breast cancer . In this study , we investigated the functions and relations of AR , related miRNAs and target proteins in ER- , PR- , AR+ breast cancer . The results showed that androgen-induced AR activating signal directly upregulates let-7a expression , downregulates CMYC and P01116 protein expression , and inhibits cell proliferation in ER- , PR- , AR+ breast cancer cells . Overexpression of let-7a inhibits cell proliferation and downregulates CMYC and P01116 protein expression , whereas inhibition of let-7a expression by specific antisense oligonucleo-tides increases cell growth and upregulates CMYC and P01116 protein expression . We performed in situ hybridization for let-7a and immunohistochemical staining for CMYC and P01116 using sequential sections obtained from surgically-resected breast cancer tissues and observed an inverse correlation between the staining pattern of let-7a and its target proteins . Androgen-induced AR activating signal upregulates let-7a that targets CMYC and P01116 and contributes to ER- , PR- , AR+ breast cancer pathogenesis . Elucidation of this pathway will help develop new therapies . P10275 signalling in peritubular myoid cells is essential for normal differentiation and function of adult Leydig cells . DB00624 synthesis depends on normal Leydig cell ( LC ) development , but the mechanisms controlling this development remain unclear . We recently demonstrated that androgen receptor ( AR ) ablation from a proportion of testicular peritubular myoid cells ( PTM-ARKO ) did not affect LC number , but resulted in compensated LC failure . The current study extends these investigations , demonstrating that PTM AR signalling is important for normal development , ultrastructure and function of adult LCs . Notably , mRNAs for LC markers [ e.g. steroidogenic factor 1 ( Nr5a1 ) , insulin-like growth factor ( Igf-1 ) and insulin-like factor 3 ( Insl3 ) ] were significantly reduced in adult PTM-ARKOs , but not all LCs were similarly affected . Two LC sub-populations were identified , one apparently ' normal ' sub-population that expressed adult LC markers and steroidogenic enzymes as in controls , and another ' abnormal ' sub-population that had arrested development and only weakly expressed P51460 , luteinizing hormone receptor , and several steroidogenic enzymes . Furthermore , unlike ' normal ' LCs in PTM-ARKOs , the ' abnormal ' LCs did not involute as expected in response to exogenous testosterone . Differential function of these LC sub-populations is likely to mean that the ' normal ' LCs work harder to compensate for the ' abnormal ' LCs to maintain normal serum testosterone . These findings reveal new paracrine mechanisms underlying adult LC development , which can be further investigated using PTM-ARKOs . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Hormonal influences of early postnatal indomethacin and ibuprofen in neonatal rats . OBJECTIVE : Indomethacin and ibuprofen are administered to preterm neonates for symptomatic patent ductus arteriosus . The drugs suppress prostaglandins ( PGs ) which modulate growth and secretion of various hormones . We examined the hypothesis that early postnatal indomethacin and ibuprofen influence growth and GH- P05019 -insulin and Q9Y251 axes in neonatal rats . DESIGN : Rat pups received IP injections of saline ( Sal ) on P1 , P2 , and P09131 ; 10mg/kg ibuprofen on P1 followed by 5mg/kg on P2 and P09131 ; or 0.2mg/kg indomethacin on P1 followed by 0.1mg/kg on P2 and P09131 . Serum and hepatic GH , P30043 and P05019 ; and serum corticosterone and insulin levels were determined . RESULTS : Ibuprofen suppressed somatic growth in the sucking rats , but the effect was transient , resolving by P14 . Indomethacin had an opposite , latent effect on body weight and liver to body weight ratios in weanling rats . Both indomethacin and ibuprofen had profound hormonal effects that differed in magnitude and timing . Indomethacin resulted in a sustained elevation in corticosterone levels at P21 , while ibuprofen increased serum and hepatic GH levels . Both drugs suppressed P30043 in serum at Q0GE19 and P14 ; and liver at P4 and Q0GE19 , but a rebound increase in serum P30043 was noted at P21 with Ibuprofen only . Both drugs increased serum P05019 at Q0GE19 . The effect remained sustained with indomethacin . CONCLUSIONS : These results provide evidence for an involvement of PGs in the regulation of growth as well as the GH-IGF and Q9Y251 axes . Therefore , early postnatal exposure to PG inhibitors may further exacerbate postnatal growth restriction and ability to cope with stress . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . Microarray analyses of the effects of NF-kappaB or PI3K pathway inhibitors on the LPS-induced gene expression profile in RAW264.7 cells : synergistic effects of rapamycin on LPS-induced P14780 -overexpression . Lipopolysaccharide ( LPS ) activates a broad range of signalling pathways including mainly NF-kappaB and the MAPK cascade , but recent evidence suggests that LPS stimulation also activates the PI3K pathway . To unravel the specific roles of both pathways in LPS signalling and gene expression profiling , we investigated the effects of different inhibitors of NF-kappaB ( BAY 11-7082 ) , PI3K ( wortmannin and LY294002 ) but also of P42345 ( rapamycin ) , a kinase acting downstream of PI3K/Akt , in LPS-stimulated RAW264.7 macrophages , analyzing their effects on the LPS-induced gene expression profile using a low density DNA microarray designed to monitor the expression of pro-inflammatory genes . After statistical and hierarchical cluster analyses , we determined five clusters of genes differentially affected by the four inhibitors used . In the fifth cluster corresponding to genes upregulated by LPS and mainly affected by BAY 11-7082 , the gene encoding P14780 displayed a particular expression profile , since rapamycin drastically enhanced the LPS-induced upregulation at both the mRNA and protein levels . DB00877 also enhanced the LPS-induced NF-kappaB transactivation as determined by a reporter assay , phosphorylation of the p38 and Erk1/2 MAPKs , and counteracted Q07869 activity . These results suggest that P42345 could negatively regulate the effects of LPS on the NF-kappaB and MAPK pathways . We also performed real-time RT-PCR assays on mmp9 expression using rosiglitazone ( agonist of PPARgamma ) , PD98059 ( inhibitor of Erk 1/2 ) and SB203580 ( inhibitor of p38(MAPK) ) , that were able to counteract the rapamycin mediated overexpression of mmp9 in response to LPS . Our results suggest a new pathway involving P42345 for regulating specifically mmp9 in LPS-stimulated RAW264.7 cells . DB02901 interacts with P00533 /MAPK signalling and modulates P00533 levels in androgen receptor-positive LNCaP prostate cancer cells . P10275 ( AR ) signalling plays a pivotal role in prostate cancer pathogenesis and progression . However , androgen-mediated AR signalling is yet to be fully understood . P00533 and Q96HU1 kinase signalling pathways play predominant roles in AR function . Therefore , we investigated the interaction of P00533 signalling and AR activity in AR-positive LNCaP cells . We found that 5alpha-dihydrotestosterone ( DB02901 ) and P01133 had a synergistic effect on AR activity as detected by a luciferase reporter system , although P01133 alone did not activate AR . Both P27361 /2 and p38 were involved in DB02901 and DB02901 / P01133 -induced AR activation as detected by specific MEK and p38 inhibitors . Furthermore , 24-h treatment of the cells with DB02901 resulted in ubiquitination and down-regulation of the P00533 . This effect could be inhibited by the anti-androgen flutamide , suggesting an androgen-dependent mechanism . On the other hand , DB02901 -treatment strongly increased AR levels in LNCaP cells . These data suggest a complex regulatory loop between activated AR and P00533 . In conclusion , activation of AR by both DB02901 and P01133 / DB02901 involves the Q96HU1 kinase pathway . Long-term activation of AR results in increase of AR levels , which through so far unknown regulatory mechanisms results in ubiquitination and degradation of the P00533 . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . P10275 is essential for sexual differentiation of responses to olfactory cues in mice . During sexual differentiation males and females are exposed to different levels of testosterone , which promotes sex differences in the adult brain and in behavior . DB00624 can act after aromatization or reduction via a number of steroid hormone receptors . Here we provide new evidence that the androgen receptor ( AR ) is essential for sexual differentiation in mice . We used mice carrying the testicular feminization ( Tfm ) mutation of the AR . Adult Tfm males , wild-type male and female littermates were gonadectomized and given subcutaneous estradiol implants . In all sexually dimorphic traits , Tfm males had responses equivalent to females and different from males . In simultaneous choice tests , males spent significantly more time investigating female-soiled bedding , whereas females and Tfm males preferred to investigate male-soiled bedding . Tfm males and females did not have a partner preference in tests with awake stimulus animals , whereas males showed a preference for females over males . Exposure to male-soiled , but not clean , bedding produced a significant increase in c-Fos-immunoreactive cells in the medial preoptic area and bed nucleus of the stria terminalis in Tfm males and females , no increase was noted in males . Masculine sexual behavior ( mounting and thrusting ) was not sexually dimorphic , and all groups displayed these behaviors . Our results support data collected in humans suggesting a role for the androgen receptor in sexual differentiation of social preferences and neural responses to pheromones . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . P10275 up-regulates insulin-like growth factor binding protein-5 ( P24593 ) expression in a human prostate cancer xenograft . The insulin-like growth factor ( IGF ) binding proteins ( IGFBPs ) are important modulators of IGF action in many tissues including human prostate . IGFBPs and the androgen receptor ( AR ) are expressed in CWR22 , an androgen-dependent epithelial cell human CaP xenograft that retains biological characteristics of human CaPs , including regression following androgen withdrawal and recurrent growth of AR-containing cells in the absence of testicular androgens beginning several months after castration . Northern blot and in situ hybridization analyses demonstrated that P24593 is androgen-regulated in CWR22 . P24593 messenger RNA ( mRNA ) decreased by 90 % following castration of tumor-bearing mice compared with noncastrate androgen-stimulated mice . DB00624 treatment of CWR22 tumor-bearing mice 6 or 12 days after castration increased P24593 mRNA 10- to 12-fold . Levels of other IGFBP mRNAs did not change following androgen withdrawal and replacement . P24593 protein in tumor extracts bound 125I-labeled P05019 in ligand blot assays and the amounts of P24593 measured by immunoblotting paralleled the levels of P24593 mRNA . Androgen-induced expression of P24593 was at a maximum level within 24 h after testosterone replacement , whereas the major increase in cell proliferation as measured by Ki-67 immunostaining occurred between 24-48 h . This time course suggested P24593 may be a mediator of androgen-induced growth of CWR22 . In tumors that recurred several months following castration , P24593 mRNA and protein increased to levels that approached those in androgen-stimulated CWR22 tumors from noncastrate mice . P24593 immunohistochemical staining of prostate tissue specimens from patients was stronger in androgen-dependent and androgen-independent CaP than in areas of intraepithelial neoplasia ( P63167 ) or benign prostatic hyperplasia ( BPH ) . P24593 mRNA in these specimens was localized predominantly to stromal cells and P24593 protein to epithelial cell membranes . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men . Axotomy transiently down-regulates androgen receptors in motoneurons of the spinal nucleus of the bulbocavernosus . DB00624 is an important trophic factor for motoneurons in the spinal nucleus of the bulbocavernosus ( SNB ) , and SNB motoneurons are more responsive to testosterone than are other motoneurons . Axonal injury during early postnatal life prevents the normal development of steroid-sensitivity by adult SNB motoneurons . Axonal injury also causes changes in the expression by motoneurons of a wide range of proteins , including the up-regulation of trophic factor receptors . We have used a polyclonal antibody ( PG-21 ; G.S. Prins ) to study the expression of androgen receptors in SNB motoneurons after axonal injury . PG-21 labeled motoneuronal nuclei in the lower lumbar spinal cord of rats in a pattern that matched autoradiographic reports of androgen accumulation in this region of the nervous system . A population of numerous , small cells located dorsal to the central canal also showed evidence of androgen receptor expression . Cutting the axons of SNB motoneurons in adulthood or in development caused a decrease in androgen receptor immunoreactivity in SNB motoneurons . This is the first report that a trophic factor receptor in motoneurons is down-regulated after axonal injury , and is interesting in light of reports that testosterone treatment can facilitate motoneuronal regeneration after nerve cut . P10275 levels subsequently returned to normal , regardless of the age at axotomy , providing no evidence for a lasting effect of developmental axotomy on androgen receptor levels in SNB motoneurons . Thus , axotomy-induced down-regulation of androgen receptors does not underlie the inability of SNB motoneurons to respond to androgen treatment several months after pudendal nerve cut in development . P10275 dependent and independent activities of testosterone on hepatic microsomal drug metabolism . Administration of testosterone for 6 days to intact female and castrate male BALB/cJ mice stimulated hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content by 50-75 % . DB00624 also stimulated hepatic microsomal NADPH-oxidase activity , but to a lesser degree . To probe the mechanism of this effect of androgens , two antiandrogens ( cyproterone acetate and flutamide ) were employed . Since cyproterone acetate was a potent stimulator of hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content , no antiandrogenic activity of this steroid could be detected . By contrast , flutamide alone had little effect on either ethylmorphine N-demethylase activity or cytochrome P-450 content . However , this drug effectively blocked the stimulatory effects of testosterone on ethylmorphine N-demethylase activity and cytochrome P-450 content but not on NADPH-oxidase activity . This effect was not species specific , since flutamide also prevented androgen stimulation of ethylmorphine metabolism in adult castrate and prepubertal male Fisher rats . The testosterone-induced increase of hepatic weight and microsomal protein content was not affected by the administration of flutamide . The observations are consistent with the hypothesis that androgens have two distinct effects on the liver . First , testosterone may act as a general , nonspecific stimulant of liver weight and microsomal protein content which is independent of the androgen receptor . Secondly , testosterone action in the liver may be expressed via an androgen-specific or androgen receptor-dependent mechanism which controls , in part , the cytochrome P-450-dependent demethylase system . Pubertal maturation is associated with an increase in the number of androgen receptor-immunoreactive cells in the brains of male ferrets . P10275 -immunoreactive ( AR-IR ) cells were identified in brains of male ferrets before and after the onset of pubertal maturation . There was a greater number of AR-IR cells after the onset of pubertal maturation in some , but not all , brain regions examined . Regions in which the number of AR-IR cells increased included the preoptic area and the amygdala , areas known to be involved in the control of male reproductive behaviors . The mechanisms responsible for the increase in AR-IR cells are unknown , but might be related to the higher circulating levels of testosterone that were present in the older animals . DB00624 may increase androgen receptor ( AR ) immunoreactivity both by concentrating already existing ARs within the nucleus and by stimulating de novo synthesis of receptor protein . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . Ursolic acid inhibits the initiation , progression of prostate cancer and prolongs the survival of TRAMP mice by modulating pro-inflammatory pathways . Prostate cancer is one of the leading causes of cancer death among men worldwide . In this study , using transgenic adenocarcinoma of mouse prostate ( TRAMP ) mice , the effect of diet enriched with 1 % w/w ursolic acid ( UA ) was investigated to evaluate the stage specific chemopreventive activity against prostate cancer . We found that TRAMP mice fed with UA diet for 8 weeks ( weeks 4 to 12 ) delayed formation of prostate intraepithelial neoplasia ( P63167 ) . Similarly , mice fed with UA diet for 6 weeks ( weeks 12 to 18 ) inhibited progression of P63167 to adenocarcinoma as determined by hematoxylin and eosin staining . Finally , TRAMP mice fed with UA diet for 12 weeks ( weeks 24 to 36 ) demonstrated markedly reduced tumor growth without any significant effects on total body weight and prolonged overall survival . With respect to the molecular mechanism , we found that UA down-regulated activation of various pro-inflammatory mediators including , NF-κB , P40763 , AKT and IKKα/β phosphorylation in the dorsolateral prostate ( Q9NX01 ) tissues that correlated with the reduction in serum levels of P01375 -α and P05231 . In addition , UA significantly down-regulated the expression levels of cyclin D1 and P35354 but up-regulated the levels of caspase-3 as revealed by immunohistochemical analysis of tumor tissue sections . Finally , UA was detected in serum samples obtained from various mice groups fed with enriched diet in nanogram quantity indicating that it is well absorbed in the GI tract . Overall , our findings provide strong evidence that UA can be an excellent agent for both the prevention and treatment of prostate cancer . P10275 CAG repeat polymorphism is associated with serum testosterone levels , obesity and serum leptin in men with type 2 diabetes . OBJECTIVE : To determine the relationships between androgen receptor CAG repeat polymorphism length ( AR CAG ) , sex hormones and clinical variables in men with type 2 diabetes ( DM2 ) . Men with DM2 are known to have a high prevalence of low testosterone levels . Studies suggest that testosterone replacement therapy may improve insulin sensitivity and glycaemic control in men with DM2 and reduces central obesity and serum leptin . AR CAG is known to correlate negatively with AR sensitivity and positively with body fat , insulin levels , and leptin in healthy men . DESIGN : Cross-sectional study set in a district general hospital diabetes centre . METHODS : Sex hormones , AR CAG and symptoms of hypogonadism were assessed in 233 men with DM2 . Associations were sought between these variables and others such as obesity , leptin , glycaemic control , and blood pressure . RESULTS : DB00624 was negatively associated and AR CAG positively associated with obesity and leptin . The associations of AR CAG with leptin and obesity were independent of testosterone , estradiol , gonadotropins , and age . AR CAG was also independently associated with total , bioavailable and free testosterone , LH , waist circumference , body mass index , leptin , and systolic blood pressure . There was no association of AR CAG with sex hormone binding globulin , estradiol , HbA(1C) or the symptoms of hypogonadism . CONCLUSIONS : The association of longer AR CAG with obesity and leptin suggests that shorter AR CAG may have an influence in maintaining healthy anthropomorphics and metabolism in men with DM2 . DB00624 and LH levels are higher in men with longer AR CAG , probably reflecting reduced negative feedback through a less sensitive receptor . DB00624 stimulates proliferation and inhibits interleukin-6 production of normal and hereditary gingival fibromatosis fibroblasts . Hereditary gingival fibromatosis ( P14210 ) is a rare oral condition characterized by a slow and progressive enlargement of the gingiva , involving both the maxilla and mandible . In vitro , P14210 fibroblasts demonstrate a proliferative index significantly higher than fibroblasts from normal gingiva ( NG ) . The objective of this study was to determine the effect of dihydrotestosterone on the proliferation of gingival fibroblasts derived from patients with P14210 ( n = 4 ) and from four healthy individuals . Additionally , we analyzed the effect of dihydrotestosterone on interleukin-6 ( P05231 ) production and determined the expression levels of androgen receptors in NG and P14210 fibroblasts . Gingival fibroblasts from NG and P14210 were incubated with increasing concentrations of dihydrotestosterone with or without androgen blockers , and cultured for 24 h , and the proliferation index was determined by automated cell counter . P05231 production , in this system , was quantified using a " capture " enzyme-linked immunosorbent assay ( ELISA ) . Semi-quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR ) was performed to measure the mRNA expression of androgen receptors . The results indicated that dihydrotestosterone simultaneously downregulates the production of P05231 and upregulates the cell proliferation . DB01216 and cyprosterone acetate , two anti-androgens , partially reversed these effects . P10275 mRNA expression was identified in both NG and P14210 fibroblasts ; however , the levels in NG were higher than those observed in P14210 . These results show that testosterone coordinates the proliferation and production of P05231 of normal and P14210 fibroblasts . Androgen Receptor Enhances p27 Degradation in Prostate Cancer Cells through Rapid and Selective Q53ET0 Activation . P10275 ( AR ) plays a central role in prostate cancer ( PCa ) growth , with androgen deprivation or AR down-regulation causing cell-cycle arrest and accumulation of the p27 cyclin-dependent kinase inhibitor . The molecular basis for this AR regulation of cell-cycle progression remains unclear . Here we demonstrate that androgen can rapidly reduce p27 protein in PCa cells by increasing its proteasome-mediated degradation . This rapid androgen-stimulated p27 degradation was mediated by AKT through the phosphorylation of p27 T157 . Significantly , androgen increased Q53ET0 -mediated AKT S473 phosphorylation without affecting the PDK1-mediated AKT T308 phosphorylation or Q6UUV9 activity . The Q53ET0 activation was further supported by enhanced P42345 / Q6R327 association and increased phosphorylation of additional Q53ET0 substrates , O00141 and PKCα . The androgen-stimulated nuclear translocation of AR was associated with markedly-increased nuclear Q9BPZ7 , a critical component of Q53ET0 . Finally , the androgen-mediated Q53ET0 /AKT activation targets a subset of AKT substrates including p27 and Q12778 , but not Q96B36 . This study reveals a pathway linking AR to a selective activation of Q53ET0 , the subsequent activation of AKT , and phosphorylation of a discrete set of AKT substrates that regulate cellular proliferation and survival . These findings establish that Q53ET0 can function as a central regulator of growth in response to signals that are distinct from those regulating Q6UUV9 , and support efforts to target Q53ET0 for cancer therapy . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . H2 relaxin is a biased ligand relative to H3 relaxin at the relaxin family peptide receptor 3 ( Q9NSD7 ) . Relaxin family peptide 3 receptors ( Q9NSD7 ) are activated by H3-relaxin to inhibit forskolin-stimulated DB02527 accumulation and stimulate extracellular signal-regulated kinase ( P29323 ) 1/2 phosphorylation . In this study , we sought to identify novel signaling pathways coupled to Q9NSD7 and to investigate whether other members of the relaxin peptide family activated these pathways . Two patterns of signaling were observed in Q9NSD7 -expressing Chinese hamster ovary ( CHO ) - P04264 and human embryonic kidney ( P29320 ) -293 cells ( CHO- Q9NSD7 and P29320 - Q9NSD7 ) and murine septal neuron SN56 cell lines : 1 ) strong inhibition of forskolin-stimulated DB02527 accumulation , P27361 /2 activation and nuclear factor ( NF ) -kappaB reporter gene activation in cells stimulated with H3 relaxin , with weaker activity observed for H2 relaxin , porcine relaxin , or insulin-like peptide ( INSL ) 3 and 2 ) strong stimulation of activator protein ( AP ) -1 reporter genes by H2 relaxin , with weaker activation observed with H3 or porcine relaxin . Two distinct ligand binding sites were identified on Q9NSD7 -expressing cells using two different radioligands . (125)I- Q9Y5Q6 A-chain/relaxin-3 B-chain chimera bound with high affinity to the Q9NSD7 -expressing cells with competition by H3 relaxin or a H3 relaxin B-chain dimeric peptide , consistent with previous reports . Binding studies with (125)I-H2 relaxin revealed a distinct binding site with potent competition observed with H2 relaxin , H3 relaxin , or P51460 and weaker competition with porcine relaxin . Thus H3 relaxin potently activates all signaling pathways coupled to Q9NSD7 , whereas H2 relaxin is an AP-1-biased ligand relative to H3 relaxin . Androgens and metabolic syndrome : lessons from androgen receptor knock out ( ARKO ) mice . DB00624 ( T ) is an important factor for determining body composition in males . Abdominal obesity is inversely correlated with serum T levels in men , leading to greater mortality . Pathologically hypogonadal men also have a significantly higher fat mass , which is reversed by T administration . However , the mechanism for such anti-obesity effect of androgen has not been well clarified . P10275 ( AR ) null male mice revealed late-onset obesity . Male ARKO mice were euphagic compared to the wild-type male controls , but also less dynamic and less oxygen consuming . Transcript profiling indicated that male ARKO mice had lower transcripts for the thermogenetic uncoupling protein 1 ( P25874 ) . We also found enhanced secretion of adiponectin , which is insulin-sensitizing , from adipose tissue in comparison to wild type , which might partly explain why the overall insulin sensitivity of male ARKO mice remained almost intact despite their apparent obesity . In addition , decreased lipolysis rather than increased lipid synthesis was observed , which might also account for the increased adiposity in male ARKO mice . The results revealed that AR plays important roles in male metabolism by affecting the energy balance , and is negative to both adiposity and insulin sensitivity . DB00624 potentiates the hypoxic ventilatory response of adult male rats subjected to neonatal stress . Neonatal stress disrupts development of homeostatic systems . During adulthood , male rats subjected to neonatal maternal separation ( Q5H8A3 ) are hypertensive and show a larger hypoxic ventilatory response ( HVR ) , with greater respiratory instability during sleep . Neonatal stress also affects sex hormone secretion ; hypoxia increases circulating testosterone of Q5H8A3 ( but not control ) male rats . Given that these effects of Q5H8A3 are not observed in females , we tested the hypothesis that testosterone elevation is necessary for the stress-related increase of the HVR in adult male rats . Pups subjected to Q5H8A3 were placed in an incubator for 3 h per day from postnatal day 3 to 12 . Control pups remained undisturbed . Rats were reared until adulthood , and the HVR was measured by plethysmography ( fractional inspired O2 = 0.12 , for 20 min ) . We used gonadectomy to evaluate the effects of reducing testosterone on the HVR . Gonadectomy had no effect on the HVR of control animals but reduced that of Q5H8A3 animals below control levels . Immunohistochemistry was used to quantify androgen receptors in brainstem areas involved in the HVR . P10275 expression was generally greater in Q5H8A3 rats than in control rats ; the most significant increase was noted in the caudal region of the nucleus tractus solitarii . We conclude that the abnormal regulation of testosterone is important in stress-related augmentation of the HVR . The greater number of androgen receptors within the brainstem may explain why Q5H8A3 rats are more sensitive to testosterone withdrawal . Based on the similarities of the cardiorespiratory phenotype of Q5H8A3 rats and patients suffering from sleep-disordered breathing , these results provide new insight into its pathophysiology , especially sex-based differences in its prevalence . Mutation of the calmodulin binding motif IQ of the L-type Ca(v)1.2 Ca2+ channel to EQ induces dilated cardiomyopathy and death . Cardiac excitation-contraction coupling ( EC coupling ) links the electrical excitation of the cell membrane to the mechanical contractile machinery of the heart . DB01373 channels are major players of EC coupling and are regulated by voltage and Ca(2+)/calmodulin ( P62158 ) . P62158 binds to the IQ motif located in the C terminus of the Ca(v)1.2 channel and induces Ca(2+)-dependent inactivation ( CDI ) and facilitation ( P05231 ) . Mutation of DB00167 to DB00142 ( Ile1624Glu ) in the IQ motif abolished regulation of the channel by CDI and P05231 . Here , we addressed the physiological consequences of such a mutation in the heart . Murine hearts expressing the Ca(v)1.2(I1624E) mutation were generated in adult heterozygous mice through inactivation of the floxed WT Ca(v)1.2( Q401N2 ) allele by tamoxifen-induced cardiac-specific activation of the MerCreMer Cre recombinase . Within 10 days after the first tamoxifen injection these mice developed dilated cardiomyopathy ( DCM ) accompanied by apoptosis of cardiac myocytes ( CM ) and fibrosis . In Ca(v)1.2(I1624E) hearts , the activity of phospho- P62158 kinase II and phospho-MAPK was increased . CMs expressed reduced levels of Ca(v)1.2(I1624E) channel protein and I(Ca) . The Ca(v)1.2(I1624E) channel showed " CDI " kinetics . Despite a lower sarcoplasmic reticulum Ca(2+) content , cellular contractility and global Ca(2+) transients remained unchanged because the EC coupling gain was up-regulated by an increased neuroendocrine activity . Treatment of mice with metoprolol and captopril reduced DCM in Ca(v)1.2(I1624E) hearts at day 10 . We conclude that mutation of the IQ motif to IE leads to dilated cardiomyopathy and death . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . P42345 inhibition as therapy for hematologic malignancies . The mammalian target of rapamycin ( P42345 ) is a downstream effector of the phosphatidylinositol 3-kinase ( PI3K ) /Akt ( protein kinase B ) signaling pathway , which mediates cell survival and proliferation . P42345 regulates essential signal-transduction pathways , is involved in the coupling of growth stimuli with cell cycle progression , and initiates mRNA translation in response to favorable nutrient environments . P42345 is involved in regulating many aspects of cell growth , including membrane traffic , protein degradation , protein kinase C signaling , ribosome biogenesis , and transcription . Because P42345 activates both the P62753 kinase ( p70s6k ) and the eukaryotic initiation factor 4E-binding protein 1 , its inhibitors cause P55008 -phase cell cycle arrest . Inhibitors of P42345 also prevent cyclin dependent kinase ( CDK ) activation , inhibit retinoblastoma protein phosphorylation , and accelerate the turnover of cyclin D1 , leading to a deficiency of active P11802 /cyclin D1 complexes , all of which may help cause P55008 -phase arrest . It is known that the phosphatase and tensin homologue tumor suppressor gene ( P60484 ) plays a major role in embryonic development , cell migration , and apoptosis . Malignancies with P60484 mutations , which are associated with constitutive activation of the PI3K/Akt pathway , are relatively resistant to apoptosis and may be particularly sensitive to P42345 inhibitors . DB00877 analogs with relatively favorable pharmaceutical properties , including CCI-779 , RAD001 , and AP23573 , are under investigation in patients with hematologic malignancies . D- DB00128 implication in the modulation of frog brain sex steroid levels . There is evidence that D-aspartate ( D- DB00128 ) modulates sex hormone levels in frog testis by regulating the activity of P450 aromatase ( P450 aro ) , the key enzyme which converts DB00624 ( T ) in 17ß-Estradiol ( E2 ) . Here we report , for the first time , that there is a direct correlation among brain levels of D- DB00128 , P450 aro , E2 and Estradiol Receptor ( ERα ) in the male frogs during the reproductive as well as the post-reproductive phases of the breeding cycle , with highest levels being observed in the post-reproductive period . D- DB00128 i.p. administration to frogs ready for reproduction , induced an increase of brain P450 aro protein expression with concomitant enhancement of both E2 levels and ERα expression ; at the same time , brain T levels and P10275 expression decreased . In contrast , in the post-reproductive frogs , D- DB00128 treatment did not modify any of these parameters . Taken together , these results imply that the regulation of P450 aro expression by D- DB00128 could be an important step in the control of E2 levels in the frog brain . Serum testosterone plays an important role in the metastatic ability of castration resistant prostate cancer . PURPOSE : Prostate cells are dependent on androgens for growth and proliferation . Androgen deprivation therapy is the recommended treatment for advanced/metastatic prostate cancer . Under this therapy , prostate cancer will inevitably progress to castration resistant prostate cancer ( CRPC ) . Despite putative castration resistance , testosterone might still play a crucial role in the progression of CRPC . The goal of this study was to determine the role of testosterone in the formation of metastases of CRPC in both in vitro and in vivo settings . METHODS : In vitro , the effect of testosterone and the non-aromatizable androgen methyltrienolone on migration , invasion and proliferation of a castration-resistant prostate cancer rat cell line ( Dunning R3327-MATLyLu ) was assessed using a transwell assay and a sulforhodamine B assay and immunohistochemical detection of ki67 . P10275 status was determined using Western blot . In vivo , Copenhagen rats were divided in four groups ( males , females , castrated males and females with testosterone suppletion ) and inoculated with MATLyLu cells . Tumor size was assessed daily . RESULTS : DB00624 increased cell migration and invasion in a concentration-dependent manner in vitro . DB00624 did not affect in vitro cell proliferation . No difference was shown between the effect of testosterone and methyltrienolone . In vivo , in groups with higher levels of circulating testosterone , more rats had (micro)metastases compared with groups with low levels of testosterone . No effect was observed on primary tumor size/growth . CONCLUSIONS : Despite assumed castration resistance , progression of prostate cancer is still influenced by androgens . Therefore , continuous suppression of serum testosterone in patients who show disease progression during castration therapy is still warranted . P21554 regulates P29323 and GSK-3β-dependent glucocorticoid inhibition of osteoblast differentiation in murine MC3T3-E1 cells . Supraphysiological glucocorticoid administration accelerates loss of survival and differentiation in osteoblastic cells , thereby increasing the risks of osteopenic or osteonecrotic disorders . Neuroendocrine component type 1 cannabinoid receptor ( P21554 ) is found to regulate bone mass . This study characterized the biological role of P21554 in glucocorticoid-induced suppression of osteoblast differentiation . Murine MC3T3-E1 osteoblasts were incubated under osteogenic conditions in the presence or absence of 1 μM glucocorticoid , RNA interference , P21554 antagonist AM251 , and agonist WIN55212-2 . Cell survival was detected by formazan synthesis and TUNEL staining . Osteoblast differentiation was quantified by mineralized matrix accumulation and expression of the osteogenic factors Runx2 and osteocalcin . Expression of signaling molecules was assessed by immunoblotting . Glucocorticoid increased P21554 expression in association with decreased osteocalcin expression and mineralized nodule deposition . P21554 RNA interference and AM251 attenuated the deleterious actions of glucocorticoid treatment on survival and osteogenic activities , whereas activating P21554 by WIN55212-2 impaired osteoblast differentiation . P21554 signaling regulated JNK , P29323 , GSK-3β , and Akt activation as well as Runx2 and P05019 expression . Inhibition of GSK-3β by the kinase-inactive GSK-3β mutant or activation of P29323 by the active MEK-1 mutant abrogated glucocorticoid-induced inhibition of osteoblast differentiation . Glucocorticoid-induced P21554 expression occurred via glucocorticoid receptor-dependent transcriptional and translational regulation . Gain of Runx2 function and loss of P28562 action attenuated glucocorticoid-induced enhancement of P21554 expression . Taken together , P21554 regulation of P29323 and GSK-3β-dependent pathways participates in glucocorticoid inhibition of Runx2 signaling and osteoblast differentiation . Runx2 reciprocally regulates glucocorticoid-induced promotion of P21554 signaling . Our findings provide new insights into the role of the neuroendocrine component P21554 in glucocorticoid-induced osteoblast dysfunction .

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Design of novel potent antihyperlipidemic agents with antioxidant/anti-inflammatory properties : exploiting phenothiazine 's strong antioxidant activity . Because atherosclerosis is an inflammatory process involving a series of pathological events such as dyslipidemia , oxidative stress , and blood clotting mechanisms , we hereby report the synthesis and evaluation of novel compounds in which antioxidant , anti-inflammatory , and squalene synthase ( P37268 ) inhibitory/hypolipidemic activities are combined in simple molecules through design . The coupling of two different pharmacophores afforded compounds 1-12 , whose biological profile was markedly improved compared to those of parent lead structures ( i.e. , the hypolipidemic 2-hydroxy-2-aryl-(benzo)oxa ( or thia ) zine and the antioxidant phenothiazine ) . Most derivatives strongly inhibited in vitro microsomal lipid and LDL peroxidation , exhibiting potent free-radical scavenging activity . They further significantly inhibited P37268 activity and showed remarkable antidyslipidemic activity in vivo in animal models of acute and high-fat-induced hyperlipidemia . Finally , several compounds showed anti-inflammatory activity in vitro , inhibiting cycloxygenase ( P23219 /2 ) activity . The multimodal properties of the new compounds and especially their combined antioxidant/ P37268 / P36551 inhibitory activity render them interesting lead compounds for further evaluation against atherosclerosis . Evaluation of pharmacological profile of meloxicam as an anti-inflammatory agent , with particular reference to its relative selectivity for cyclooxygenase-2 over cyclooxygenase-1 . We studied the anti-inflammatory activity of meloxicam on rat carrageenin-induced pleurisy and its toxicity for rat gastric mucosa , relative to its in vitro inhibitory potency against partially purified cyclooxygenase ( P36551 ) -1 and P35354 preparations in order to clarify the pharmacological profile of the compound as an anti-inflammatory agent . In rat carrageenin-induced pleurisy , the plasma exudation rate peaked at 5 h , at which time P35354 was detectable in cells from the pleural exudate . Meloxicam and piroxicam ( 1 and 3 mg/kg ) and NS-398 ( 3 mg/kg ) showed almost equal anti-inflammatory potency against 5-hour pleurisy . A single oral administration of the compounds caused a dose-dependent increase in the number of rats with gastric mucosal erosion . The ED50 value for meloxicam ( 5.92 mg/kg ) was significantly higher than that for piroxicam ( 1.76 mg/kg ) , indicating that meloxicam is safer . DB00328 showed intermediate safety ( 2.59 mg/kg ) . In in vitro experiments , indometacin inhibited P23219 about 1.7 times more potently than P35354 . NS-398 inhibited P35354 with an IC50 of 0.32 microM , but never affected P23219 activity , even at 100 microM . In the same assay system , meloxicam inhibited P35354 about 12 times more selectively than P23219 . Piroxicam , however , inhibited both isoforms almost equally . These results indicate that meloxicam is a potent anti-inflammatory agent with low gastric toxicity . One reason for its in vivo pharmacological profile may be related to its relative selectivity for P35354 over P23219 . Thus , meloxicam may belong to a group of P35354 selective anti-inflammatory agents with a better safety profile than conventional P23219 and P35354 nonselective anti-inflammatory agents . Histone deacetylase inhibitors suppress the induction of c-Jun and its target genes including P35354 . P35354 ( P35354 ) is considered to be a target for anticancer therapy . Histone deacetylase ( HDAC ) inhibitors exhibit antitumor activity , but the mechanisms of action are incompletely understood . We investigated whether HDAC inhibitors blocked AP-1-mediated activation of P35354 transcription . Trichostatin A and suberoylanilide hydroxamic acid , two structurally related inhibitors of HDAC activity , blocked AP-1-mediated induction of P35354 expression and prostaglandin E2 biosynthesis . Chromatin immunoprecipitation assays indicated that HDAC inhibitors suppressed c-Jun binding to the P35354 promoter and thereby blocked transcription . The observed reduction in binding reflected reduced levels of c-Jun . HDAC inhibitors suppressed the induction of c-jun transcription by blocking the recruitment of the preinitiation complex ( RNA polymerase II and Q00403 ) to the c-jun promoter . O15379 but not Q13547 or Q92769 was required for AP-1-mediated stimulation of c-jun expression . Because HDAC inhibitors suppressed the induction of c-jun gene expression , resulting in reduced P35354 transcription , it was important to determine whether other known AP-1 target genes were also modulated . P12004 D1 and collagenase-1 are AP-1-dependent genes that have been implicated in carcinogenesis . HDAC inhibitors suppressed the induction of both cyclin D1 and collagenase-1 transcription by inhibiting the binding of c-Jun to the respective promoters . Taken together , these results suggest that HDAC inhibitors block the induction of c-jun transcription by inhibiting the recruitment of the preinitiation complex to the c-jun promoter . This led , in turn , to reduced expression of several activator protein-1-dependent genes ( P35354 , cyclin D1 , collagenase-1 ) . These findings provide new insights into the mechanisms underlying the antitumor activity of HDAC inhibitors . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . A 3-D model for P08908 -receptor agonists based on stereoselective methyl-substituted and conformationally restricted analogues of 8-hydroxy-2-(dipropylamino)tetralin . The enantiomers of cis- and trans-1,2,3,4,4a,5,10,10a-octahydro-9-hydroxy-1- propylbenzo[g]quinolines ( 10 and 11 , respectively ) and the enantiomers of trans-1,2,3,4,4a,5,6,10b-octahydro-10- hydroxy-4-propylbenzo[f]quinoline ( 12 ) have been synthesized and their stereochemical and conformational characteristics have been studied by use of X-ray crystallography and molecular mechanics ( P08253 ) calculations . The compounds , which are conformationally restricted analogues of the potent 5-hydroxytryptamine ( 5-HT ) receptor agonist 8-hydroxy-2- (dipropylamino)tetralin ( 8-OH-DPAT ; 1 ) have been evaluated for central 5-HT and dopamine receptor stimulating activity by use of biochemical and behavioral tests in rats . In addition , we have evaluated the ability of these compounds and a number of previously reported analogues to displace [ 3H ] -8-OH-DPAT from P08908 -binding sites . The enantiomers of 12 behave as potent P08908 -receptor agonists , whereas the octahydrobenzo[g]quinoline derivatives are much less potent or inactive . In general , the affinities of the compounds correlate well with their agonist potencies . The set of compounds under study is accommodated by a novel computer-graphics-derived model for P08908 -receptor agonism . The model consists of a flexible pharmacophore and a partial receptor-excluded volume . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . Transgenic Panax ginseng inhibits the production of P01375 , P05231 , and P10145 as well as P35354 expression in human mast cells . The most well-known medicinal plant , Panax ginseng ( P. ginseng ) , contains various phytosterols and bioactive triterpene saponins ( ginsenosides ) . P37268 is a key regulatory enzyme for triterpene biosynthesis and overexpression of the squalene synthase confers the hyper-production of triterpene saponins to form transgenic ginseng . In this study , we have investigated whether and how transgenic P. ginseng modulates an inflammatory reaction in a stimulated human mast cell line , HMC-1 . It was found that transgenic P. ginseng inhibited the production of tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -6 , P10145 , and the expression of cyclooxygenase-2 in phorbol 12-myristate 13-acetate ( PMA ) plus calcium ionophore A23187 ( PMACI ) -stimulated HMC-1 . Additionally , we have shown that transgenic P. ginseng suppressed the intracellular calcium level induced by PMACI . These results provide new insights into the pharmacological actions of transgenic P. ginseng as a potential molecule for use in therapy in mast cell-mediated inflammatory diseases . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Effects of cyclosporin and FK-506 on glomerular mesangial cells . Evidence for direct inhibition of thromboxane synthase by low cyclosporin concentrations . The cellular sources or molecular mechanisms responsible for the derangement of vasoactive prostanoid levels during immunosuppressive cyclosporin ( Q13216 ) therapy have not been defined . Using cultured rat glomerular mesangial cells ( MC ) , the cytostatic , cytotoxic and prostanoid synthesis modulating effects of Q13216 and FK-506 have been measured and compared with the immunosuppressive action of these drugs . Both , Q13216 and FK-506 inhibited proliferation of MC at similar doses ( IC50 approximately 1 microgram.ml-1 ) . Lymphoproliferation was suppressed with IC50s of 50 ng.ml-1 and < 1 ng.ml-1 , respectively . In contrast , and unlike FK-506 , Q13216 caused mesangiolysis ( IC50 = 4.5 micrograms.ml-1 ) and concentration dependently inhibited the interleukin-1 beta ( P01584 ) stimulated mesangial cell release of TXB2 at nanomolar doses ( IC50 = 50 ng.ml-1 ) . In kinetic experiments ( 6-48 h ) , Q13216 1 ng.ml-1 partially and 1 microgram.ml-1 completely abolished the P01584 augmented mesangial secretion TXB2 at all the time points tested . Both , low and high doses of Q13216 reduced DB00917 release by only 20-40 % and then not until at least 24 h of incubation . Measuring enzymatic capacity of membrane fractions of MC to generate TXB2 or DB00917 from added arachidonic acid ( 10(-5) M ) , Q13216 ( 0.1-1000 ng.ml-1 ) caused a dose dependent reduction in cyclooxygenase ( P36551 ) /thromboxane synthase activity up to 76 % , while DB00917 synthesis ( P36551 /prostaglandin synthase ) was decreased by 34 % . Immunoblots with a specific P23219 antiserum revealed that P23219 protein expression of MC was not affected by Q13216 . ( ABSTRACT TRUNCATED AT 250 WORDS ) Ablation of cholesterol biosynthesis in neural stem cells increases their P15692 expression and angiogenesis but causes neuron apoptosis . Although sufficient cholesterol supply is known to be crucial for neurons in the developing mammalian brain , the cholesterol requirement of neural stem and progenitor cells in the embryonic central nervous system has not been addressed . Here we have conditionally ablated the activity of squalene synthase ( P37268 ) , a key enzyme for endogenous cholesterol production , in the neural stem and progenitor cells of the ventricular zone ( VZ ) of the embryonic mouse brain . Mutant embryos exhibited a reduced brain size due to the atrophy of the neuronal layers , and died at birth . Analyses of the E11.5-E15.5 dorsal telencephalon and diencephalon revealed that this atrophy was due to massive apoptosis of newborn neurons , implying that this progeny of the P37268 -ablated neural stem and progenitor cells was dependent on endogenous cholesterol biosynthesis for survival . Interestingly , the neural stem and progenitor cells of the VZ , the primary target of P37268 inactivation , did not undergo significant apoptosis . Instead , vascular endothelial growth factor ( P15692 ) expression in these cells was strongly upregulated via a hypoxia-inducible factor-1-independent pathway , and angiogenesis in the VZ was increased . Consistent with an increased supply of lipoproteins to these cells , the level of lipid droplets containing triacylglycerides with unsaturated fatty acyl chains was found to be elevated . Our study establishes a direct link between intracellular cholesterol levels , P15692 expression , and angiogenesis . Moreover , our data reveal a hitherto unknown compensatory process by which the neural stem and progenitor cells of the developing mammalian brain evade the detrimental consequences of impaired endogenous cholesterol biosynthesis . Lymphagenesis correlates with expression of vascular endothelial growth factor-C in colorectal cancer . Lymphagenesis in gastrointestinal tumors is not well described . To clarify its presence and regulation , we assessed the microlymphatic count ( MLC ) in colorectal cancer patients . Lymphatic vessels were evaluated by enzyme-histochemistry for 5'-nucleotidase ( 5'-NA ) . Since vascular endothelial growth factor ( P15692 ) -C is reportedly associated with lymphagenesis , the expression of P49767 protein was immunohistochemically assessed by the catalyzed signal amplification ( Q13216 ) method . MLC of peritumoral lesions was significantly higher than that of non-cancer and intratumoral lesions ( p < 0.01 ) ; it increased where P49767 was highly expressed ( p < 0.01 ) and increased with the depth of invasion in peritumoral lesions . These results indicate significant findings at peritumoral lesion : that lymphagenesis may be elicited by tumor spread ; that P49767 expression is associated with lymphagenesis and is a potent factor stimulating lymphagenesis . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Inhibition of cyclooxygenase-2 elicits a P21554 -mediated decrease of excitatory transmission in rat P00915 hippocampus . Cannabinoid receptor ( P21554 ) ligands decrease excitatory and inhibitory transmission in the hippocampus , but the influence of endogenously formed cannabinoids ( eCBs ) on basal excitatory transmission remains uncertain . Here , we investigated the influence of eCBs on synaptic transmission in P00915 hippocampus using the slice preparation . Blockade of P21554 with the selective receptor antagonists SR141716 ( rimonabant ) or AM251 augmented synaptic responses evoked upon stimulation of the Schaffer collaterals . This effect persisted in the presence of bicuculline or CGP55845 to block GABA(A) or GABA(B) receptors , revealing a tonic eCB influence on excitatory transmission . Selective inhibition of cyclooxygenase-2 ( P35354 ) with meloxicam or NS-398 decreased excitatory responses partly in a P21554 -dependent manner , independently of GABA(A) transmission . Paired-pulse paradigms suggested a presynaptic P21554 mechanism to decrease glutamate release . Inhibition of P23219 or other routes of eCB degradation did not affect synaptic transmission . We conclude that P35354 regulates the formation of P21554 ligands that decrease hippocampal excitatory transmission . Trichostatin A , a histone deacetylase inhibitor , reverses epithelial-mesenchymal transition in colorectal cancer SW480 and prostate cancer PC3 cells . Trichostatin A ( P32119 ) is a kind of classical histone deacetylase ( HDAC ) inhibitor . In this study , we reported the reversal effects of P32119 on EMT and investigated the possible involved molecular mechanisms in SW480 and PC3 cells . Firstly , we observed that P32119 induced the reversal process of epithelial-mesenchymal transition ( EMT ) in SW480 and PC3 cells , resulting in attenuated cell invasion and migration abilities . P32119 -induced EMT reversal was characterized by up-regulation of P12830 and down-regulation of P08670 . Then , treatment with P32119 also decreased the expression of transcription factor Slug . Furthermore , over-expression of Slug significantly caused down-regulation of P12830 and up-regulation of P08670 . Meanwhile , P32119 treatment in Slug-expressing cells could prevent these changes . These findings suggested that Slug played a crucial role in P32119 -induced EMT reversal . Additionally , the study showed that P32119 could induce the increase of Q13547 and Q92769 on the Slug gene promoter , which might be responsible for the suppression of Slug . Overall , P32119 could reverse EMT in SW480 and PC3 cells and P32119 -mediated down-regulation of Slug was involved in the reversal process . Celecoxib offsets the negative renal influences of cyclosporine via modulation of the TGF-β1/ P60568 / P35354 /endothelin ET(B) receptor cascade . Endothelin ( ET ) signaling provokes nephrotoxicity induced by the immunosuppressant drug cyclosporine A ( Q13216 ) . We tested the hypotheses that ( i ) : celecoxib , a selective cyclooxygenase-2 ( P35354 ) inhibitor , counterbalances renal derangements caused by Q13216 in rats and ( ii ) the P35354 /endothelin ET(B) receptor signaling mediates the Q13216 -celecoxib interaction . Ten-day treatment with Q13216 ( 20 mg/kg/day ) significantly increased biochemical indices of renal function ( serum urea , creatinine ) , inflammation ( interleukin-2 , P60568 ) and fibrosis ( transforming growth factor-β₁ , TGF-β₁ ) . Histologically , Q13216 caused renal tubular atrophy along with interstitial fibrosis . These detrimental renal effects of Q13216 were largely reduced in rats treated concurrently with celecoxib ( 10 mg/kg/day ) . We also report that cortical glomerular and medullary tubular protein expressions of P35354 and ET(B) receptors were reduced by Q13216 and restored to near-control values in rats treated simultaneously with celecoxib . The importance of ET(B) receptors in renal control and in the Q13216 -celecoxib interaction was further verified by the findings ( i ) most of the adverse biochemical , inflammatory , and histopathological profiles of Q13216 were replicated in rats treated with the endothelin ETB receptor antagonist BQ788 ( 0.1 mg/kg/day , 10 days ) , and ( ii ) the BQ788 effects , like those of Q13216 , were alleviated in rats treated concurrently with celecoxib . Together , the data suggest that the facilitation of the interplay between the TGF-β1/ P60568 / P35354 pathway and the endothelin ET(B) receptors constitutes the cellular mechanism by which celecoxib ameliorates the nephrotoxic manifestations of Q13216 in rats . DB00328 ameliorates high glucose-induced proliferation and invasion via modulation of e-cadherin in pancreatic cancer cells . DB00328 , an inhibitor of cyclooxygenase-2 ( P35354 ) , has been shown to exert anticancer effects in a variety of cancers . However , the effect and mechanism of indometacin on high glucose ( HG ) -induced proliferation and invasion of pancreatic cancer ( PC ) cells remain unclear . Multiple lines of evidence suggest that a large portion of pancreatic cancer ( PC ) patients suffer from either diabetes or HG which contributing PC progression . In this study , we report that indometacin down-regulated HG-induced proliferation and invasion via up-regulating P12830 but not P35354 in PC cells . Additionally , the P12830 transcriptional repressors , Snail and Slug , were also involved in the process . Furthermore , the proliferation and invasion of PC cells , incubated in HG medium and treated with indometacin were significantly increased when P12830 was knocked down ( Si-E-cad ) . Moreover , the protein levels of P08253 , P14780 , and P15692 were increased in PC cells transfected with Si-E-cad . Finally , the activation of the PI3K/AKT/GSK-3β signaling pathway was demonstrated to be involved in indometacin reversing HG-induced cell proliferation and invasion in PC cells . In conclusion , these results suggest that indometacin plays a key role in down-regulating HG-induced proliferation and invasion in PC cells . Our findings indicate that indometacin could be used as a novel therapeutic strategy to treat PC patients who simultaneously suffer from diabetes or HG . DB11320 modulates multiple functional activities of monocyte-derived dendritic cell subsets via histamine receptor 2 . Expression of CD1a proteins in human monocyte-derived dendritic cells ( DCs ) specifies functionally distinct subsets with different inflammatory properties . DB11320 is recognized as an inflammatory mediator released by various cell types including DCs . The diverse biological effects of histamine are mediated by G-protein-coupled histamine receptors ( HRs ) , which are able to modulate the functional activities of DC subsets . The goal of the present study was to compare the expression and activity of HRs in the CD1a(-) and CD1a(+) monocyte-derived DC subsets and to test the effects of histamine on the differentiation , activation and functional activities of these subsets . We show that P25021 is present at high levels in both DC subsets , whereas P35367 and Q9H3N8 are expressed in a subset-specific manner . DB11320 shifts DC differentiation to the development of CD1a(-) DCs and modulates DC activation through its inhibitory effect on CD1a(+) DC differentiation . DB11320 -induced reduction of CD1a(+) DCs is associated with increased secretion of P05231 and P22301 , up-regulation of a typical combination of chemokines , expression C5aR1 by the CD1a(-) DC subset and enhanced migration of both activated DC subsets supported by the production of P14780 and P39900 enzymes . All these effects were shown to be mediated in a P25021 -specific manner as revealed by the specific antagonist of the receptor . As P25021 is expressed at high levels in both DC subsets , we propose that it may dominate the regulation of multiple DC functions . In contrast , P35367 and Q9H3N8 with opposing subset-related expression may have a regulatory or fine-tuning role in histamine-induced functional activities . Eupolyphaga sinensis walker displays inhibition on hepatocellular carcinoma through regulating cell growth and metastasis signaling . Tumor growth and metastasis are responsible for most cancer patients ' deaths . Here , we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis . Compared with proliferation , colony formation , transwell assay and transplantable tumor in nude mouse in vitro and vivo , eupolyphaga sinensis walker extract ( ESWE ) showed good inhibition on the SMMC-7721 cell growth and metastasis . Using genome-wide microarray analysis , we found the down-regulated growth and metastasis factors , and selected down-regulated genes were confirmed by real-time PCR . Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE . Moreover , our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ , Akt , m-TOR , Erk1/2 , MEK-2 , Raf and JNK-1 , but also increasing cyclin D1 protein levels and decreasing amount of cyclin E , cyclin B1 and cdc2 of the cycle proteins . At the same time , ESWE reduced P08253 , P14780 and P61073 , P00747 , NFκB and P04637 activities . Overall , our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC , AKT , MAPK signaling and related metastasis signaling , having potential in cancer therapy . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . The protective effect of rebamipide on paracellular permeability of rat gastric epithelial cells . BACKGROUND : Barrier function in gastric epithelial cells is essential for the gastric defence mechanism against acid back-diffusion into the mucosal layer . Our previous study indicated that trans-epithelial resistance ( Q9NZ01 ) of rat gastric epithelial cells was rapidly increased when the cells were exposed to acid . This response to acid was diminished by indometacin . AIM : Evaluate the effects of a mucoprotective agent , rebamipide , on the nonsteroidal anti-inflammatory drug ( NSAID ) -induced increase of gastric epithelial permeability . METHODS : Rat gastric epithelial cells were plated on tissue culture inserts . Cells were exposed to a NSAID ( indometacin , 10-7 M ) . Trans-epithelial permeability was measured by Q9NZ01 and diffusion rate of 14C-mannitol . The effect of rebamipide was evaluated by measuring Q9NZ01 . Endogenous prostaglandin E2 ( DB00917 ) production in culture medium was also measured . RESULTS : DB00328 gradually and significantly decreased Q9NZ01 and increased 14C-manitol permeability . Rebamipide reversed the indometacin-induced changes in epithelial permeability and induced DB00917 synthesis . This induction was blocked by either indometacin or a Cyclooxygenase ( P36551 ) -2 specific inhibitor . CONCLUSIONS : P36551 inhibitors such as indometacin inhibit regulation of epithelial permeability by reducing DB00917 . P23219 has an important role in the gastric defense mechanism . Rebamipide suppressed an indometacin-induced increase in gastric epithelial permeability by increasing DB00917 levels in a P35354 dependent manner . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . Genetic mechanism of aspirin-induced urticaria/angioedema . PURPOSE OF REVIEW : DB00945 -induced urticaria/angioedema is a major aspirin-related hypersensitivity often associated with aspirin-intolerant asthma . Genetic studies on aspirin-intolerant asthma have shown chronic overproduction of cysteinyl leukotrienes . The genetic analysis of aspirin-induced urticaria/angioedema is limited , however . RECENT FINDINGS : A recent study on HLA genotypes has suggested that the HLA alleles DRB11302 and DQB10609 may be genetic markers for aspirin-induced urticaria/angioedema . A polymorphism study that examined nine single-nucleotide polymorphisms of five leukotriene-related genes [ P09917 ( encoding P09917 ) , P20292 ( P09917 -activating protein ) , P35354 ( cyclooxygenase 2 ) , Q16873 ( leukotriene C4 synthase ) , and Q9Y271 ( cysteinyl leukotriene receptor 1 ) ] found that promoter polymorphisms of P09917 ( -1708A > G ) and Q9Y271 ( -634C > T ) were significantly different between aspirin-intolerant asthma and aspirin-induced urticaria/angioedema , suggesting different contributions to the lipoxygenase pathway . A second polymorphism study , conducted on histamine-related genes , did not find any significant associations with aspirin-induced urticaria/angioedema for the genes P50135 ( encoding histamine N-methyltransferase ) , P35367 or P25021 ( encoding histamine receptor types 1 and 2 respectively ) , or the gene encoding high-affinity IgE receptor Ibeta ( FcepsilonRIbeta ) ; however , the FcepsilonRIalpha gene promoter polymorphism was significantly associated with aspirin-induced urticaria/angioedema . This finding has been supported by in vitro functional studies . SUMMARY : The HLA alleles DRB11302 and DQB10609 , and the P09917 and FcepsilonRIalpha promoter polymorphisms , may contribute to the pathogenesis of aspirin-induced urticaria/angioedema . Further investigation to identify candidate genetic markers would help to elucidate the pathogenic mechanism of this condition . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion .

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n-3 fatty acids specifically modulate catabolic factors involved in articular cartilage degradation . This study describes specific molecular mechanisms by which supplementation with n-3 fatty acids ( i.e. those present in fish oils ) can modulate the expression and activity of degradative and inflammatory factors that cause cartilage destruction during arthritis . Our data show that incorporation of n-3 fatty acids ( but not other polyunsaturated or saturated fatty acids ) into articular cartilage chondrocyte membranes results in a dose-dependent reduction in : ( i ) the expression and activity of proteoglycan degrading enzymes ( aggrecanases ) and ( ii ) the expression of inflammation-inducible cytokines ( interleukin ( IL ) -1alpha and tumor necrosis factor ( P01375 ) -alpha ) and cyclooxygenase ( P35354 ) , but not the constitutively expressed cyclooxygenase P23219 . These findings provide evidence that n-3 fatty acid supplementation can specifically affect regulatory mechanisms involved in chondrocyte gene transcription and thus further advocate a beneficial role for dietary fish oil supplementation in alleviation of several of the physiological parameters that cause and propogate arthritic disease . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Angiotensin II regulates V2 receptor and pAQP2 during ureteral obstruction . Release of bilateral ureteral obstruction ( BUO ) is associated with nephrogenic diabetes insipidus ( NDI ) and a reduced abundance of the vasopressin-regulated aquaporins . To evaluate the role of the vasopressin type 2 receptor ( P30518 ) , we determined P30518 abundance in kidneys from rats subjected to 24-h BUO or 24-h unilateral ureteral obstruction ( UUO ) followed by 48-h release . Because angiotensin II type 1 ( AT1 ) receptor blockade attenuates postobstructive polyuria and aquaporin-2 ( P41181 ) downregulation , we examined the effect of AT1 receptor blockade on P41181 phosphorylated at serine 256 ( pS256- P41181 ) and V2 receptor complex abundance in kidney inner medulla ( IM ) . Furthermore , DB02527 generation in sodium fluoride- and forskolin-stimulated inner medullary membrane fractions was studied after release of BUO . P30518 was significantly reduced to 12 % of sham levels in IM and to 52 % of sham levels in cortex and outer stripe of outer medulla ( OSOM ) from BUO rats . In UUO rats , P30518 abundance in the obstructed kidney IM decreased to 35 % of sham levels , whereas it was comparable to sham levels in the nonobstructed kidney IM . No significant change was observed in cortex and OSOM . AT1 receptor blockade attenuated P30518 , pS256- P41181 , and G(s)alpha protein downregulation in IM and partially reversed the obstruction-induced inhibition of sodium fluoride- and forskolin-stimulated DB02527 generation in inner medullary membrane fractions from BUO rats . In conclusion , P30518 downregulation plays a pivotal role in development of NDI after release of BUO . In addition , we have shown that angiotensin II regulates the V2 receptor complex and pS256- P41181 in postobstructive kidney IM , probably by stimulating DB02527 generation . A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . Lentiviral infection of rhesus macaques causes long-term injury to cortical and hippocampal projections of prostaglandin-expressing cholinergic basal forebrain neurons . The simian immunodeficiency virus ( SIV ) macaque model resembles human immunodeficiency virus-acquired immunodeficiency syndrome ( AIDS ) and associated brain dysfunction . Altered expression of synaptic markers and transmitters in neuro-AIDS has been reported , but limited data exist for the cholinergic system and lipid mediators such as prostaglandins . Here , we analyzed cholinergic basal forebrain neurons with their telencephalic projections and the rate-limiting enzymes for prostaglandin synthesis , cyclooxygenase isotypes 1 and 2 ( P23219 and P35354 ) in the brains of SIV-infected macaques with or without encephalitis and antiretroviral therapy and uninfected controls.Cyclooxygenase isotype 1 , but not P35354 , was coexpressed with markers of cholinergic phenotype , that is , choline acetyltransferase and vesicular acetylcholine transporter ( Q16572 ) , in basal forebrain neurons of monkey , as well as human , brain . Cyclooxygenase isotype 1 was decreased in basal forebrain neurons in macaques with AIDS versus uninfected and asymptomatic SIV-infected macaques . The Q16572 -positive fiber density was reduced in frontal , parietal , and hippocampal-entorhinal cortex . Although brain SIV burden and associated P23219 - and P35354 -positive mononuclear and endothelial inflammatory reactions were mostly reversed in AIDS-diseased macaques that received 6-chloro-2',3'-dideoxyguanosine treatment , decreased Q16572 -positive terminal density and reduced cholinergic P23219 expression were not . Thus , P23219 expression is a feature of primate cholinergic basal forebrain neurons ; it may be functionally important and a critical biomarker of cholinergic dysregulation accompanying lentiviral encephalopathy . These results further imply that insufficiently prompt initiation of antiretroviral therapy in lentiviral infection may lead to neurostructurally unremarkable but neurochemically prominent irreversible brain damage . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . P01116 , P00533 , P09619 -α , P10721 and P35354 status in carcinoma showing thymus-like elements ( CASTLE ) . BACKGROUND : CASTLE ( Carcinoma showing thymus-like elements ) is a rare malignant neoplasm of the thyroid resembling lymphoepithelioma-like and squamous cell carcinoma of the thymus with different biological behaviour and a better prognosis than anaplastic carcinoma of the thyroid . METHODS : We retrospectively investigated 6 cases of this very rare neoplasm in order to investigate the mutational status of P01116 , P00533 , P09619 -α and P10721 , as well as the immunohistochemical expression pattern of CD117 , P00533 and P35354 , and possibly find new therapeutic targets . RESULTS : Diagnosis was confirmed by a moderate to strong expression of P06127 , CD117 and CK5/6 , whereas thyroglobulin , calcitonin and Q15669 -1 were negative in all cases . Tumors were also positive for P35354 and in nearly all cases for P00533 . In four cases single nucleotide polymorphisms ( SNPs ) could be detected in exon 12 of the P09619 -α gene ( rs1873778 ) , in three cases SNPs were found in exon 20 of the P00533 gene ( rs1050171 ) . No mutations were found in the P10721 and P01116 gene . CONCLUSIONS : All tumors showed a P35354 expression as well as an P00533 expression except for one case and a wild-type P01116 status . No activating mutations in the P00533 , P10721 and P09619 -α gene could be detected . Our data may indicate a potential for targeted therapies , but if these therapeutic strategies are of benefit in CASTLE remains to be determined . VIRTUAL SLIDES : The virtual slide(s) for this article can be found here : http://www.diagnosticpathology.diagnomx.eu/vs/1658499296115016 . Garcinone B reduces prostaglandin E2 release and NF-kappaB-mediated transcription in P13671 rat glioma cells . In the course of our survey of natural compounds inhibiting prostaglandin E2 release and/or lipopolysaccharide ( LPS ) -induced transcriptional stimulation via NF-kappaB , a central regulator of inflammatory genes , from natural resources , we found garcinone B , a xanthone from callus tissue culture of Hypericum patulum , as a compound with such pharmacological activities , that is a derivative of gamma-mangostin which potently inhibits P23219 and P35354 activities to reduce DB00917 release from P13671 rat glioma cells , and inhibits IKK activity to prevent NF-kappaB-dependent P35354 gene transcription . Garcinone B , to a lesser extent , reduced A23187-induced increase in prostaglandin E2 release than gamma-mangostin and its structurally related compound , patulone , in P13671 cells . This compound also prevented LPS-induced stimulation of NF-kappaB-dependent transcription . These results suggest that garcinone B becomes a unique pharmacological tool to investigate intracellular signaling pathways involved in inflammation . The effect of prenatal nicotine on mRNA of central cholinergic markers and hematological parameters in rat fetuses . A number of studies have demonstrated the influence of nicotine on fetal development . This study determined the expression of choline acetyltransferase ( P28329 ) , vesicular acetylcholine transporter ( Q16572 ) , and high-affinity choline transporter ( Q9GZV3 ) in the forebrain and hindbrain following chronic prenatal nicotine exposure in the rat fetus ( maternal rats were subcutaneously injected with nicotine at different gestation periods ) . We also measured the effect of chronic nicotine exposure on fetal blood pO(2) , pCO(2) , pH , Na(+) and K(+) concentrations , as well as lactic acid levels . Maternal nicotine exposure during pregnancy was associated with a decrease in fetal pO(2) coupled with a significant increase in pCO(2) and lactic acid as well as restricted fetal growth . Additionally , maternal nicotine administration also reduced P28329 , Q16572 , and Q9GZV3 mRNA levels in the fetal brain . DB00184 -induced fetal hypoxic responses and reduced cholinergic marker expression in the brain were more severe when nicotine was started in early gestation . Our results provide new information about the effects of repeated exposure to nicotine in utero on the expression of central P28329 , Q16572 , and Q9GZV3 in the rat fetus . These results indicate that repeated hypoxic episodes or/and a direct effect of nicotine on the central cholinergic system during pregnancy may contribute to brain developmental problems in fetal origin . DB00227 induces the expression of bradykinin type 2 receptors in cultured human coronary artery endothelial cells . Cardioprotective bradykinin type-2 receptors ( BK-2Rs ) are downregulated in the myocardial endothelium of both human and rat failing hearts . Statins are cardioprotective drugs that reduce the level of plasma cholesterol but also exert cholesterol-independent pleiotropic effects . Here we examined the effect of lovastatin on BK-2R expression in cultured human coronary artery endothelial cells . The effect of lovastatin on the expression of BK receptors in human coronary artery endothelial cells ( HCAECs ) was examined by real-time PCR , Western blot analysis and immunocytochemistry . DB00227 induced a time- and concentration-dependent increase in both BK-2R and BK-1R mRNA expression in the cultured HCAECs . Also , the number of functional BK-2Rs capable of inducing BK-mediated NO production and cGMP signaling was increased in the lovastatin-treated HCAECs . Mevalonate , the direct metabolite of P04035 , reversed the effect of lovastatin . Furthermore , lovastatin inhibited Rho activation and a selective inhibitor of Rho-associated kinases , Y-27632 , induced a similar increase in BK-2R expression as lovastatin . In contrast , a specific inhibitor of P35354 , NS398 , significantly inhibited the lovastatin-induced expression of BK-2Rs . Here we show for the first time that lovastatin induces the expression of BK-2Rs in cultured human coronary artery endothelial cells through a novel cholesterol-independent pleiotropic mechanism that involves RhoA kinase inhibition and P35354 activation . Thus , reported beneficial effects of statins in cardiovascular diseases may be partly mediated by an increased expression of cardioprotective BK-2Rs in the endothelial cells of the coronary tree . Moreover , the use of P35354 inhibitors may affect the level of endothelial BK-2Rs in a negative fashion . Meloxicam . Meloxicam ( DB00814 trade mark , Boehringer Ingelheim ) is a relatively new oral non-steroidal anti-inflammatory drug ( NSAID ) approved for the treatment of osteoarthritis in the US . It has also been evaluated for the treatment of rheumatoid arthritis , ankylosing spondylitis and acute ' rheumatic ' pain . Meloxicam has been shown to be P35354 preferential , particularly at its lowest therapeutic dose , and is anti-inflammatory by inhibiting prostanoid synthesis in inflammatory cells . Since it is P35354 preferential , it would be expected to have less gastrointestinal toxicity than non-selective NSAIDs . In clinical trials of meloxicam in osteoarthritis , it was found to be as effective as piroxicam , diclofenac and naproxen with less clinical gastrointestinal symptoms and less perforations , obstructions and bleeds by meta-analysis . Adverse events , including peripheral oedema and hypertension , occurred at a similar rate as with traditional NSAIDs . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Disruption of cyclooxygenase-2 prevents downregulation of cortical P41181 and Q92482 in response to bilateral ureteral obstruction in the mouse . Bilateral ureteral obstruction ( BUO ) in rats is associated with increased cyclooxygenase type 2 ( P35354 ) expression , and selective P35354 inhibition prevents downregulation of aquaporins ( AQPs ) in response to BUO . It was hypothesized that a murine model would display similar changes in renal P35354 and AQPs upon BUO and that targeted disruption of P35354 protects against BUO-induced suppression of collecting duct AQPs . P35354 (-/-) and wild-type littermates ( C57BL/6 ) were employed to determine P23219 , -2 , P41181 , and Q92482 protein abundances and localization after BUO . In a separate series , sham and BUO wild-type mice were treated with a selective P35354 inhibitor , parecoxib . The P35354 protein level increased in wild-type mice in response to BUO and was not detectable in P35354 (-/-) . P23219 protein abundance was increased in sham-operated and BUO mice . Total P41181 and -3 mRNA and protein levels decreased significantly after BUO in the cortex+outer medulla ( C+OM ) and inner medulla ( IM ) . The decrease in C+OM P41181 and -3 levels was attenuated/prevented in P35354 (-/-) mice , whereas there was no change in the IM . In parallel , inhibition of P35354 by parecoxib rescued C+OM Q92482 and IM P41181 protein level in wild-type mice subjected to BUO . In summary , 1 ) In C57BL/6 mice , ureteral obstruction increases renal P35354 expression in interstitial cells and lowers P41181 /-3 abundance and 2 ) inhibition of P35354 activity by targeted disruption or pharmacological blockade attenuates obstruction-induced AQP downregulation . In conclusion , P35354 -derived prostaglandins contribute to downregulation of transcellular water transporters in the collecting duct and likely to postobstruction diureses in the mouse . Thrombin inhibits migration of human hepatic myofibroblasts . Several lines of data recently pointed out a role of the serine proteinase thrombin in liver fibrogenesis , but its mechanism of action is unknown . The aim of this study was to evaluate the effect of thrombin on the migration of human liver myofibroblasts . We show here that thrombin inhibits both basal migration and platelet-derived growth factor ( PDGF ) -BB-induced migration of myofibroblasts . By using a thrombin antagonist , a protease-activated receptor ( PAR ) -1 mimetic peptide , and a P25116 antibody , we show that this effect is dependent on the catalytic activity of thrombin and on P25116 activation . Thrombin 's effect on basal migration was dependent on cyclooxygenase 2 ( P35354 ) activation because it was blocked by the P35354 inhibitors NS-398 and nimesulide , and pharmacological studies showed that it was relayed through prostaglandin E(2) and its EP(2) receptor . On the other hand , thrombin-induced inhibition of DB00102 -induced migration was not dependent on P35354 . We show that thrombin inhibits PDGF-induced Akt-1 phosphorylation . This effect was consecutive to inhibition of PDGF-beta receptor activation through active dephosphorylation . Thus thrombin , through two distinct mechanisms , inhibits both basal- and DB00102 -induced migration of human hepatic liver myofibroblasts . The fine tuning of myofibroblast migration may be one of the mechanisms used by thrombin to regulate liver fibrogenesis . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity .

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Purinergic receptors involved in the immunomodulatory effects of DB00171 in human blood . We recently showed that the physiological compound DB00171 simultaneously inhibited P01375 and stimulated P22301 release in LPS-PHA stimulated blood . The purpose of the present study was to determine the mechanism involved in the concerted modulatory effect of DB00171 on P01375 and P22301 . Incubation of blood with DB00171 in the presence of selective P2 receptor antagonists showed that the stimulatory effect of DB00171 on P22301 release was completely annihilated by both 2-MeSAMP ( a Q9H244 /13 receptor antagonist ) and PSB-0413 ( a Q9H244 receptor antagonist ) . On the other hand , the inhibitory effect of DB00171 on P01375 release was completely reversed by 5'- P30566 ( a Q96G91 receptor antagonist ) as well as by H-89 , an inhibitor of DB02527 -activated PKA . The concerted inhibition by DB00171 of P01375 release via Q96G91 activation and stimulation of P22301 release via Q9H244 activation implicates a novel approach towards immunomodulation by altering the balance among pro- and anti-inflammatory cytokines . Serum deprivation confers the MDA-MB-231 breast cancer line with an P00533 / P52333 / O14939 system that maximizes cancer cell invasion . Our laboratory has reported earlier that in leukocytes , phospholipase D2 ( O14939 ) is under control of P52333 ( P52333 ) , which mediates chemotaxis . Investigating P52333 in cancer cells led to an important discovery as exponentially growing MDA-MB-231 human breast cancer cells , which are highly proliferative and metastatic , did not substantially use P52333 to activate O14939 . However , in 2-h or 16-h starved cell cultures , P52333 switches to a O14939 -enhancing role , consistent with the needs of those cells to enter a " survival state " that relies on an increase in O14939 activity to withstand serum deprivation . Using a small-molecule tyrosine kinase inhibitor , the flavonoid 4',5,7-trihydroxyflavone ( apigenin ) , as well as RNA silencing , we found that the invasive phenotype of MDA-MB-231 cells is mediated by O14939 under direct regulation of both P52333 and the tyrosine kinase , epidermal growth factor receptor ( P00533 ) . Furthermore , serum-deprived cells in culture show an upregulated P00533 / P52333 / O14939 -PA system and are especially sensitive to a combination of P52333 and O14939 enzymatic activity inhibitors ( 30nM apigenin and 300nM 5-fluoro-2-indolyl des-chlorohalopemide ( FIPI ) , respectively ) . Thus , a multi-layered activation of cell invasion by two kinases ( P00533 and P52333 ) and a phospholipase ( O14939 ) provides regulatory flexibility and maximizes the aggressively invasive power of MDA-MB-231 breast cancer cells . This is especially important in the absence of growth factors in serum , coincidental with migration of these cells to new locations . Janus kinase inhibition with tofacitinib : changing the face of inflammatory bowel disease treatment . The advent of anti-Tumor Necrosis Factor ( P01375 ) therapy has changed the way of treating inflammatory bowel disease ( Q9UKU7 ) . However , primary and secondary failure are relatively frequent with all anti- P01375 agents , which are available only as parenteral agents . DB08895 is an oral janus kinase ( JAK ) inhibitor that inhibits JAK family kinase members , in particular P23458 and P52333 , achieving a broad limitation of inflammation by interfering with several cytokine receptors . It first proved its efficacy as an immunosuppressive regimen after renal transplantation , and was recently approved by the FDA for rheumatoid arthritis . First data in Q9UKU7 are promising , especially in ulcerative colitis . Ongoing clinical trials in both UC and Crohn 's disease ( CD ) are needed to further explore its efficacy in CD and to better assess its safety profile . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . [ The effect of leptin and its mechanisms on the migration and invasion of human breast cancer MCF-7 cells ] . OBJECTIVE : To investigate the effect and the relevant molecular mechanisms of leptin on the migration and invasion of human breast cancer MCF-7 cells . METHODS : The expression of P48357 in MCF-7 cells was measured by RT-PCR and Western blotting . The effects of leptin ( 100 ng/mL ) on the the phosphorylation of a few key cell signaling proteins , p- P27361 /2 , p- P40763 , p-AKT in MCF-7 cells were examined by Western blotting . Cell scratch assay and Transwell(TM) ; assay were utilized to measure the effects of leptin on the migration and invasion capability of MCF-7 cells , respectively . The effects of leptin on the mRNA and protein expression of matrix metalloproteinas 9 ( P14780 ) and transforming growth factor β ( TGF-β ) were measured by RT-PCR and Western blotting . RESULTS : Both OB-Rb and OB-Rt were expressed in MCF-7 cells . This indicated that leptin may have significant activities in MCF7 cells . Indeed , leptin increased the phosphorylation of p- P27361 /2 , p- P40763 , and p-AKT in MCF-7 cells ( P < 0.05 ) . Further , leptin promoted migration and invasion of MCF-7 cells , which were attenuated by the JAK/ P35610 inhibitor AG490 ( 50 μmol/L ) , and the PI3K/AKT inhibitor LY294002 ( 10 μmol/L ) ( P < 0.05 ) . Similarly , leptin also increased the mRNA and protein expression of P14780 and TGF-β , and these effects were blocked by AG490 and LY294002 as well ( P < 0.05 ) . CONCLUSION : Leptin promoted the migration and invasion capabilities of MCF-7 cells . These activities may be achieved by the upregulation of P14780 and TGF-β through JAK/ P35610 and PI3K/AKT signaling pathways . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . JAK- P35610 pathway and myogenic differentiation . Myogenic differentiation plays an important role in muscle regeneration and is regulated by two transcription factor families , MRFs and Q02078 , which induce differentiation of myoblasts through expression of the muscle-specific gene , myogenin . In addition , many intracellular signaling pathways are also involved in myogenic differentiation , including p38 MAPK , P29323 /MAPK and PI3K/AKT . The JAK- P35610 pathway is activated by various cytokines and positively or negatively regulates the differentiation of myoblasts . P23458 plays a notable role in proliferation ; whereas , O60674 and P52333 function mainly in differentiation . The STATs , molecules downstream of JAK , regulate myogenesis . With P23458 , P42224 promotes proliferation , while P40763 has a dual effect on proliferation and differentiation . The JAK- P35610 negative regulator , Q9NSE2 , is also associated with myogenesis ; although , its role is controversial . In this review , we will discuss the role of the JAK- P35610 pathway on myogenic differentiation . Clinical utility of the oral JAK inhibitor tofacitinib in the treatment of rheumatoid arthritis . Immune/inflammatory cells act in rheumatoid arthritis ( RA ) -affected patients by synthesizing several inflammatory mediators , including cytokines that initiate intracellular signaling . Recently , small molecule inhibitors of transduction and transcription signals that influence the intracellular pathways ( such as the Janus kinase [ JAK ] family of tyrosine kinases ) have been tested for RA treatment . Four members of the JAK family are known : P23458 , O60674 , P52333 , and TyK2 . P23458 / P52333 constitutively binds to the cytoplasmic portion of the cytokine receptor - the common gamma chain - that represents a common subunit of several cytokines involved in T-cell and natural killer cell development , as well as in B-cell activation . DB08895 is an oral JAK inhibitor that is now available and effective in RA treatment , as shown in multiple Phase II and Phase III clinical trials . However , long-term safety data and comparisons with other disease-modifying antirheumatic drugs and small molecule inhibitors are necessary to better determine the role of tofacitinib in RA . Indirubin inhibits tumor growth by antitumor angiogenesis via blocking P35968 -mediated JAK/ P40763 signaling in endothelial cell . Tumor angiogenesis is one of the hallmarks of the development in malignant neoplasias and metastasis . Many angiogenesis inhibitors are small molecules from natural products . Indirubin , the active component of a traditional Chinese herbal medicine , Banlangen , has been shown to exhibit antitumor and anti-inflammation effects . But its roles in tumor angiogenesis , the key step involved in tumor growth and metastasis , and the involved molecular mechanism is unknown . Here , we identified that indirubin inhibited prostate tumor growth through inhibiting tumor angiogenesis . Using chick chorioallantoic membrane ( P62158 ) assay and mouse corneal model , we found that indirubin inhibited angiogenesis in vivo . We also showed the inhibition activity of indirubin in endothelial cell migration , tube formation and cell survival in vitro . Furthermore , indirubin suppressed vascular endothelial growth factor receptor 2-mediated Janus kinase ( JAK ) / P40763 signaling pathway but had little effects on the activity of extracellular signal-regulated kinase ( P29323 ) and p38 mitogen-activated protein kinase in endothelial cell . Our study provided the first evidence for antitumor angiogenesis activity of indirubin and the related molecular mechanism . Our investigations suggested that indirubin was a potential drug candidate for angiogenesis related diseases . Novel treatments with small molecules in psoriatic arthritis . Current treatment options for patients with active psoriatic arthritis ( PsA ) include synthetic disease-modifying antirheumatic drugs and biologic agents . Propelled by increased understanding of immunopathogenesis of PsA , new therapeutic agents targeting different biologic pathways have been evaluated . This article discusses novel small-molecule , orally available treatments that are currently in clinical development for the treatment of psoriasis and PsA . This includes the phosphodiesterase 4 inhibitor apremilast and Janus kinase ( JAK ) inhibitors . Apremilast has demonstrated significant improvements in patients with moderate to severe psoriasis and PsA in phase II and III clinical trials and has recently been approved for the treatment of PsA . DB08895 , an oral inhibitor of P52333 , P23458 , and , to a lesser degree , O60674 , approved for the treatment of rheumatoid arthritis in several countries , has demonstrated positive results in psoriasis in phase II studies . Studies in PsA are ongoing . With these new developments , treatment options will continue to improve in the future . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . c-Fes tyrosine kinase binds to and activates P40763 after granulocyte-macrophage colony-stimulating factor stimulation . Granulocyte-macrophage colony stimulating factor ( GM- P04141 ) induces proliferation and maturation of myeloid progenitor cells and also activates neutrophils . In order to investigate the pleiotropic effects of GM- P04141 stimulation , we examined the signaling pathways of protein tyrosine kinases ( PTKs ) and signal transducers and activators of transcription ( STATs ) in GM- P04141 -dependent proliferation of leukemia cells . Using TF-1 , a GM- P04141 -dependent human erythroleukemia cell line , we found that GM- P04141 enhanced DNA-binding and tyrosine phosphorylation of P40763 . GM- P04141 receptor ( GM-CSFR ) and c-Fes tyrosine kinase were also activated upon GM- P04141 stimulation . Furthermore , c-Fes formed a complex with P40763 . Experiments using a c-Fes mutant that lacked tyrosine kinase activity revealed that the activation of P40763 is kinase-dependent , but that the c-Fes- P40763 interaction is not affected by c-Fes tyrosine kinase activity . The results suggest that P40763 is activated by c-Fes tyrosine kinase through direct interaction during hematopoietic cell proliferation induced by GM- P04141 . Transforming properties of chimeric P41212 -JAK proteins in Ba/ P13726 cells . The involvement of the cytokine signaling pathway in oncogenesis has long been postulated . Recently , rearrangements of the gene encoding the tyrosine O60674 ( O60674 ) have been reported in human leukemias indicating a direct JAK-signal transduction and activator of transcription ( P35610 ) -mediated leukemic process . The leukemia-associated P41212 - O60674 fusion protein is formed by the oligomerization domain of the translocated ets leukemia ( P41212 ) protein fused to the catalytic domain of O60674 . P41212 -mediated oligomerization results in a constitutive tyrosine kinase activity that , in turn , is able to confer growth factor independence to the murine hematopoietic interleukin-3 ( P08700 ) -dependent Ba/ P13726 cell line . Results of the present study indicate that fusion proteins containing the oligomerization domain of P41212 and the tyrosine kinase domains of Jak1 , Jak2 , P52333 , or P29597 share similar properties and are able to efficiently substitute for the survival and mitogenic signals controlled by P08700 , without concomitant activation of the P08700 receptor . Electrophoretic mobility shift assays demonstrated Stat5 as the only activated Stat factor in P41212 -Jak2- and P41212 -Jak1-expressing cells , whereas other Stats , namely Stat1 and Stat3 , could be detected in P41212 - P52333 - , P41212 - P29597 - , and also in P41212 - P00519 -expressing Ba/ P13726 cells . High levels of expression of the Stat5-target genes pim-1 , osm , and Cis were observed in all the cytokine-independent cell lines . Furthermore , the expression of a dominant negative form of Stat5A markedly interfered with the growth factor independence process mediated by P41212 -Jak2 in Ba/ P13726 cells . Because the P11274 - P00519 and P41212 -PDGFbetaR oncoproteins also activate Stat5 , activation of this factor should be a crucial step in activated tyrosine kinase-mediated leukemogenesis . ( Blood. 2000;95:2076-2083 ) [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . 18 Beta-glycyrrhetinic acid triggers curative Th1 response and nitric oxide up-regulation in experimental visceral leishmaniasis associated with the activation of NF-kappa B . The efficacy of 18beta-glycyrrhetinic acid ( GRA ) , a pentacyclic triterpene belonging to the beta-amyrin series of plant origin , was evaluated in experimental visceral leishmaniasis . GRA is reported to have antitumor and immunoregulatory activities , which may be attributable in part to the induction of NO . Indeed , an 11-fold increase in NO production was observed with 20 microM GRA in mouse peritoneal macrophages infected with Leishmania donovani promastigotes . In addition to having appreciable inhibitory effects on amastigote multiplication within macrophages ( IC(50) , 4.6 microg/ml ) , complete elimination of liver and spleen parasite burden was achieved by GRA at a dose of 50 mg/kg/day , given three times , 5 days apart , in a 45-day mouse model of visceral leishmaniasis . GRA treatment resulted in reduced levels of P22301 and P05112 , but increased levels of IL-12 , P01579 , P01375 , and inducible NO synthase , reflecting a switch of P01730 (+) differentiation from Th2 to Th1 . This treatment is likely to activate immunity , thereby imparting resistance to reinfection . GRA induced NF-kappaB migration into the nucleus of parasite-infected cells and caused a diminishing presence of IkappaB in the cytoplasm . The lower level of cytoplasmic P25963 in GRA-treated cells resulted from increased phosphorylation of P25963 and higher activity of O15111 ( IKK ) . Additional experiments demonstrated that GRA does not directly affect IKK activity . These results suggest that GRA exerts its effects at some level upstream of IKK in the signaling pathway and induces the production of proinflammatory mediators through a mechanism that , at least in part , involves induction of NF-kappaB activation . DB08895 in kidney transplantation . INTRODUCTION : This review will discuss the mechanism of action and important kidney transplant clinical trial data for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . AREAS COVERED : Successful kidney transplantation requires adequate immunosuppression . Current maintenance immunosuppressive protocols which rely on calcineurin inhibitors have long-term nephrotoxicity and negative impact on cardiometabolic risk factors . JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared to control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . EXPERT OPINION : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . Cantharidin inhibits angiogenesis by suppressing P15692 -induced P23458 / P40763 , P29323 and AKT signaling pathways . Cantharidin ( CTD ) , a chemical compound secreted by blister beetles , has been shown with anti-tumor property in many cancer cells . In this study , our data showed that CTD exerts potent anti-angiogenesis activity in a dose-dependent manner . CTD dose dependently suppressed human umbilical vascular endothelial cells proliferation , migration , and tube formation in vitro . Furthermore , CTD concentration dependently inhibited angiogenesis in chick embryo P62158 model in vivo . At the molecular level , CTD abrogated P15692 -induced activation of P40763 and suppressed the phosphorylation of P23458 and P29323 in a dose-dependent manner . Furthermore , CTD blocked the phosphorylation of AKT in a time-dependent manner . Taken together , these findings clearly demonstrate for the first time that CTD can inhibit angiogenesis and may have applications in the development of new anti-angiogenesis drugs . Kinase inhibitors for the treatment of inflammatory and autoimmune disorders . Drugs targeting inhibition of kinases for the treatment of inflammation and autoimmune disorders have become a major focus in the pharmaceutical and biotech industry . Multiple kinases from different pathways have been the targets of interest in this endeavor . This review describes some of the recent developments in the search for inhibitors of O14920 , Syk , Lck , and P52333 kinases . It is anticipated that some of these compounds or newer inhibitors of these kinases will be approved for the treatment of rheumatoid arthritis , psoriasis , organ transplantation , and other autoimmune diseases . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . A pharmacokinetic and clinical assessment of tofacitinib for the treatment of rheumatoid arthritis . INTRODUCTION : Rheumatoid arthritis ( RA ) is a chronic painful and debilitating autoimmune disease . Although the outcome for patients with RA has improved markedly in the past decades , driven largely by the advent of biological disease-modifying antirheumatic drugs ( DMARDs ) and updated management strategies , adequate disease control can not be achieved in a substantial proportion of patients . Since RA is a syndrome with different biological subsets , DMARDs , with a novel mechanism of action , may represent a valuable addition to the current armamentarium . DB08895 is a novel synthetic DMARD that selectively inhibits Janus kinases ( JAKs ) , particularly P23458 and P52333 . AREAS COVERED : This review describes the pharmacokinetics of tofacitinib . Furthermore , the article summarizes and comments the drug 's efficacy and safety profile in RA patients . The authors furthermore assess data derived from the FDA 's RA development program . EXPERT OPINION : DB08895 is an oral synthetic DMARD displaying linear pharmacokinetics . Metabolism , primarily mediated by P08684 , accounts for 70 % of the total clearance of the drug ; the remaining 30 % are renally excreted . DB08895 monotherapy , or in combination with traditional DMARDs , has demonstrated its efficacy while having an acceptable safety profile in RA patients who have responded inadequately to current DMARDs , including P01375 antagonists . In view of its undetermined benefit to risk ratio , in the real-world population , tofacitinib should , for now , only be prescribed to selected patients . Mobilization of Ph chromosome-negative peripheral blood stem cells in chronic myeloid leukaemia patients with imatinib mesylate-induced complete cytogenetic remission . Imatinib mesylate ( IM , DB00619 , Glivec ) can induce a high rate of complete cytogenetic response ( CCR ) in chronic myeloid leukaemia ( CML ) patients , although to date the majority of patients continue to have detectable disease by sensitive reverse transcription polymerase chain reaction ( RT-PCR ) . It is therefore possible that these patients may ultimately relapse and require treatment such as autologous peripheral blood stem cell transplant ( APBSCT ) . We attempted mobilization of haemopoietic progenitor cells from 58 patients in CCR using recombinant human granulocyte colony-stimulating factor [ rHu- DB00099 ; 10 micro g/kg/d subcutaneously ( s.c. ) for at least 4 d ] alone , while continuing IM treatment . The median d 5 ( peak ) P28906 + count was 11.5/ microl ( range 0-108/ microl ) , and 43/58 ( 74 % ) patients underwent a median of two ( range 1-3 ) apheresis procedures . A median dose of 2.1 x 10(6)/kg P28906 + cells ( range 0.1-6.5 x 10(6)/kg ) was collected . Some 84 % of 31 collections analysed were negative for the Philadelphia ( Ph ) chromosome or breakpoint cluster region and Abelson murine leukaemia viral oncogene homologue ( P11274 - P00519 ) translocation by cytogenetics or fluorescent in situ hybridization respectively . No toxicity was reported with the regimen . Overall , the target P28906 + dose ( 2 x 10(6)/kg P28906 + ) was attained in 23/58 ( 40 % ) patients who entered the study . In summary , we have demonstrated that successful mobilization of Ph- P28906 + cells from IM-treated patients in CCR is possible using rHu-G- P04141 alone . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . Increased levels of Candida albicans mannan-specific T-cell-derived antigen binding molecules in patients with invasive candidiasis . In addition to cytokines , P01730 + T cells have been found to secrete soluble , T-cell-derived antigen binding molecules ( TABMs ) . These antigen-specific immunoproteins are thought to have immunoregulatory properties in the suppression of cell-mediated immunity ( CMI ) because they often associate with interleukin-10 ( P22301 ) and transforming growth factor beta . Decreased CMI causes susceptibility to infections caused by organisms which are normally nonpathogenic . In this situation , e.g. , Candida albicans saprophytism may develop into invasive candidiasis . The difficult diagnosis of invasive candidiasis is based on the findings obtained from blood cultures and with tissue biopsy specimens , with some additional diagnostic value gained by the detection of Candida albicans mannan antigenemia and antimannan antibodies . In the present study , Candida albicans mannan-specific TABM ( P62158 -TABM ) levels in the sera of patients with invasive candidiasis ( n = 11 ) , Candida colonization ( n = 11 ) and noncolonization ( n = 10 ) , recurrent vulvovaginal candidiasis ( n = 30 ) , and atopic eczema dermatitis syndrome ( n = 59 ) and healthy controls ( n = 30 ) were analyzed . For 14 participants , the effect of mannan stimulation on TABM production and gamma interferon ( P01579 ) and P05112 mRNA expression by peripheral blood lymphocytes was also studied . It was demonstrated that P62158 -TABM production was the highest in patients with invasive candidiasis and that P62158 -TABM levels could distinguish Candida-colonized patients from noncolonized patients . In addition , the P62158 -TABM level was directly related to mRNA expression for P05112 but not P01579 . These results reinforce the view that TABMs are associated with decreased CMI , immunoregulation , and the T-helper cell 2-type immune response . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Synergy between P25942 ligation and P05112 on fibroblast proliferation involves P05112 receptor signaling . Fibrosis can be an undesired consequence of activated cellular immune responses . The purpose of this work was to determine whether P25942 ligation and the pro-fibrotic cytokine P05112 interact in regulating fibroblast proliferation and collagen production , and , if so , the mechanisms used . This study found that the combination of P05112 and ligation of P25942 on the fibroblast cell surface had synergistic effects in stimulating fibroblast proliferation . In contrast , P25942 ligation negated the inhibitory effects of P01579 on fibroblast proliferation . Western blotting analyses of fibroblast crude lysates revealed that a potential mechanism of the synergy between P25942 ligation and P05112 was the phosphorylation of proteins at 130 kDa and , to a lesser degree , at 95 , 85 , and 75 kDa . Immunoprecipitation-Western blotting experiments showed that phosphorylation levels of IL-4Ralpha , P23458 , insulin receptor substrate 1 , and insulin receptor substrate 2 , factors with molecular mass close to the observed 130 kDa major phosphorylation band , increased in response to the combined P25942 ligation and P05112 action . In contrast , there was no evidence that synergy was mediated by an increased expression of IL-4Ralpha chain , P25942 , or the autocrine profibrotic cytokines P05231 and TGF-beta . These findings suggest that P25942 - P29965 contacts between fibroblasts and cells secreting P05112 may promote the profibrotic effects of P05112 by affecting signal transduction and reducing the anti-fibrotic effects of P01579 . Tofacitinab in renal transplantation . DB08895 ( tositinib , CP-690,550 ) is a small molecule inhibitor of Janus associated kinases , primarily P52333 and O60674 , which inhibits cytokine signaling through the IL-2Rγ chain . In this article , we review the mechanism of action of tofacitinib , and pre-clinical and clinical data regarding its use in solid organ transplantation thus far . It is hoped that tofacitinib may form the basis for calcineurin-free immunosuppression , improving renal function while eliminating calcineurin inhibitor renal toxicity . Current studies suggest that tofacitinib is an effective immunosuppressive agent for renal transplantation , but it 's use in current protocols carries an increased risk of CMV , BK , and EBV viral infection , anemia and leukopenia , and post-transplant lymphoproliferative disorder . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . Mechanisms of interleukin-1beta-induced P39905 release from rat glioma cells . P39905 ( P39905 ) is highly expressed both in neurons and astrocytes in injured tissues . Astrocytes support neurons by releasing neurotrophic factors including P39905 . It has been reported that various agents including cytokines such as interleukin ( IL ) -1beta induce P39905 mRNA expression and the release in astrocytes . However , the mechanism behind the P39905 synthesis and release remains unclear . Herein , we investigated the mechanisms of the IL-1beta-induced P39905 release from rat P13671 glioma cells . IL-1beta time dependently stimulated P39905 release from P13671 cells . IL-1beta induced the phosphorylation of inhibitor kappa B ( IkappaB ) , p38 mitogen-activated protein ( Q96HU1 ) kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase , stress-activated protein kinase/c-Jun N-terminal kinase ( SAPK/JNK ) and signal transducer and activator of transcription ( P35610 ) 3 . The IL-1beta-stimulated levels of P39905 were suppressed by wedelolactone , an inhibitor of O15111 , SB203580 , an inhibitor of p38 Q96HU1 kinase , PD98059 , an inhibitor of Q02750 /2 or Janus family of tyrosine kinase ( JAK ) inhibitor I , an inhibitor of upstream kinase of P40763 . On the contrary , SP600125 , an inhibitor of SAPK/JNK , failed to reduce the IL-1beta-effect . These results strongly suggest that IL-1beta stimulates P39905 release through the pathways of IkappaB-nuclear factor kappa B , p38 Q96HU1 kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase and JAK- P40763 , but not through the SAPK/JNK pathway in glioma cells . Efficacy and safety of tofacitinib for treatment of rheumatoid arthritis . DB08895 is the first in a new class of nonbiologic disease-modifying antirheumatic drugs ( DMARDs ) , a targeted , synthetic DMARD , approved for the treatment of rheumatoid arthritis ( RA ) as monotherapy or in combination with methotrexate or other non-biologic DMARD . DB08895 , an orally administered Janus kinase ( JAK ) inhibitor , decreases T-cell activation , pro-inflammatory cytokine production , and cytokine signaling by inhibiting binding of type I cytokine receptors family and γ-chain cytokines to paired P23458 / P52333 receptors . The net effect of tofacitinb 's mechanism of action is decreased synovial inflammation and structural joint damage in RA patients . To date , six phase 3 trials have been conducted to evaluate the safety and efficacy of tofacitinib under the oral rheumatoid arthritis triaLs ( ORAL ) series . This review describes the pharmacology of the novel agent , tofacitinib , and details the safety and efficacy data of the ORAL trials . Constitutively activated P40763 frequently coexpresses with epidermal growth factor receptor in high-grade gliomas and targeting P40763 sensitizes them to DB00317 and alkylators . PURPOSE : The goals of this study are to elucidate the relationship of the oncogenic transcription factor signal transducer and activator of transcription 3 ( P40763 ) with glioma aggressiveness and to understand the role of high P40763 activity in the resistance of malignant gliomas and medulloblastomas to chemotherapy . EXPERIMENTAL DESIGN : Immunohistochemical staining and biochemical methods were used to examine the extent of P40763 activation and P00533 expression in primary specimens and cell lines , respectively . Cellular response to drug treatments was determined using cell cytotoxicity and clonogenic growth assays . RESULTS : We found P40763 to be constitutively activated in 60 % of primary high-grade/malignant gliomas and the extent of activation correlated positively with glioma grade . High levels of activated/phosphorylated P40763 were also present in cultured human malignant glioma and medulloblastoma cells . Three P40763 -activating kinases , Janus-activated kinase 2 ( O60674 ) , P00533 , and EGFRvIII , contributed to P40763 activation . An inhibitor to O60674 / P40763 , JSI-124 , significantly reduced expression of P40763 target genes , suppressed cancer cell growth , and induced apoptosis . Furthermore , we found that P40763 constitutive activation coexisted with P00533 expression in 27.2 % of primary high-grade/malignant gliomas and such coexpression correlated positively with glioma grade . Combination of an anti- P00533 agent DB00317 and a O60674 / P40763 inhibitor synergistically suppressed P40763 activation and potently killed glioblastoma cell lines that expressed P00533 or EGFRvIII . JSI-124 also sensitized malignant glioma and medulloblastoma cells to temozolomide , 1,3-bis(2-chloroethyl)-1-nitrosourea , and cisplatin in which a synergism existed between JSI-124 and cisplatin . CONCLUSION : P40763 constitutive activation , alone and in concurrence with P00533 expression , plays an important role in high-grade/malignant gliomas and targeting P40763 / O60674 sensitizes these tumors to anti- P00533 and alkylating agents . Selective JAK/ P40763 signalling regulates transcription of colony stimulating factor-2 and -3 in Concanavalin-A-activated mesenchymal stromal cells . Human bone marrow-derived mesenchymal stromal cells ( MSCs ) express Toll-like receptors ( TLRs ) and produce cytokines and chemokines , all of which contribute to these cells ' immunomodulatory and proangiogenic properties . Among the secreted cytokines , colony-stimulating factors ( CSFs ) regulate angiogenesis through activation of endothelial cell proliferation and migration . Since O60682 are recruited within hypoxic tumors where they signal paracrine-regulated angiogenesis , the aim of this study was to evaluate which P04141 members are expressed and are inducible in activated O60682 . Furthermore , we investigated the JAK/ P35610 signal transducing pathway that may impact on P04141 transcription . O60682 were activated with Concanavalin-A ( ConA ) , a TLR-2/6 agonist as well as a membrane type-1 matrix metalloproteinase ( P50281 ) inducer , and we found increased transcription of granulocyte macrophage- P04141 ( GM- P04141 , P04141 -2 ) , granulocyte P04141 ( DB00099 , P04141 -3 ) , and P50281 . Gene silencing of either P40763 or P50281 prevented ConA-induced phosphorylation of P40763 , and reversed ConA effects on P04141 -2 and P04141 -3 . Treatment with the Janus Kinase (JAK)2 inhibitor AG490 antagonized the ConA induction of P50281 and P04141 -2 , while the pan-JAK inhibitor DB08895 reversed ConA-induced P04141 -2 and -3 gene expression . Silencing of O60674 prevented the ConA-mediated increase of P04141 -2 , while silencing of P23458 , P52333 and P29597 prevented the increase in P04141 -3 . Given that combined TLR-activation and locally-produced P04141 -2 and P04141 -3 could regulate immunomodulation and neovascularization , pharmacological targeting of TLR-2/6-induced P50281 /JAK/ P40763 signalling pathway may prevent O60682 contribution to tumor development . DB00317 ( DB00317 ) inhibits phosphorylation of three different downstream signal transducers in head and neck cancer ( SCCHN ) . Proliferation of squamous cell carcinoma of the head and neck ( SCCHN ) depends on epidermal growth factor receptor ( P00533 ) expression . The P00533 activates different signal pathways leading to gene transcription in the nucleus due to cell cycle progression and proliferation of tumor cells . AKT , P40763 and MAPK play central roles in these pathways . However , they are not only regulated by the P00533 . We therefore investigated whether a specific inhibitor of the P00533 tyrosine kinase ( DB00317 or DB00317 ) is able to inhibit phosphorylation of these three signals at the same time . Western blot analysis of pretreated SCCHN cells revealed that DB00317 greatly reduces the amount of phosphorylated AKT , P40763 as well as MAPK Surprisingly , this effect was not dose-dependent between the concentration range of 5.15 to 41.2 microM/ml . We conclude that DB00317 has a high potency to inhibit nuclear gene transcription responsible for cell cycle progression . Furthermore , dose reduction of DB00317 in the case of toxicity may not severely influence the response to treatment . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Expression of the adaptor protein Lnk in leukemia cells . OBJECTIVE : Tyrosine kinases are involved in cytokine signaling and are frequently aberrantly activated in hematological malignancies . Lnk , a negative regulator of cytokine signaling , plays critical nonredundant roles in hematopoiesis . By binding to phosphorylated tyrosine kinases , Lnk inhibits major cytokine receptor signaling , including c- P10721 ; erythropoietin receptor- O60674 ( O60674 ) ; and P40238 - O60674 . In the present study , we investigated Lnk expression and possible function in transformed hematopoietic cells . MATERIALS AND METHODS : Coimmunoprecipitations were performed to identify binding between Lnk and mutant tyrosine kinases . Proliferation assays were done to examine the affect of Lnk overexpression on cancer cell growth . Real-time polymerase chain reaction analysis was used to determine Lnk expression in patient samples . RESULTS : We show that , in parallel to binding wild-type O60674 and c- P10721 , Lnk associates with and is phosphorylated by mutant alleles of O60674 and c- P10721 . In contrast , Lnk does not bind to and is not phosphorylated by P11274 - P00519 fusion protein . Ectopic expression of Lnk strongly attenuates growth of some leukemia cell lines , while others as well as most solid tumor cancer cell lines are either moderately inhibited or completely insensitive to Lnk . Furthermore , Lnk-mediated growth inhibition is associated with differential downregulation of phosphatidylinositol 3 kinase/Akt/mammalian target of rapamycin and mitogen-activated protein kinase/extracellular signal-regulated kinase signaling in leukemia cell lines . Surprisingly , analysis of Lnk in a large panel of myelodysplastic syndrome and acute myeloid leukemia patient samples revealed high levels of Lnk in nearly half of the samples . CONCLUSION : Although how leukemic cells overcome the antiproliferative effects of Lnk is not yet clear , our data highlight the multifaceted role negative feedback mechanisms play in malignant transformation . Effects of serotonin and serotonergic agonists and antagonists on the production of interferon-gamma and interleukin-10 . Serotonin ( 5-HT ) is a neurotransmitter and an immune modulator . In vitro , antidepressants with a serotonergic mode of action have , at concentrations within the therapeutical range , negative immunoregulatory effects , i.e. , they increase the production rate of interleukin-10 ( P22301 ) , a negative immunoregulatory cytokine . We have hypothesized that part of these effects may be explained by the serotonergic activities of antidepressants on immunocytes . This study was carried out to examine the effects of 5-HT , p-chlorophenylalanine ( PCPA ) , a 5-HT depleting agent , flesinoxan ( a P08908 agonist ) , m-chlorophenylpiperazine ( mCPP ; a 5- Q13049 /2C agonist ) , and ritanserin ( a 5- Q13049 /2C antagonist ) on the production rate of interferon-gamma ( IFNgamma ) , a proinflammatory cytokine , and P22301 by whole blood stimulated with polyclonal activators . The IFNgamma/ P22301 production ratio was computed , since this ratio reflects the pro- versus anti-inflammatory capacity of cultured whole blood . We found that : 1 ) 5-HT , 150 ng/mL , 1.5 microg/mL , and 15 microg/mL significantly decreased the IFNgamma/ P22301 ratio ; 2 ) PCPA ( 5 microM ) significantly suppressed the production of IFNgamma and P22301 ; 3 ) flesinoxan ( 15 ng/mL ; 1.5 microg/mL ) had no significant effects on the production of the above cytokines ; and 4 ) mCPP ( 2.7 microg/mL ) and ritanserin ( 5.0 microg/mL ) suppressed the IFNgamma/ P22301 ratio . It is concluded that intracellular 5-HT may be necessary for an optimal synthesis of IFNgamma and P22301 , and that extracellular 5-HT concentrations at or above serum values may suppress the production of the proinflammatory cytokine IFNgamma . The negative immunoregulatory effects of antidepressive drugs are probably not related to their serotonergic activities . Identification of a viability domain in the granulocyte/macrophage colony-stimulating factor receptor beta-chain involving tyrosine-750 . The granulocyte/macrophage colony-stimulating factor ( GM- P04141 ) receptor ( GMR ) is a heterodimeric receptor expressed by myeloid lineage cells . In this study we have investigated domains of the GMR beta-chain ( GMR beta ) involved in maintaining cellular viability . Using a series of nested GMR beta deletion mutants , we demonstrate that there are at least two domains of GMR beta that contribute to viability signals . Deletion of amino acid residues 626-763 causes a viability defect that can be rescued with fetal calf serum ( FCS ) . Deletion of residues 518-626 , in contrast , causes a further decrement in viability that can be only partially compensated by the addition of FCS . GMR beta truncated proximal to amino acid 517 will not support long-term growth under any conditions . Site-directed mutagenesis of tyrosine-750 ( Y750 ) , which is contained within the distal viability domain , to phenylalanine eliminates all demonstrable tyrosine phosphorylation of GMR beta . Cell lines transfected with mutant GMR beta ( Y750 --> F ) have a viability disadvantage when compared to cell lines containing wild-type GMR that is partially rescued by the addition of FCS . We studied signal transduction in mutant cell lines in an effort to identify pathways that might participate in the viability signal . Although tyrosine phosphorylation of O60674 , Q06124 , and Vav is intact in Y750 --> F mutant cell lines , Shc tyrosine phosphorylation is reduced . This suggests a potential role for Y750 and potentially Shc in a GM- P04141 -induced signaling pathway that helps maintain cellular viability . Modulation of cytokine release from human monocytes by drugs used in the therapy of inflammatory bowel diseases . BACKGROUND : Cytokines produced in the gut mucosa play an important role in the pathogenesis of inflammatory bowel diseases ( Q9UKU7 ) . To determine whether drugs used in the treatment of these diseases modulate cytokine synthesis , we investigated their effects on endotoxin-induced tumour necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -1 beta and P05231 release by elutriation-purified human monocytes in vitro . METHODS : Drugs tested were dexamethasone , DB00244 , sulphapyridine and zileuton ( a P09917 inhibitor ) . Monocytes were isolated and stimulated with endotoxin , and P01375 , IL-1 and P05231 levels were determined using an enzyme-linked immunosorbent assay . RESULTS : Monocyte stimulation with endotoxin resulted in an average P01375 release of 2464 +/- 64 pg/10(6) cells , IL-1 release of 616 +/- 47 pg/10(6) cells and P05231 release of 2259 +/- 148 pg/10(6) cells . Addition of dexamethasone resulted in a reduction of P01375 , IL-1 and P05231 release to below background levels . DB00891 significantly reduced P01375 and induced IL-1 release in a dose-dependent fashion , but had no significant effect on P05231 release . 5- DB00233 did not modulate P05231 synthesis , but significantly reduced IL-1 and enhanced P01375 synthesis . Zileuton reduced P01375 and P05231 release , but enhanced IL-1 release . CONCLUSION : We conclude that these anti-inflammatory drugs are able to modulate cytokine release by human monocytes . Further studies are needed to determine whether these effects are related to their therapeutic efficacy in Q9UKU7 . P52333 inhibition significantly attenuates psoriasiform skin inflammation in P05107 mutant PL/J mice . P52333 , a member of the Janus kinase family , is predominantly expressed in hemopoietic cells and binds specifically to the common gamma chain of a subfamily of cytokine receptors that includes P60568 , P05112 , P13232 , P15248 , P40933 , and Q9HBE4 . Previous studies suggest that this tyrosine kinase plays key roles in mediating T cell functions , and inhibition of P52333 has been shown to prevent graft rejection and decrease the severity of arthritis in rodent models . However , the functions of P52333 in the development of skin immune responses and diseases such as psoriasis have not been determined . P05107 mutant PL/J mice develop spontaneous T cell-dependent psoriasiform skin disease with several similarities to human psoriasis . In this study , we treated mice with established skin disease with R348 , a small molecule inhibitor of P52333 , and observed a marked attenuation of skin lesions following 6 wk of treatment . Histological analyses revealed major reductions of both epidermal and dermal lesion severity scores in R348-treated P05107 -deficient PL/J mice compared with vehicle controls , which was associated with decreased P01730 (+) T cell infiltration . In addition , systemic levels of Q16552 , Q9GZX6 , IL-23 , and P01375 were significantly lower in mice receiving the compound , and T cells isolated from R348-treated mice also showed reduced phosphorylation of Stat5 after stimulation with P60568 . These findings suggest that small-molecule inhibitors of P52333 may be useful in the treatment of inflammatory skin diseases such as psoriasis and strongly implicate JAK signaling events as important in the pathogenesis of this disease . Pyrrolo[1,2-f]triazines as O60674 inhibitors : achieving potency and selectivity for O60674 over P52333 . SAR studies of pyrrolo[1,2-f]triazines as O60674 inhibitors is presented . Achieving O60674 inhibition selectively over P52333 is discussed . DB08895 . DB08895 ( CP-690,550 ; CP-690550 ; CP690550 ) , an orally active immunosuppressant , is being developed by Pfizer for the treatment of rheumatoid arthritis , inflammatory bowel disease , dry eyes , ankylosing spondylitis , psoriasis , psoriatic arthritis , and for the prevention of transplant rejection . DB08895 specifically inhibits Janus activated kinase 3 ( P52333 ) , which has a pivotal role in cytokine signal transduction that governs lymphocyte survival , proliferation , differentiation , and apoptosis . This review discusses the key development milestones and therapeutic trials of this drug . Preclinical to clinical translation of tofacitinib , a Janus kinase inhibitor , in rheumatoid arthritis . A critical piece in the translation of preclinical studies to clinical trials is the determination of dosing regimens that allow maximum therapeutic benefit with minimum toxicity . The preclinical pharmacokinetic ( PK ) /pharmacodynamic ( PD ) profile of tofacitinib , an oral Janus kinase ( JAK ) inhibitor , in a mouse collagen-induced arthritis ( mCIA ) model was compared with clinical PK/PD data from patients with rheumatoid arthritis ( RA ) . Preclinical evaluations included target modulation and PK/PD modeling based on continuous subcutaneous infusion or oral once- or twice-daily ( P55957 ) dosing paradigms in mice . The human PK/PD profile was obtained from pooled data from four phase 2 studies in patients with RA , and maximal effect models were used to evaluate efficacy after 12 weeks of tofacitinib treatment ( 1-15 mg P55957 ) . In mCIA , the main driver of efficacy was inhibition of cytokine receptor signaling mediated by P23458 heterodimers , but not O60674 homodimers , and continuous daily inhibition was not required to maintain efficacy . Projected efficacy could be predicted from total daily exposure irrespective of the oral dosing paradigm , with a total steady-state plasma concentration achieving 50 % of the maximal response ( Cave50 ) of ~100 nM . DB08895 potency ( ED50 ) in clinical studies was ~3.5 mg P55957 ( 90 % confidence interval : 2.3 , 5.5 ) or total Cave50 of ~40 nM , derived using Disease Activity Scores from patients with RA . The collective clinical and preclinical data indicated the importance of Cave as a driver of efficacy , rather than maximum or minimum plasma concentration ( Cmax or Cmin ) , where Cave50 values were within ~2-fold of each other . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . Dysfunction of the P19957 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia . Patients with myeloproliferative disorders ( MPDs ) , such as essential thrombocythemia ( ET ) have increased risk of thrombosis and bleeding , which are major sources of morbidity and mortality . Most P53602 patients have a gain of function mutation in O60674 ( JAK2V617F ) , but little is known how JAK2V617F affects platelet function . Here , we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin ( which couples to O60674 ) on this pathway . In contrast , SFLLRN-mediated P16109 expression , DB00171 secretion , phosphorylation of the PKC substrate pleckstrin , and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets . In addition , thrombopoietin-mediated O60674 phosphorylation was unchanged , suggesting that signaling pathways activated downstream of O60674 are impaired . Indeed , we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the P19957 kinase substrate Akt , and have reduced activation of Rap1 in response to thrombopoietin , DB01277 ,ADP , SFLLRN , and thrombin . This effect was independent of Giα Q9H244 purinergic receptor function as ADP-mediated inhibition of P50552 phosphorylation was unchanged . These results demonstrate that the P19957 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients , resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation . The JAK inhibitor , tofacitinib , reduces the T cell stimulatory capacity of human monocyte-derived dendritic cells . OBJECTIVE : DB08895 , which is a Janus kinase ( JAK ) inhibitor , has shown clinical effects in the treatment of rheumatoid arthritis . JAKs are important kinases in lymphocyte differentiation ; however , their function in dendritic cells ( DCs ) is unknown . In this study , the function of JAKs in DCs was investigated with tofacitinib . METHODS : The effects of tofacitinib on the maturation of human monocyte-derived DCs induced by lipopolysaccharide ( LPS ) stimulation were investigated . In addition , its effects on T cell stimulatory capability was investigated by coculturing with naïve CD45RA-positive T cells . RESULTS : DB08895 decreased expression of P33681 / P42081 in a concentration-dependent manner in LPS-stimulated DCs ; however , it did not affect HLA-DR expression . DB08895 suppressed tumour necrosis factor , interleukin ( IL ) -6 and IL-1β production without affecting transforming growth factor ( TGF ) -β and P22301 production . Meanwhile , P33681 / P42081 expression in DCs was enhanced by type I interferon ( IFN ) stimulation , and the LPS-induced P33681 / P42081 expression was inhibited by an antibody to type I IFN receptor . Furthermore , tofacitinib suppressed production of type I IFN and activation of interferon regulatory factor ( Q969Q1 ) -7 , which is a transcription factor involved in P33681 / P42081 and type I IFN expression . DB08895 also decreased the T cell stimulatory capability of DCs and increased expression of indoleamine 2,3-dioxygenase ( P14902 ) -1 and Q6ZQW0 . CONCLUSIONS : DB08895 , a P23458 / P52333 inhibitor , affected the activities of human DCs . It decreased P33681 / P42081 expression and T cell stimulatory capability through suppression of type I IFN signalling . These results suggest a novel mode of action for tofacitinib and a pivotal role for JAKs in the differentiation of DCs . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Novel path to P05231 trans-signaling through thrombin-induced soluble P05231 receptor release by platelets . Interleukin ( IL ) -6 is a multifunctional cytokine with a critical role in inflammatory , immunoregulatory and haemopoietic responses . Its receptor consists of an ubiquitously expressed membrane transducing element ( P40189 ) and of the specific element P08887 ( P08887 ) , present only on hepatocytes and some leukocyte subsets . P08887 also exists as soluble protein ( sIL-6R ) that , in the presence of P05231 , forms a complex able to bind P40189 and , thanks to the mechanism called trans-signaling , transduces P05231 effect through tyrosine phosphorylation and activation of the signal transducer and transcription activator ( P35610 ) -3 . The aim of this study was to analyze the bidirectional relationships between platelet aggregation and P05231 -dependent effects . While platelets do not produce P05231 , we found that resting platelets express P40189 , but not P08887 , on their membranes . Upon activation by thrombin or calcium ionophore A23187 , but not by ADP , the P08887 is released in soluble form , while cangrelor , the specific inhibitor of Q9H244 receptor , can partially inhibit sIL-6R release . This sIL-6R is biologically active and , in the presence of P05231 , can trigger P05231 trans-signaling , inducing an autocrine activation loop ( as measured by an increase in P08887 and P40189 content ) and P40763 phosphorylation . On the other hand , P05231 trans-signaling has no effect on platelet degranulation or aggregation by itself , nor on thrombin-induced platelet aggregation . Our data add an important piece to the puzzle of thrombosis and inflammation : in the presence of P05231 , which can be produced by stressed endothelial cells , the platelet-derived P05231 trans-signaling could be crucial for the evolution of inflammation within a damaged vessel . P04141 activates NF-kappaB via direct interaction of the P04141 receptor with O15111 beta . P04141 ( P04141 ) has a central role in proliferation and differentiation of hematopoetic cells . Furthermore , it influences the proliferation and migration of endothelial cells . P04141 elicits these functions by activating a receptor consisting of a ligand-specific alpha-chain and a beta-chain , which is common for P04141 , interleukin-3 ( P08700 ) , and P05113 . It is known that various signaling molecules such as O60674 or transcription factors of the signal transducer and activator of transcription ( P35610 ) family bind to the common beta-chain and initiate signaling cascades . However , alpha-chain-specific signal transduction adapters have to be postulated given that P08700 , P05113 , and P04141 induce partly distinct biologic responses . Using a yeast 2-hybrid system , we identified the alpha-chain of the P04141 receptor ( GMRalpha ) as putative interaction partner of O15111 beta , one of the central signaling kinases activating the transcription factor nuclear factor-kappaB ( NF-kappaB ) . Using endogenous protein levels of endothelial cell extracts , we could verify the interaction by coimmunoprecipitation experiments . Fluorescence resonance energy transfer ( FRET ) microscopy confirmed the direct interaction of P27918 -IKKbeta and YFPGMRalpha in living cells . Functional studies demonstrated P04141 -dependent activation of O15111 activity in endothelial cells , degradation of IkappaB , and activation of NF-kappaB . Further biologic studies using P04141 -dependent TF-1 cells indicated that P04141 -triggered activation of NF-kappaB is important for cell survival and proliferation . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Curcumin triggers p16-dependent senescence in active breast cancer-associated fibroblasts and suppresses their paracrine procarcinogenic effects . Activated cancer-associated fibroblasts ( CAFs ) or myofibroblasts not only facilitate tumor growth and spread but also affect tumor response to therapeutic agents . Therefore , it became clear that efficient therapeutic regimens should also take into account the presence of these supportive cells and inhibit their paracrine effects . To this end , we tested the effect of low concentrations of curcumin , a pharmacologically safe natural product , on patient-derived primary breast Q13111 cells . We have shown that curcumin treatment upregulates p16(INK4A) and other tumor suppressor proteins while inactivates the O60674 / P40763 pathway . This reduced the level of alpha-smooth muscle actin ( α-SMA ) and the migration/invasion abilities of these cells . Furthermore , curcumin suppressed the expression/secretion of stromal cell-derived factor-1 ( P48061 ) , interleukin-6 ( P05231 ) , matrix metalloproteinase-2 ( P08253 ) , P14780 , and transforming growth factor-β , which impeded their paracrine procarcinogenic potential . Intriguingly , these effects were sustained even after curcumin withdrawal and cell splitting . Therefore , using different markers of senescence [ senescence-associated β-galactosidase ( SA-β-gal ) activity , Ki-67 and Lamin B1 levels , and bromodeoxyuridine incorporation ] , we have shown that curcumin markedly suppresses Lamin B1 and triggers DNA damage-independent senescence in proliferating but not quiescent breast stromal fibroblasts . Importantly , this curcumin-related senescence was p16(INK4A)-dependent and occurred with no associated inflammatory secretory phenotype . These results indicate the possible inactivation of cancer-associated myofibroblasts and present the first indication that curcumin can trigger DNA damage-independent and safe senescence in stromal fibroblasts . JAK inhibitors : treatment efficacy and safety profile in patients with psoriasis . Janus kinase ( JAK ) pathways are key mediators in the immunopathogenesis of psoriasis . Psoriasis treatment has evolved with the advent of targeted therapies , which inhibit specific components of the psoriasis proinflammatory cascade . JAK inhibitors have been studied in early phase trials for psoriasis patients , and the data are promising for these agents as potential treatment options . DB08895 , an oral or topically administered P23458 and P52333 inhibitor , and ruxolitinib , a topical P23458 and O60674 inhibitor , have been most extensively studied in psoriasis , and both improved clinical symptoms of psoriasis . Additional P23458 or P52333 inhibitors are being studied in clinical trials . In phase III trials for rheumatoid arthritis , tofacitinib was efficacious in patients with inadequate responses to tumor necrosis factor inhibitors , methotrexate monotherapy , or disease-modifying antirheumatic drugs . The results of phase III trials are pending for these therapies in psoriasis , and these agents may represent important alternatives for patients with inadequate responses to currently available agents . Further investigations with long-term clinical trials are necessary to verify their utility in psoriasis treatment and assess their safety in this patient population . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Trisomy 6 in Merkel cell carcinoma : a recurrent chromosomal aberration . We retrospectively investigated 17 cases of primary and metastasizing Merkel cell carcinomas ( MCC ) from 14 patients using chromosomal in-situ hybridization ( Q9NSE2 ) to study the occurrence of trisomy 6 in these lesions . METHODS AND RESULTS : Histological diagnosis on all tumour samples was obtained on haematoxylin and eosin stained sections . Immunohistochemistry was performed with antibodies against pancytokeratin ( P62158 5.2 ) , cytokeratin 20 ( CK20 ) , P14209 antigen ( P14209 ) , neuron-specific enolase ( P09104 ) , and chromogranin A ( chrA ) . Sections ( 4 microm ) of the paraffin-embedded tumours were analysed with alpha-satellite centromeric probes for chromosome 6 or 17 using Q9NSE2 . The signal was amplified by the Tyramide Signal Amplification ( P32119 ) assay . Immunohistochemically , the tumours showed the same general epithelial neuro-endocrine pattern : 11/13 expressed cytokeratin 20 , and 47 % exhibited trisomy 6 , with no significant difference between primary and metastatic lesions . Incomplete follow-up data did not allow us to establish a prognostic value of trisomy 6 , however , this aberration might be an additional diagnostic tool in distinguishing MCC from other small round blue cell tumours . CONCLUSIONS : Q9NSE2 seems to be a promising adjunctive method to diagnose Merkel cell carcinoma . Trisomy 6 should be investigated more closely in these cases , as has been done for chromosomes 1 and 11 . Of particular interest would be identification of modifications in proto-oncogene(s) located on chromosome 6 . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . DB00619 inhibition effect on KITAsn822Lys-mediated signal transduction cascade . OBJECTIVE : Alterations in growth factor signaling pathways may be a frequent collaborating event in Q01196 - Q06455 -mediated leukemogenesis . Gain-of-function P10721 receptor mutations have been reported in adult AML patients , especially those with core binding factor leukemia ( CBFL ) . We have previously reported a new gain-of-function P10721 (Asn822Lys) mutation that is constitutively expressed in the Kasumi-1 CBFL cell line , and has recently been described in two childhood AML patients . To explore the molecular basis of the effects of this mutation in the appropriate context of hemopoietic dysregulation , we investigated P10721 downstream signaling in the Kasumi-1 cell line by means of DB00619 ( Imatinib , Gleevec ) pharmacological inhibition . MATERIALS AND METHODS : We investigated P10721 (Asn822Lys) mutant-initiated signaling in Kasumi-1 cell line , and characterized the inhibitory effect of the DB00619 protein tyrosine kinase inhibitor on downstream signaling . RESULTS : The use of DB00619 -mediated inhibition impaired the tyrosine phosphorylation of P10721 (Asn822Lys) and its association with the p85 subunit of phosphatidylinositol 3'-kinase ( p85PI3K ) . The downstream constitutive phosphorylation of P45983 /2 and P40763 was also significantly inhibited , but DB00619 had no effect on the constitutive activation of Akt , thus suggesting that it is due to other signaling in Kasumi-1 cells . DB00619 inhibited the P10721 -mediated proliferation of Kasumi-1 cells in a dose-dependent manner . CONCLUSIONS : These findings show the role of PI3K in P10721 (Asn822Lys)-mediated constitutive activation through the Akt-independent downstream signaling pathway of JNK , and also demonstrate the mutant 's susceptibility to DB00619 , which may therefore have therapeutic potential in CBFL patients with susceptible P10721 mutations . EGCG enhances the therapeutic potential of gemcitabine and CP690550 by inhibiting P40763 signaling pathway in human pancreatic cancer . BACKGROUND : Signal Transducer and Activator of Transcription 3 ( P40763 ) is an oncogene , which promotes cell survival , proliferation , motility and progression in cancer cells . Targeting P40763 signaling may lead to the development of novel therapeutic approaches for human cancers . Here , we examined the effects of epigallocathechin gallate ( EGCG ) on P40763 signaling in pancreatic cancer cells , and assessed the therapeutic potential of EGCG with gemcitabine or P52333 inhibitor CP690550 ( DB08895 ) for the treatment and/or prevention of pancreatic cancer . METHODOLOGY/PRINCIPAL FINDINGS : Cell viability and apoptosis were measured by XTT assay and TUNEL staining , respectively . Gene and protein expressions were measured by qRT-PCR and Western blot analysis , respectively . The results revealed that EGCG inhibited the expression of phospho and total P52333 and P40763 , P40763 transcription and activation , and the expression of P40763 -regulated genes , resulting in the inhibition of cell motility , migration and invasion , and the induction of caspase-3 and PARP cleavage . The inhibition of P40763 enhanced the inhibitory effects of EGCG on cell motility and viability . Additionally , gemcitabine and CP690550 alone inhibited P40763 target genes and synergized with EGCG to inhibit cell viability and induce apoptosis in pancreatic cancer cells . CONCLUSIONS/SIGNIFICANCE : Overall , these results suggest that EGCG suppresses the growth , invasion and migration of pancreatic cancer cells , and induces apoptosis by interfering with the P40763 signaling pathway . Moreover , EGCG further enhanced the therapeutic potential of gemcitabine and CP690550 against pancreatic cancer . Targeting P52333 in kidney transplantation : current status and future options . PURPOSE OF REVIEW : This review will discuss the mechanism of action and important clinical trial data in renal transplantation for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . RECENT FINDINGS : JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared with vehicle control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new-onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . SUMMARY : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Kinase inhibitors : a new class of antirheumatic drugs . The outlook for patients with rheumatoid arthritis has improved significantly over the last three decades with the use of disease-modifying antirheumatic drugs . However , despite the use of methotrexate , cytokine inhibitors , and molecules targeting T and B cells , a percentage of patients do not respond or lose their response over time . The autoimmune process in rheumatoid arthritis depends on activation of immune cells , which utilize intracellular kinases to respond to external stimuli such as cytokines , immune complexes , and antigens . In the past decade , small molecules targeting several kinases , such as p38 MAPK , Syk , and JAK have been developed . Several p38 MAPK inhibitors proved ineffective in treating rheumatoid arthritis . The Syk inhibitor , fostamatinib , proved superior to placebo in Phase II trials and is currently under Phase III investigation . DB08895 , a P23458 /3 inhibitor , was shown to be efficacious in two Phase III trials , while VX-509 , a P52333 inhibitor , showed promising results in a Phase II trial . Fostamatinib and tofacitinib were associated with increased rates of infection , elevation of liver enzymes , and neutropenia . Moreover , fostamatinib caused elevations of blood pressure and diarrhea , while tofacitinib was associated with an increase in creatinine and elevation of lipid levels . Activation of the JAK/ P35610 pathway in vascular smooth muscle by serotonin . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a vasoconstrictor and mitogen whose levels are elevated in diabetes . Previous studies have shown the presence of 5- Q13049 , P41595 , and P28222 receptors in vascular smooth muscle cells ( VSMCs ) . There are currently no data regarding P41595 and P28222 receptor activation of the JAK/ P35610 pathway in VSMCs and resultant potential alterations in 5-HT signaling in diabetes . Therefore , we tested the hypothesis that 5-HT differentially activates the JAK/ P35610 pathway in VSMCs under conditions of normal ( 5 mM ) and high ( 25 mM ) glucose . Treatment of rat VSMCs with 5-HT ( 10(-6) M ) resulted in time-dependent activation ( approximately 2-fold ) of O60674 , P23458 , and P42224 , but not P40763 ( maximal at 5 min , returned to baseline by 30 min ) . The P41595 receptor agonist BW723C86 and the P28222 receptor agonist CGS12066A ( 10(-9)-10(-5) M , 5-min stimulation ) did not activate the JAK/ P35610 pathway . Treatment with the 5- Q13049 receptor antagonist ketanserin ( 10 nM ) inhibited O60674 activation by 5-HT . Treatment of streptozotocin-induced diabetic rats with ketanserin ( 5 mg.kg-1.day-1 ) reduced activation of O60674 and P42224 but not P40763 in endothelium-denuded thoracic aorta in vivo . 5-HT ( 10(-6) M ) treatment resulted in increased cell proliferation and increased DNA synthesis , which were inhibited by the O60674 inhibitor AG490 . Further studies with apocynin , diphenyleneiodonium chloride , catalase , and virally transfected superoxide dismutase had no effect at either glucose concentration on activation of the JAK/ P35610 pathway by 5-HT . Therefore , we conclude that 5-HT activates O60674 , P23458 , and P42224 via the 5- Q13049 receptors in a reactive oxygen species-independent manner under both normal and high glucose conditions . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process .

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Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Clinical effects and outcomes with new Q9H244 inhibitors in ACS . Thienopyridines have become the cornerstone of treatment for percutaneous coronary intervention although no survival benefit has ever been shown with clopidogrel despite increasing loading doses . Newly developed Q9H244 inhibitors are more potent , more predictable , and have a faster onset of action than clopidogrel , characteristics that make them particularly attractive for high-risk percutaneous coronary intervention ( P05154 ) . Four new Q9H244 inhibitors have been tested each of them having particular individual properties . Prasugrel is an oral pro-drug leading to irreversible blockade of the Q9H244 receptor and is approved worldwide for ACS P05154 . DB08816 is a direct-acting and reversible inhibitor of the Q9H244 receptor with potentially more pleiotropic effects . DB06441 is an intravenous direct and reversible inhibitor of the Q9H244 receptor providing the highest level of inhibition , and elinogrel is an intravenous and oral Q9H244 antagonist with a direct and reversible action . Both prasugrel and ticagrelor , opposed to clopidogrel , have shown that stronger Q9H244 inhibition led respectively to significant 19 and 16 % relative risk reduction of a similar primary end point combining cardiovascular death , non-fatal myocardial infarction , or non-fatal stroke . Both drugs showed a significant 0.6 % absolute excess of TIMI major bleeding not related to CABG surgery . Because in clinical trials , patients perceived to be at higher risk of bleeding usually are excluded , the risk of major and even fatal bleeding might even be higher in a ' real-world ' setting , i.e. in the elderly patient with comorbidities . On the other hand , these newly developed Q9H244 inhibitors decrease mortality after P05154 compared with clopidogrel . The risk/benefit ratio is particularly favorable in P05154 for patients with STEMI . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Inactivation of plasminogen activator inhibitor-1 accelerates thrombolysis of a platelet-rich thrombus in rat mesenteric arterioles . To investigate the role of active plasminogen activator inhibitor 1 ( P05121 ) in the evolution of a microthrombus generated in the arteriolar microcirculation , the monoclonal antibody , 33H1F7 , which transforms active P05121 to a tissue type plasminogen activator ( t-PA ) substrate , was evaluated in an arteriolar thrombosis model in the rat mesentery . Arterioles ( 200-300 um ) were stimulated electrically to create an endothelial lesion ; ADP was then perfused for 2 min to induce the formation of a platelet-rich thrombus which lysed spontaneously in 140 +/- 24 s . Two successive ADP superfusions produced comparable thrombi which lysed in comparable times . Different doses of 33H1F7 were infused to rats for 30 min and the dose which inactivates rapidly and totally active rat P05121 ( 300 microg/kg/min ) was selected to be tested on the thrombosis model . Infusion of 33H I P08709 beginning 10 min before the ADP application significantly reduced the lysis time in comparison to the control ( 123 +/- 30 s versus 169 +/- 33 s , P < 0.05 , paired Student 's t-test ) and the cumulative thrombus area during the lysis period was decreased by 56 +/- 7 % . These results demonstrate that inactivation of P05121 is able to accelerate lysis of a platelet-rich clot in a mesenteric arteriole of the rat . Thus active P05121 most likely participates to the resistance to thrombolysis in the arteriolar microcirculation and its inactivation may shorten ischemic periods after microvascular obstruction such as e.g. during cerebral stroke . [ Recent advances on the studies of the platelet 's inhibition and aggregation. State of the art of new Q9H244 antagonists ] . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : i ) it is a pro-drug that needs to be metabolized to its active metabolite ; ii ) it has a delayed onset and offset of action ; iii ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG ( Coronary Artery Bypass Grafting ) related bleeding complications . Two direct and reversible Q9H244 antagonists , cangrelor and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 did not prove superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 proved to be superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but was , like prasugrel , was associated with higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery was necessary in ticagrelor-treated patients compared to clopidogrel-treated patients to limit the severity of post-surgical bleeding . DB06441 for patients undergoing percutaneous coronary intervention : evidence from a meta-analysis of randomized trials . DB06441 is a new parenteral adenosine diphosphate Q9H244 receptor inhibitor with rapid , profound and reversible inhibition of platelet activity . The aim of this meta-analysis was to evaluate efficacy and safety of this new agent in patients undergoing percutaneous coronary intervention ( P05154 ) . We searched PubMed , Cochrane Library , EMBASE , Web of Science and CINAHL databases from the inception through April 2013 . Randomized controlled trials ( RCTs ) comparing cangrelor with control ( clopidogrel/placebo ) were selected . We used the random-effects models to calculate the risk ratio . The primary efficacy outcome was risk of myocardial infarction , and the primary safety outcome was TIMI major bleeding at 48 h . Three RCTs included a total of 25,107 participants . Effects of DB06441 were not different against comparators for myocardial infarction ( MI ) ( Risk ratio [ RR ] 0.94 , 95 % confidence interval [ CI ] 0.78-1.13 ) and all-cause mortality ( RR 0.72 , 95 % CI 0.36-1.43 ) . However , cangrelor significantly reduced the risk of ischemia-driven revascularization ( RR 0.72 , 95 % CI 0.52-0.98 ) , stent thrombosis ( RR 0.60 , 95 % CI 0.44-0.82 ) and Q wave MI ( RR 0.53 , 95 % CI 0.30-0.92 ) without causing extra major bleeding ( Thrombolysis in Myocardial infarction criteria ) and severe or life-threatening bleeding ( Global utilization of streptokinase and tissue plasminogen activator for occluded coronary arteries criteria ) . Separate analysis against only clopidogrel also showed similar findings except Q wave MI outcome . Use of cangrelor during P05154 might reduce the risk of ischemia-driven revascularization and stent thrombosis , without causing extra major bleeding . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . Echicetin coated polystyrene beads : a novel tool to investigate GPIb-specific platelet activation and aggregation. P04275 /ristocetin ( P04275 /R ) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin , which also binds P04275 . These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets . Here , we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads , which specifically activate GPIb . We compared platelet activation induced by echicetin beads to P04275 /R . Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling . Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets , while under the same conditions P04275 /R treatment led only to αIIbβ3-independent platelet agglutination . The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation , while the total amount of echicetin used for activation is not critical . Echicetin beads induced strong phosphorylation of several proteins including p38 , P29323 and P31749 . Synergistic signaling via Q9H244 and thromboxane receptor through secreted ADP and TxA2 , respectively , were important for echicetin bead triggered platelet activation . Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation . Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways . Dysfunction of the P19957 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia . Patients with myeloproliferative disorders ( MPDs ) , such as essential thrombocythemia ( ET ) have increased risk of thrombosis and bleeding , which are major sources of morbidity and mortality . Most P53602 patients have a gain of function mutation in O60674 ( JAK2V617F ) , but little is known how JAK2V617F affects platelet function . Here , we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin ( which couples to O60674 ) on this pathway . In contrast , SFLLRN-mediated P16109 expression , DB00171 secretion , phosphorylation of the PKC substrate pleckstrin , and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets . In addition , thrombopoietin-mediated O60674 phosphorylation was unchanged , suggesting that signaling pathways activated downstream of O60674 are impaired . Indeed , we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the P19957 kinase substrate Akt , and have reduced activation of Rap1 in response to thrombopoietin , DB01277 ,ADP , SFLLRN , and thrombin . This effect was independent of Giα Q9H244 purinergic receptor function as ADP-mediated inhibition of P50552 phosphorylation was unchanged . These results demonstrate that the P19957 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients , resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation . Electrocardiographic safety of cangrelor , a new intravenous antiplatelet agent : a randomized , double-blind , placebo- and moxifloxacin-controlled thorough QT study . DB06441 is an intravenous Q9H244 inhibitor under investigation as an antiplatelet drug in the setting of acute coronary syndromes . To determine the electrophysiologic safety of parenteral cangrelor , cardiac repolarization effects were measured in 67 healthy volunteers ( aged 18-45 years ) in a randomized crossover design , including 4 treatment sequences of therapeutic cangrelor , supratherapeutic cangrelor , placebo , and moxifloxacin ( positive control ) . Triplicate electrocardiogram measurements and pharmacokinetic samples were collected at baseline and 9 time points postdose on day 1 . For both cangrelor and moxifloxacin , time-matched , placebo-adjusted change in QT from baseline was evaluated using an individual ( QTcI ) heart rate correction . After cangrelor dosing , change in QTcI was < 5 ms at all times points and all corresponding upper 2-sided 90 % confidence intervals ( CIs ) were < 10 ms . Although moxifloxacin failed to show a lower CI > 5 ms , expected time trends and lower CI > 4.0 ms demonstrate assay sensitivity . QTcI was not affected by plasma concentrations of cangrelor metabolites , and cangrelor had no other adverse effects on electrocardiographic parameters . Clinically , cangrelor exposure was well tolerated . Thus , this thorough QT study demonstrated that therapeutic and supratherapeutic cangrelor doses do not adversely affect cardiac repolarization in normal volunteers ( clinicaltrials.gov ; identifier NCT00699504 ) . N-arachidonoyl-L-serine is neuroprotective after traumatic brain injury by reducing apoptosis . N-arachidonoyl-L-serine ( AraS ) is a brain component structurally related to the endocannabinoid family . We investigated the neuroprotective effects of AraS following closed head injury induced by weight drop onto the exposed fronto-parietal skull and the mechanisms involved . A single injection of AraS following injury led to a significant improvement in functional outcome , and to reduced edema and lesion volume compared with vehicle . Specific antagonists to CB2 receptors , transient receptor potential vanilloid 1 ( Q8NER1 ) or large conductance calcium-activated potassium ( BK ) channels reversed these effects . Specific binding assays did not indicate binding of AraS to the Q9Y2T6 cannabinoid receptor . N-arachidonoyl-L-serine blocked the attenuation in phosphorylated extracellular-signal-regulated kinase 1/2 ( P29323 ) levels and led to an increase in pAkt in both the ipsilateral and contralateral cortices . Increased levels of the prosurvival factor Bcl-xL were evident 24 hours after injury in AraS-treated mice , followed by a 30 % reduction in caspase-3 activity , measured 3 days after injury . Treatment with a CB2 antagonist , but not with a P21554 antagonist , reversed this effect . Our results suggest that administration of AraS leads to neuroprotection via P29323 and Akt phosphorylation and induction of their downstream antiapoptotic pathways . These protective effects are related mostly to indirect signaling via the CB2R and Q8NER1 channels but not through P21554 or Q9Y2T6 receptors . State of the art of new Q9H244 antagonists . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : ( a ) it is a pro-drug that needs to be metabolized to its active metabolite ; ( b ) it has a delayed onset and offset of action and ( c ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and a more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG-related bleeding complications . Two direct and reversible Q9H244 antagonists , DB06441 and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 is not superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 is superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but , like prasugrel , is associated with a higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery is necessary in ticagrelor-treated patients compared to clopiodgrel-treated patients to limit the severity of post-surgical bleeding . Pharmacodynamic effects during the transition between cangrelor and ticagrelor . OBJECTIVES : This study sought to determine pharmacodynamic effects during transition from intravenous cangrelor to oral ticagrelor and from oral ticagrelor to intravenous cangrelor . BACKGROUND : DB06441 is an intravenous antagonist of Q9H244 and its use will require transition to and from oral agents . METHODS : Patients ( n = 12 ) with stable coronary artery disease who were taking aspirin 81 mg daily were recruited . On study day 1 , they received a bolus plus 2-h infusion of cangrelor plus a 180-mg dose of ticagrelor at either 0.5 h ( n = 6 ) or 1.25 h ( n = 6 ) . Pharmacodynamic effects ( light transmission platelet aggregation in response to 20 and 5 μmol/l adenosine diphosphate , VerifyNow , Q9H244 assay ( Accumetrics , San Diego , California ) , vasodilator-stimulated phosphoprotein index , and flow cytometry ) were assessed during and after the cangrelor infusion . Patients took 90 mg of ticagrelor twice daily for either 6 ( n = 6 ) or 7 ( n = 6 ) doses . On study day 5 , pharmacodynamic effects were assessed before and during a bolus plus 2-h infusion of cangrelor . RESULTS : During cangrelor infusion , extensive inhibition of platelet function reflected by limited residual platelet reactivity was apparent . After cangrelor was stopped , the antiplatelet effects of ticagrelor were preserved despite a modest increase in platelet reactivity . CONCLUSIONS : DB08816 given before or during infusion of cangrelor did not attenuate the pharmacodynamic effects of cangrelor . The pharmacodynamic effects of ticagrelor were preserved when ticagrelor was given during infusion of cangrelor . Consistent with the reversible binding of ticagrelor , this oral Q9H244 antagonist can be administered before , during , or after treatment with cangrelor . DB06441 AstraZeneca . AstraZeneca is developing the P2T ( P2YADP ) purinoceptor antagonist and platelet aggregation inhibitor , cangrelor , for the potential treatment of unstable angina and as an ultrafast-acting intravenous antithrombotic agent . It is in phase IIb clinical trials [ 315723 ] . NDA and MAA applications are planned for after 2003 [ 275466 ] , [ 314472 ] . It superseded the earlier compound , ARL-67085 , which also reached phase II trials [ 328760 ] . In ex vivo samples of angina patients ' blood , cangrelor inhibits platelet/monocyte conjugate development , which indicate the drug has some degree of disease-modifying activity [ 377418 ] . AstraZeneca is also developing derivatives of cangrelor . Removal of the triphosphate side chain , modification of the ribose to a carbocycle and the purine to a triazolopyridine resulted in a potent ( IC50 = 4 nM ) orally-active P2T/ Q9H244 receptor antagonist . A lead compound was scheduled to enter trials as an antithrombotic agent in July 2000 [ 377666 ] . In March 1999 , Lehman Brothers predicted a 30 % probability that the drug would reach world markets and would be launched in 2002 [ 336599 ] . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Erosive arthritis and hepatic granuloma formation induced by peptidoglycan polysaccharide in rats is aggravated by prasugrel treatment . Administration of the thienopyridine Q9H244 receptor antagonist , clopidogrel , increased the erosive arthritis induced by peptidoglycan polysaccharide ( PG-PS ) in rats or by injection of the arthritogenic K/BxN serum in mice . To determine if the detrimental effects are caused exclusively by clopidogrel , we evaluated prasugrel , a third-generation thienopyridine pro-drug , that contrary to clopidogrel is mostly metabolized into its active metabolite in the intestine . Prasugrel effects were examined on the PG-PS-induced arthritis rat model . Erosive arthritis was induced in Lewis rats followed by treatment with prasugrel for 21 days . Prasugrel treated arthritic animals showed a significant increase in the inflammatory response , compared with untreated arthritic rats , in terms of augmented macroscopic joint diameter associated with significant signs of inflammation , histomorphometric measurements of the hind joints and elevated platelet number . Moreover , fibrosis at the pannus , assessed by immunofluorescence of connective tissue growth factor , was increased in arthritic rats treated with prasugrel . In addition to the arthritic manifestations , hepatomegaly , liver granulomas and giant cell formation were observed after PG-PS induction and even more after prasugrel exposure . Cytokine plasma levels of P01584 , P05231 , MIP1 alpha , MCP1 , Q16552 and RANTES were increased in arthritis-induced animals . P22301 plasma levels were significantly decreased in animals treated with prasugrel . Overall , prasugrel enhances inflammation in joints and liver of this animal model . Since prasugrel metabolites inhibit neutrophil function ex-vivo and the effects of both clopidogrel and prasugrel metabolites on platelets are identical , we conclude that the thienopyridines metabolites might exert non-platelet effects on other immune cells to aggravate inflammation . P62158 interacts with the platelet ADP receptor P47900 . P47900 [ P2 ( purinergic type-2 ) -receptor 1 ] is a G-protein-coupled ADP receptor that regulates platelet activation and ADP-induced Ca2+ signalling . Studies using P47900 -knockout mice , G(q)-deficient mice or P47900 -selective inhibitors have previously identified a key role for P47900 in pathophysiological thrombus formation at high shear stress . We provide evidence that a positively charged juxtamembrane sequence within the cytoplasmic C-terminal tail of P47900 can bind directly to the cytosolic regulatory protein calmodulin . Deletion by mutagenesis of the calmodulin-binding domain of P47900 inhibits intracellular Ca2+ flux in transfected cells . These results suggest that the interaction of calmodulin with the P47900 C-terminal tail may regulate P47900 -dependent platelet aggregation . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Genetic aspects of ischemic stroke : coagulation , homocysteine , and lipoprotein metabolism as potential risk factors . Stroke is one of the most common causes of death and long term disability throughout the world . It may be the outcome of a number of monogenic disorders or , more commonly , a polygenic multifactorial disease . Numerous studies have investigated the role of genetics in the pathogenesis of ischemic stroke , with varied and often contradictory results . The candidate ' stroke risk ' genes affecting haemostasis ( P12259 , F2 , P02671 / P02675 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / P08514 , P17301 , P07359 , TPA , Q96IY4 , P07204 , PZ , P08758 ) , homocysteine metabolism ( P42898 , P35520 , Q99707 ) , and lipid metabolism ( apo E , P06858 , P11597 , O95477 , apo AI , apo CIII , apo AIV , apo AV , apo B , apo H , apo(a) , P27169 /2/3 , P01130 / P78380 ) are evaluated in this review . By examining meta-analyses and case-control studies , we made a classification of gene/gene polymorphisms according to the degree of association with ischemic stroke risk . The data assembled could be very useful for further meta-analysis and for future clinical applications . Clinical overview of promising nonthienopyridine antiplatelet agents . Three novel nonthienopyridine antiplatelet agents -- cangrelor , ticagrelor ( AZD6140 ) , and P35240 530348 -- are in advanced clinical testing in patients with coronary artery disease . DB06441 and ticagrelor are direct and reversible inhibitors of the platelet adenosine 5'-diphosphate Q9H244 receptor , whereas P35240 530348 is a thrombin receptor antagonist . Clinical data available to date for each of these compounds suggest that they have safety and efficacy profiles that will be advantageous to patients with acute coronary syndromes or undergoing percutaneous intervention . We review the clinical features of these new platelet inhibition therapies . Differential requirements for Th1 and Th17 responses to a systemic self-antigen . T cell- P25054 interactions are essential for the initiation of effector responses against foreign and self-antigens , but the role of these interactions in generating different populations of effector T cells in vivo remains unclear . Using a model of P01730 (+) T cell responses to a systemic self-antigen without adjuvants or infection , we demonstrate that activation of APCs augments Th17 responses much more than Th1 responses . Recognition of systemic Ag induces tolerance in self-reactive P01730 (+) T cells , but induction of P25942 signaling , even under tolerogenic conditions , results in a strong , Ag-specific Q16552 response without large numbers of IFN-γ-producing cells . Transfer of the same P01730 (+) T cells into lymphopenic recipients expressing the self-antigen results in uncontrolled production of Q16552 , IFN-γ , and systemic inflammation . If the Ag-specific T cells lack P29965 , production of Q16552 but not IFN-γ is decreased , and the survival time of recipient mice is significantly increased . In addition , transient blockade of the initial MHC class II-dependent T cell- P25054 interaction results in a greater reduction of Q16552 than of IFN-γ production . These data suggest that Th17 differentiation is more sensitive to T cell interactions with APCs than is the Th1 response , and interrupting this interaction , specifically the P25942 pathway , may be key to controlling Th17-mediated autoimmunity . DB06441 in percutaneous coronary intervention . DB06441 is a novel , intravenous Q9H244 receptor antagonist in development for use in percutaneous coronary intervention . Currently in Phase III testing , the reversible platelet inhibitor provides several inherent advantages over other Q9H244 receptor antagonists in this setting for the prevention of adverse cardiac events . Unlike the class of thienopyridines ( ticlopidine , clopidogrel and potentially soon to be available , prasugrel ) , cangrelor has nearly immediate onset after a bolus dose and a short half-life , and achieves maximal inhibition of ADP-mediated platelet function . DB06441 's distinct mechanism of action allows for intravenous administration and avoids both hepatic and renal metabolism . These unique characteristics make cangrelor a promising agent for use in cardiovascular patients undergoing percutaneous coronary intervention . Potentiation of platelet aggregation by heparin in human whole blood is attenuated by Q9H244 and P47900 antagonists but not aspirin . INTRODUCTION : DB01109 ( UFH ) potentiates platelet aggregation induced by some agonists . Q9H244 and P47900 receptors play a major role in amplifying platelet aggregation . We assessed the ability of cangrelor , a selective Q9H244 antagonist , A2P5P , a selective P47900 antagonist , and aspirin to block the potentiating effects of heparin . MATERIALS AND METHODS : Whole blood from healthy human volunteers was anticoagulated with either hirudin or UFH 10 IU/ml . Some tubes anticoagulated with hirudin also contained UFH 1 or 10 IU/ml . The low-molecular-weight heparin dalteparin was also assessed . Platelet aggregation was performed using whole blood single-platelet counting . Dense granule release was assessed using 14C-5HT-labelled platelets . RESULTS : UFH and , to a lesser extent , dalteparin potentiated platelet aggregation induced by ADP , Q15004 , 5HT , U46619 , epinephrine and TRAP in a concentration-dependent manner but inhibited aggregation induced by collagen . DB06441 effectively opposed the potentiating effects of heparins on sustained aggregation induced by ADP , Q15004 , 5HT , U46619 and TRAP but had less effect on epinephrine-induced aggregation , whereas A2P5P was more effective at blocking both the initial phase of ADP-induced aggregation and the aggregation response to epinephrine , reflecting the differences in G protein coupling between the agonist receptors . DB00945 had no effect on potentiation by heparin . Heparins did not increase ADP- or TRAP-induced 14C-5HT release . CONCLUSIONS : Heparins potentiate platelet responses to ADP and numerous other agonists . This potentiation is attenuated by cangrelor and A2P5P , and is not mediated by increased dense granule release . ADP receptor antagonists but not aspirin may have potential therapeutic benefits in counteracting the pro-thrombotic effects of heparins . Emerging antithrombotic drugs for acute coronary syndrome . INTRODUCTION : Acute coronary syndrome ( ACS ) encompasses acute myocardial infarction ( MI ) and unstable angina . Activation of platelets and coagulation cascade plays a central role in the development of ACS . Over the past decade , there have been substantial improvements in the strategies for secondary prevention of ACS , including the development of more potent oral antiplatelet agents such as prasugrel and ticagrelor . However , therapies with even better efficacy and safety profiles and more rapid onset and offset of action would be desirable . AREAS COVERED : This review discusses the advantages and disadvantages of the currently available antithrombotic agents and describes the findings from recent clinical trials of three novel agents ; cangrelor ( an intravenous Q9H244 receptor antagonist ) , vorapaxar ( protease-activated receptor-1 inhibitor ) and rivaroxaban ( an oral factor Xa inhibitor ) . EXPERT OPINION : DB06441 appears more promising than clopidogrel when a very rapid onset and reversal of antiplatelet effect is needed . DB09030 in addition to standard oral antiplatelet therapy was effective in patients with prior MI , but was not safe in patients with a prior stroke . Low dose rivaroxaban decreased cardiovascular events and mortality in patients post-ACS compared to placebo , although bleeding was increased . DB08816 increases adenosine plasma concentration in patients with an acute coronary syndrome . OBJECTIVES : This study aimed to investigate the impact of ticagrelor on adenosine plasma concentration ( P25054 ) in acute coronary syndrome ( ACS ) patients . BACKGROUND : DB08816 is a direct-acting Q9H244 -adenosine diphosphate receptor blocker . The clinical benefit of ticagrelor compared with clopidogrel in ACS patients suggests that the drug has non-platelet-directed properties . Animal and in vitro models suggested that the " pleiotropic " properties of ticagrelor may be related to an interaction with adenosine metabolism . METHODS : We prospectively randomized 60 ACS patients to receive ticagrelor or clopidogrel . The P25054 was measured by liquid chromatography . To assess the mechanism of P25054 variation , we measured adenosine deaminase concentration , adenosine uptake by red blood cells , and cyclic adenosine monophosphate production by cells overexpressing adenosine receptors . The Q9H244 -adenosine diphosphate receptor blockade was assessed by the vasodilator-stimulated phosphoprotein index . RESULTS : Patients receiving ticagrelor had significantly higher P25054 than patients receiving clopidogrel ( 1.5 μM [ interquartile range : 0.98 to 1.7 μM ] vs. 0.68 μM [ interquartile range : 0.49 to 0.78 μM ] ; p < 0.01 ) . The P25054 was not correlated with vasodilator-stimulated phosphoprotein ( p = 0.16 ) . Serum-containing ticagrelor inhibited adenosine uptake by red blood cells compared with clopidogrel or controls ( p < 0.01 for both comparisons ) . DB00640 deaminase activity was similar in serum of patients receiving clopidogrel or ticagrelor ( p = 0.1 ) . DB08816 and clopidogrel had no direct impact on adenosine receptors ( p = not significant ) . CONCLUSIONS : DB08816 increases P25054 in ACS patients compared with clopidogrel by inhibiting adenosine uptake by red blood cells . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . First report of warfarin dose requirements in patients possessing the P11712 *12 allele . BACKGROUND : DB00682 is the most frequently prescribed anticoagulant in North America and Europe . It is administered as a racemate , but S-warfarin is principally responsible for its anticoagulant activity . Cytochrome P450 ( CYP ) 2C9 is the enzyme primarily responsible for the metabolism of S-warfarin . Numerous variant alleles of P11712 have been identified . The P11712 *12 ( rs9332239 ) allele harbors a P489S substitution in P11712 which has been shown to result in a 40 % decline in catalytic activity in vitro . CASES : Four Caucasian patients with a low mean weekly warfarin dose ( MWWD ) were genotyped for P11712 , Q9BQB6 and P02649 variant alleles . None of the four patients carried the common P11712 variant alleles ( *2 , *3 , *5 , *6 , *7 , *8 , *9 , *11 , *13 ) despite a relatively low MWWD ( 23.4±7.94 mg ) compared to 208 patients carrying the CYP29C9*1 genotype ( 32.2±12.65 mg ) . Given that P11712 *12 confers decreased in vitro activity to the enzyme , we investigated whether these patients carried this allele . All four patients were P11712 *12 CT heterozygotes . Individual comparisons with patients possessing the same Q9BQB6 and P02649 genotypes also demonstrated lower dose requirements in the patients that possessed P11712 *12 allele . CONCLUSIONS : There are no reports of the clinical impact of rs9332239 on P11712 substrates . This is the first report of patients with the rare P11712 *12 genotype and lower warfarin dose requirements . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . The metabolic syndrome - an ongoing story . The metabolic syndrome refers to the clustering of cardiovascular risk factors that include diabetes , obesity , dyslipidaemia and hypertension . Due to various definitions and unexplained pathophysiology it is still a source of medical controversy . P01308 resistance and visceral obesity have been recognized as the most important pathogenic factors . P01308 resistance could be defined as the inability of insulin to produce its numerous actions , in spite of the unimpaired secretion from the beta cells . Metabolic abnormalities result from the interaction between the effects of insulin resistance located primarily in the muscle and adipose tissue and the adverse impact of the compensatory hyperinsulinaemia on tissues that remain normally insulin-sensitive . The clinical heterogeneity of the syndrome can be explained by its significant impact on glucose , fat and protein metabolism , cellular growth and differentiation , and endothelial function . Visceral fat represents a metabolically active organ , strongly related to insulin sensitivity . Moderating the secretion of adipocytokines like leptin , adiponectin , plasminogen activator inhibitor 1 ( P05121 ) , tumor necrosis factor alfa ( P01375 -alfa ) , interleukin-6 ( P05231 ) and resistin , it is associated with the processes of inflammation , endothelial dysfunction , hypertension and atherogenesis . In 2005 , the International Diabetes Federation ( IDF ) has proposed a new definition , based on clinical criteria and designed for global application in clinical practice . Visceral obesity measured by waist circumference is an essential requirement for diagnosis ; other variables include increased triglyceride and decreased HDL levels , hypertension and glucose impairment . Whatever the uncertainties of definition and etiology , metabolic syndrome represents a useful and simple clinical concept which allows earlier detection of type 2 diabetes and cardiovascular disease . DB06441 : review of the drug and the CHAMPION programme ( including PHOENIX ) . Platelet inhibition is the main goal of ancillary pharmacologic therapy during percutaneous coronary interventions ( P05154 ) . Thienopyridines and ticagrelor are oral drugs developed for this purpose . DB06441 is an intravenous , non-thienopyridine antagonist of the Q9H244 receptor with a rapid , potent , predictable , and quickly reversible effect . DB06441 has been studied in a broad population intended to receive P05154 in the CHAMPION program , where it was compared with different clopidogrel regimens . The first two trials , CHAMPION P05154 and PLATFORM , failed their primary objective , likely for challenges in the adjudication of P05154 -related myocardial infarction . In a third trial that implemented the universal definition of MI , CHAMPION PHOENIX , a reduction of thrombotic events , including stent thrombosis , was observed . In the BRIDGE trial cangrelor has been studied in patients who had to prematurely interrupt antiplatelet therapy for surgery . DB06441 appears a promising agent in patients who require P05154 or when a rapid reversal is needed . Pharmacokinetics and pharmacodynamics of a bolus and infusion of cangrelor : a direct , parenteral Q9H244 receptor antagonist . The purpose of this study is to evaluate the safety , tolerability , pharmacokinetics , and pharmacodynamics of cangrelor administered as an intravenous bolus plus a continuous infusion in healthy volunteers . Twenty-two healthy volunteers are randomized to receive 1 of 2 intravenous cangrelor dosing regimens : a 15-microg/kg bolus followed by a 2-microg/kg/min infusion or a 30-microg/kg bolus followed by a 4-microg/kg/min infusion . The infusion is continued for 60 minutes , and serial blood samples are obtained for evaluation of pharmacokinetic and pharmacodynamic parameters . Administration of an intravenous bolus followed by a continuous infusion rapidly achieves maximum concentrations of cangrelor that are associated with extensive platelet inhibition within 2 minutes . Moreover , extensive platelet inhibition is maintained throughout the infusion period with near-full recovery of platelet function within 60 to 90 minutes of terminating the infusion . The effect of high-dose cangrelor is more consistent and demonstrates a greater level of inhibition on adenosine diphosphate-induced P16109 expression ; how ever , no significant differences are observed between the 2 dosing regimens with regard to platelet aggregation or time to recovery of platelet function . DB06441 administered as an intravenous bolus followed by a continuous infusion in healthy volunteers offers rapid and reversible inhibition of platelet function . Contribution of the Q9H244 receptor-mediated pathway to platelet hyperreactivity in hypercholesterolemia . BACKGROUND : In hypercholesterolemia , platelets demonstrate increased reactivity and promote the development of cardiovascular disease . OBJECTIVE : This study was carried out to investigate the contribution of the ADP receptor Q9H244 -mediated pathway to platelet hyperreactivity due to hypercholesterolemia . METHODS : P01130 -deficient mice and C57Bl/6 wild-type mice were fed on normal chow and high-fat ( Western or Paigen ) diets for 8 weeks to generate differently elevated cholesterol levels . Q9H244 receptor-induced functional responses via G(i) signaling were studied ex vivo when washed murine platelets were activated by 2MeSADP and PAR4 agonist AYPGKF in the presence and absence of indomethacin . Platelet aggregation and secretion , α(IIb)β(3) receptor activation and the phosphorylation of extracellular signal-regulated protein kinase ( P29323 ) and Akt were analyzed . RESULTS : Plasma cholesterol levels ranged from 69 ± 10 to 1011 ± 185 mg dL(-1) depending on diet in mice with different genotypes . Agonist-dependent aggregation , dense and α-granule secretion and JON/A binding were gradually and significantly ( P < 0.05 ) augmented at low agonist concentration in correlation with the increasing plasma cholesterol levels , even if elevated thromboxane generation was blocked . These functional responses were induced via increased levels of G(i) -mediated P29323 and Akt phosphorylation in hypercholesterolemic mice vs. normocholesterolemic animals . In addition , blocking of the Q9H244 receptor by AR-C69931MX ( DB06441 ) resulted in strongly reduced platelet aggregation in mice with elevated cholesterol levels compared with normocholesterolemic controls . CONCLUSIONS : These data revealed that the Q9H244 receptor pathway was substantially involved in platelet hyperreactivity associated with mild and severe hypercholesterolemia . DB06441 for treatment during percutaneous coronary intervention . Dual antiplatelet therapy consisting of aspirin and a Q9H244 -receptor antagonist is important for preventing major adverse cardiovascular events in patients managed with percutaneous coronary intervention ( P05154 ) . The current Q9H244 -receptor antagonists are only available for oral administration and exhibit a delayed onset of action . Furthermore , several days are required for platelet function to return to normal following cessation of therapy . DB06441 is an intravenous DB00171 analog that directly , selectively and reversibly inhibits Q9H244 receptors on platelets . A 30-μg/kg bolus dose followed by a 4-μg/kg per minute continuous infusion of cangrelor achieves peak concentration and maximal platelet inhibition within minutes of administration . DB06441 also demonstrates a fast offset as normal platelet function is restored 1-2 h after cessation of the infusion . Three large , double-blind , randomized trials - CHAMPION PLATFORM , CHAMPION P05154 and CHAMPION PHOENIX - assessed the efficacy and safety of cangrelor compared with clopidogrel ( during or immediately after P05154 ) or placebo in the setting of P05154 . In the most recent CHAMPION PHOENIX trial , cangrelor was superior to clopidogrel for preventing adverse cardiovascular events with no significant increase in major bleeding . Based on the clinical trial results combined with unique properties such as intravenous administration and fast onset and offset , cangrelor may provide benefit in certain patients undergoing P05154 . Consequences of the Y139F Vkorc1 mutation on resistance to AVKs : in-vivo investigation in a 7th generation of congenic Y139F strain of rats . OBJECTIVES : In humans , warfarin is used as an anticoagulant to reduce the risk of thromboembolic clinical events . DB00682 derivatives are also used as rodenticides in pest control . The gene encoding the protein targeted by anticoagulants is the Vitamin K-2,3-epoxide reductase subunit 1 ( Q9BQB6 ) . Since its discovery in 2004 , various amino acid and transcription-regulatory altering Q9BQB6 mutations have been identified in patients who required extreme antivitamin K dosages , or wild populations of rodents that were difficult to control with anticoagulant rodenticides . One unresolved question concerns the dependency of the Q9BQB6 on the genetic background in humans and rodents that respond weakly or not at all to anticoagulants . Moreover , an important question requiring further analyses concerns the role of the Vkorc1 gene in mediating resistance to more recently developed warfarin derivatives ( superwarfarins ) . METHODS : In this study , we bred a quasicongenic rat strain by using a wild-caught anticoagulant resistant rat as a donor to introduce the Y > F amino acid change at position 139 in the Vkorc1 into the genetic background of an anticoagulant susceptible Spraque-Dawley recipient strain . RESULTS AND CONCLUSION : In this manuscript we report the prothrombin times measured in the P08709 generation after exposure to chlorophacinone , bromadiolone , difenacoum and difethialone . We observed that the mutation Y139F mediates resistance in an otherwise susceptible genetic background when exposed to chlorophacinone and bromadiolone . However , the physiological response to the super-warfarins , difenacoum and difethialone , may be strongly dependent on other genes located outside the congenic interval ( 28.3 cM ) bracketing the Vkorc1 in our P08709 generation congenic strain . Emerging Q9H244 receptor antagonists : role in coronary artery disease . The use of oral antiplatelet therapy in reducing vascular events has been extensively studied . Currently available oral antiplatelet agents include aspirin and the thienopyridine Q9H244 receptor antagonists . These classes are combined frequently in the setting of acute coronary syndrome and percutaneous coronary intervention ( P05154 ) . Resistance to either or both of these agents is a major concern , as antiplatelet resistance has been linked to an increase in thrombotic events and worse clinical outcomes . As a result , there is a need for newer , more effective antiplatelet agents to address the limitations of currently available therapy . Prasugrel , a third generation thienopyridine , has been approved by both the FDA and European Commission . Two additional Q9H244 agents , ticagrelor and cangrelor are in advanced stages of development . The possible advantages of prasugrel over clopidogrel include a faster onset of action , reduced inter-patient variability and more potent platelet inhibition . DB08816 is an oral reversible Q9H244 antagonist with greater platelet inhibition compared with clopidogrel . DB06441 is being developed as an intravenous Q9H244 antagonist with a very fast onset and offset , which may offer advantages particularly in the setting of coronary intervention . These emerging antiplatelet agents may offer advantages such as faster onset of action , greater potency and reversibility of platelet inhibition . This article summarizes the available clinical data on the upcoming Q9H244 antiplatelet agents in the treatment of coronary artery disease . Cellular mechanisms of the hemostatic effects of desmopressin ( DB00035 ) . The synthetic analog of vasopressin desmopressin ( DB00035 ) is widely used for the treatment of patients with von Willebrand disease ( VWD ) , hemophilia A , several platelet disorders , and uremic bleeding . DB00035 induces an increase in plasma levels of P04275 ( P04275 ) , coagulation factor VIII ( FVIII ) , and tissue plasminogen activator ( t-PA ) . It also has a vasodilatory action . In spite of its extensive clinical use , its cellular mechanism of action remains incompletely understood . Its effect on P04275 and t-PA as well as its vasodilatory effect are likely explained by a direct action on the endothelium , via activation of endothelial vasopressin P30518 receptor and DB02527 -mediated signaling . This leads to exocytosis from Weibel Palade bodies where both P04275 and t-PA are stored , as well as to nitric oxide ( NO ) production via activation of endothelial NO synthase . The mechanism of action of DB00035 on FVIII plasma levels remains to be elucidated . The hemostatic effect of DB00035 likely involves additional cellular effects that remain to be discovered . DB06441 for treatment of coronary thrombosis . OBJECTIVE : To review and assess available literature on the chemistry , pharmacology , pharmacodynamics , pharmacokinetics , clinical studies , adverse events , drug interactions , special populations , and dosing and administration for cangrelor , a product in late stage Phase II clinical trials . DATA SOURCES : A literature search of MEDLINE ( 1966-March 2006 ) , International Pharmaceutical Abstracts ( 1970-February 2006 ) , and Cochrane database ( first quarter 2006 ) was conducted using key terms of cangrelor , AR-C69931MX , and Q9H244 receptor antagonist . Bibliographies of relevant articles were reviewed for additional references . The Medicines Company Web site was reviewed , and a company representative was contacted . STUDY SELECTION AND DATA EXTRACTION : Available English-language literature , including abstracts , preclinical studies , clinical trials , and review articles , was reviewed . DATA SYNTHESIS : DB06441 is a Q9H244 antagonist under development for treatment of acute coronary syndrome . DB06441 has been studied as an intravenous infusion in doses of 2 or 4 microg/kg/min . It inhibits platelet aggregation with rapid onset and offset and does not require metabolism for therapeutic activity . Published Phase II trials have demonstrated safety and inhibition of platelet aggregation . CONCLUSIONS : DB06441 is a promising investigational medication for inhibition of platelet aggregation in acute arterial coronary events . Phase II trials have shown safety and a greater inhibition of platelet aggregation over clopidogrel . Phase III trials will provide more definitive information on clinical efficacy and safety . Until then , the role of cangrelor is uncertain . Efficacy and safety of cangrelor for patients with coronary artery disease : a meta-analysis of four randomized trials . BACKGROUND : The efficacy and safety of new intravenous Q9H244 inhibitor ( cangrelor ) for patients with coronary artery disease ( CAD ) remain unclear . METHODS AND RESULTS : Trials were identified in PubMed , Web of Science , Embase , and Cochrane Database searches . We included four randomized , placebo-controlled reports in the meta-analysis . The database consisted of 36 , 081 patients on cangrelor compared with clopidogrel or placebo . Major adverse cardiac events ( MACE ) were defined as the primary efficacy endpoint and major or severe bleeding at 48 hours was defined as the primary safety endpoint . DB06441 significantly decreased risk of MACE ( OR : 0.87 , P = 0.002 ) and stent thrombosis ( OR : 0.53 , P < 0.001 ) . However , at the same time , an increase in TIMI minor bleeding ( OR : 1.49 , P = 0.04 ) and in GUSTO moderate bleeding ( OR : 1.43 , P = 0.04 ) were observed by cangrelor . CONCLUSIONS : Intravenous administration of cangrelor is benefit to reduce risk of MACE and stent thrombosis in patients with CAD excepting for increased minor bleeding events . DB06441 : a review on pharmacology and clinical trial development . Dual antiplatelet therapy with aspirin and an oral ADP Q9H244 receptor antagonist is the standard-of-care for the prevention of ischemic events in patients with acute coronary syndrome or undergoing percutaneous coronary intervention ( P05154 ) . However , currently available ADP Q9H244 receptor antagonists have several limitations , such as interindividual response variability , drug-drug interactions , slow onset/offset and only oral availability . DB06441 is a reversible , potent , intravenous , competitive inhibitor of the ADP Q9H244 receptor that rapidly achieves near complete and predictable platelet inhibition . Along with reversible binding to the receptor cangrelor also has a very short half-life ( 3-5 min ) , which in turn results in a rapid offset of action . These properties make cangrelor a promising drug for clinical use in patients undergoing P05154 or patients waiting for major surgery but still require antiplatelet protection . This manuscript provides an update of the current status of knowledge on cangrelor , focusing on its pharmacologic properties and clinical trial development , including the BRIDGE and CHAMPION-PHOENIX trials . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . The influence of variation in the Q9H244 receptor gene on in vitro platelet inhibition with the direct Q9H244 antagonist cangrelor . Novel Q9H244 inhibitors are in development to overcome the occurrence of atherothrombotic events associated with poor responsiveness to the widely used Q9H244 inhibitor clopidogrel . DB06441 is an intravenously administered Q9H244 inhibitor that does not need metabolic conversion to an active metabolite for its antiplatelet action , and as a consequence exhibits a more potent and consistent antiplatelet profile as compared to clopidogrel . It was the objective of this study to determine the contribution of variation in the Q9H244 receptor gene to platelet aggregation after in vitro partial Q9H244 receptor blockade with the direct antagonist cangrelor . Optical aggregometry was performed at baseline and after in vitro addition of 0.05 and 0.25 microM cangrelor to the platelet-rich plasma of 254 healthy subjects . Five haplotype-tagging (ht)-SNPs covering the entire Q9H244 receptor gene were genotyped ( rs6798347C > t , rs6787801T > c , rs9859552C > a , rs6801273A > g and rs2046934T > c [ T744C ] ) and haplotypes were inferred . The minor c allele of SNP rs6787801 was associated with a 5 % lower 20 microM ADP-induced peak platelet aggregation ( 0.05 microM cangrelor , p < 0.05 ) . Aa homozygotes for SNP rs9859552 showed 20 % and 17 % less inhibition of platelet aggregation with cangrelor when compared to CC homozygotes ( 0.05 and 0.25 microM cangrelor respectively ; p < 0.05 ) . Results of the haplotype analyses were consistent with those of the single SNPs . Polymorphisms of the Q9H244 receptor gene contribute significantly to the interindividual variability in platelet inhibition after partial in vitro blockade with the Q9H244 antagonist cangrelor . Vascular transcriptional alterations produced by juvenile obesity in Ossabaw swine . We adopted a transcriptome-wide microarray analysis approach to determine the extent to which vascular gene expression is altered as a result of juvenile obesity and identify obesity-responsive mRNAs . We examined transcriptional profiles in the left anterior descending coronary artery ( LAD ) , perivascular fat adjacent to the LAD , and descending thoracic aorta between obese ( n = 5 ) and lean ( n = 6 ) juvenile Ossabaw pigs ( age = 22 wk ) . Obesity was experimentally induced by feeding the animals a high-fat/high-fructose corn syrup/high-cholesterol diet for 16 wk . We found that expression of 189 vascular cell genes in the LAD and expression of 165 genes in the thoracic aorta were altered with juvenile obesity ( false discovery rate ≤ 10 % ) with an overlap of only 28 genes between both arteries . Notably , a number of genes found to be markedly upregulated in the LAD of obese pigs are implicated in atherosclerosis , including P13686 , P61626 , O95715 , P02649 , Q13093 , P17931 , P10451 , P05107 , P04839 , and Q9H244 . Furthermore , pathway analysis revealed the induction of proinflammatory and pro-oxidant pathways with obesity primarily in the LAD . Gene expression in the LAD perivascular fat was minimally altered with juvenile obesity . Together , we provide new evidence that obesity produces artery-specific changes in pretranslational regulation with a clear upregulation of proatherogenic genes in the LAD . Our data may offer potential viable drug targets and mechanistic insights regarding the molecular precursors involved in the origins of overnutrition and obesity-associated vascular disease . In particular , our results suggest that the oxidized LDL/ P78380 /NF-κB signaling axis may be involved in the early initiation of a juvenile obesity-induced proatherogenic coronary artery phenotype . Dysregulation of leukocyte gene expression in women with medication-refractory depression versus healthy non-depressed controls . BACKGROUND : Depressive Disorders ( DD ) are a great financial and social burden . Females display 70 % higher rate of depression than males and more than 30 % of these patients do not respond to conventional medications . Thus medication-refractory female patients are a large , under-served , group where new biological targets for intervention are greatly needed . METHODS : We used real-time quantitative polymerase chain reaction ( qPCR ) to evaluate mRNA gene expression from peripheral blood leukocytes for 27 genes , including immune , Q9Y251 -axis , ion channels , and growth and transcription factors . Our sample included 23 females with medication refractory DD : 13 with major depressive disorder ( MDD ) , 10 with bipolar disorder ( BPD ) . Our comparison group was 19 healthy , non-depressed female controls . We examined differences in mRNA expression in DD vs. controls , in MDD vs. BPD , and in patients with greater vs. lesser depression severity . RESULTS : DD patients showed increased expression for P22301 , P05231 , P30559 , Q99572 , P47900 , and Q8NER1 . BPD patients showed increased P05067 , P16220 , P19838 , P04150 , and P09486 and decreased P01375 expression . Depression severity was related to increased P22301 , P47900 , P51575 , and Q9HBA0 expression . CONCLUSIONS : These results support prior findings of dysregulation in immune genes , and provide preliminary evidence of dysregulation in purinergic and other ion channels in females with medication-refractory depression , and in transcription and growth factors in those with BPD . If replicated in future research examining protein levels as well as mRNA , these pathways could potentially be used to explore biological mechanisms of depression and to develop new drug targets . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . DB06441 : a novel Q9H244 receptor antagonist . Antiplatelet therapy is critical in the prevention of thrombotic complications of acute coronary syndrome and percutaneous coronary interventions . Current antiplatelet agents ( aspirin , clopidogrel and glycoprotein IIb/IIIa antagonists ) have demonstrated the capacity to reduce major adverse cardiac events . However , these agents have limitations that compromise their clinical utility . The platelet Q9H244 receptor plays a central role in platelet function and is a focus in the development of antiplatelet therapies . DB06441 is a potent , competitive inhibitor of the Q9H244 receptor that is administered by intravenous infusion and rapidly achieves near complete inhibition of ADP-induced platelet aggregation . This investigational drug has been studied for use during coronary procedures and the management of patients experiencing acute coronary syndrome and is undergoing evaluation for use in the prevention of perioperative stent thrombosis . The ' allosteric modulator ' P35240 -202676 disrupts G protein-coupled receptor function via sulphydryl-sensitive mechanisms . 1. Previous studies suggest that the thiadiazole compound P35240 -202676 ( N-(2,3-diphenyl-1,2,4-thiadiazol-5-(2H)-ylidene)methanamine ) acts as an allosteric modulator of a variety of structurally distinct G protein-coupled receptors ( GPCRs ) . It was postulated that P35240 -202676 would directly bind a structural motif in the receptor molecule common to divergent members of the GPCR family . The molecular mechanisms of such a promiscuous action , however , remain obscure . 2 . To clarify the mechanism of P35240 -202676 action , we used the functional approach of [35S]GTPgammaS autoradiography with rat brain cryostat sections together with classical membrane [35S]GTPgammaS binding assays to evaluate how the thiadiazole affects G protein activity mediated by various receptors linked to the Gi-family of G proteins . 3 . We found that in the absence of dithiotreitol ( DTT ) , P35240 -202676 ( 10(-7)-10(-5) M ) elicits nonspecific effects in the [35S]GTPgammaS-based G protein activation assays , thereby severely compromising interpretations on the compounds ability to allosterically inhibit receptor-mediated G protein activity . Such a nonspecific behaviour was fully reversed upon addition of DTT ( 1 mM ) , revealing thiol-based mechanism of action . 4 . In routine incubations containing DTT , P35240 -202676 had no effect on receptor-driven G protein activity , as assessed for adenosine A1 , alpha2-adrenergic , cannabinoid P21554 , lysophosphatidic acid Q92633 , muscarinic M2/M4 , purinergic Q9H244 or sphingosine 1-phosphate receptors , suggesting that the thiadiazole does not act as an allosteric modulator of GPCR function . 5 . 1H NMR analysis indicated that P35240 -202676 underwent structural changes after incubation with the reducing agent DTT or with brain tissue . 6 . We conclude that P35240 -202676 modulates GPCRs via thiol modification rather than via true allosteric mechanisms . DB06441 for treatment of arterial thrombosis . INTRODUCTION : Percutaneous coronary intervention ( P05154 ) is a highly effective treatment for obstructive coronary artery disease . Oral platelet Q9H244 receptor antagonists reduce ischemic events in patients treated with P05154 . However , there are several limitations to their use , including variable pharmacodynamics , a slow onset and offset , and in those patients who are pretreated but subsequently require cardiac surgery , increased bleeding . DB06441 is an intravenous agent that provides rapid and intensive inhibition of the Q9H244 receptor that quickly dissipates after discontinuation . A recent , Phase III randomized clinical trial of P05154 patients demonstrated that cangrelor bolus and infusion reduced ischemic events compared with conventional clopidogrel therapy without increasing major bleeding . AREAS COVERED : This review outlines the pharmacodynamics , pharmacokinetics , and the safety and efficacy of cangrelor for the acute treatment of patients undergoing planned P05154 . EXPERT OPINION : DB06441 is an important addition to the current armamentarium of platelet inhibitors as it significantly reduces periprocedural myocardial infarction and stent thrombosis in a broad spectrum of patients , without increasing major bleeding or the need for transfusion . DB06441 will have particular benefit in clopidogrel-naïve patients with high anatomical complexity and/or increased clinical risk ( where the absolute risk for thrombotic and ischemic complications of P05154 is greatest ) . New frontiers in the management of acute coronary syndromes : cangrelor and elinogrel . The activation and aggregation of platelets at sites of vascular injury or near to implanted stent are pivotal in the development of thrombotic events during and after an acute coronary syndrome ( ACS ) or a percutaneous coronary intervention ( P05154 ) . For that reason , an exclusively oral dual antiplatelet treatment regimen with platelet Q9H244 receptor antagonists in addition to the cyclooxygenase inhibitor aspirin has become the cornerstone of treatment in that contest . However , every trial underlines the same problem : if maximizing antiplatelet therapy significantly attenuates ischemic events in patients with coronary artery disease , on the other side it may also increase bleeding phenomena . These limitations have prompted a search for novel antiplatelet agents with a more favorable risk-benefit ratio . Moreover , an early onset of action is desirable during P05154 and an early offset after bleeding events . Two novel antiplatelet agents , DB06441 and Elinogrel , are available in intravenous form ( Elinogrel also in oral form ) and expand this context . Recent trials have tested them against DB00758 regarding efficacy and safety outcomes.This review aimed at providing an overview on intravenous emerging compounds and recent patents in the setting of ACS and P05154 . Catalog of 178 variations in the Japanese population among eight human genes encoding G protein-coupled receptors ( GPCRs ) . We screened DNAs from 48 Japanese individuals for single-nucleotide polymorphisms ( SNPs ) in eight genes encoding G protein-coupled receptors ( GPCRs ) by directly sequencing the entire relevant genomic regions except for repetitive-sequence elements . This approach identified 147 SNPs and 31 insertion/deletion polymorphisms among the eight GPCR genes . On average , we identified one SNP in every 584 nucleotides . Of the 147 SNPs , 69 were identified in P30556 , 12 in P50052 , nine in P35414 , 20 in P37288 , nine in P30518 , 16 in P21728 , six in P08514 , and six in P43119 . Twenty-one SNPs were located in 5' flanking regions , 76 in introns , 32 in exons , and 18 in 3' flanking regions . These variants should contribute to investigations of possible correlations between genotypes and phenotypes as regards susceptibility to disease or responsiveness to drug therapy . Transitioning patients from cangrelor to clopidogrel : pharmacodynamic evidence of a competitive effect . BACKGROUND : DB06441 is a direct , parenteral , and reversible inhibitor of the platelet Q9H244 receptor currently undergoing Phase III testing . As many individuals treated acutely with cangrelor will often be treated long-term with a thienopyridine , it is important to determine the effects of concurrent cangrelor and clopidogrel administration . METHODS AND RESULTS : Ten healthy volunteers received a 600 mg oral loading dose of clopidogrel and then underwent serial platelet function monitoring for 6 h . Two weeks later these same individuals received a 600 mg clopidogrel loading dose simultaneously with a cangrelor IV bolus ( 30 microg/kg ) and a 2-hour infusion ( 4 microg/kg/min ) . A separate group of ten volunteers received a 600 mg clopidogrel loading dose after administration of a cangrelor bolus and a 1-hour infusion . The effects on ADP-induced platelet activation and aggregation were evaluated by flow cytometry , whole-blood electrical impedance , and light-transmittance aggregometry . DB06441 and clopidogrel alone achieved the expected levels of platelet inhibition . However , the sustained platelet inhibition anticipated for clopidogrel treatment did not occur when cangrelor was initiated simultaneously . No such effect was found when clopidogrel was started upon completion of the cangrelor infusion . CONCLUSION : To achieve sustained platelet Q9H244 inhibition in patients treated with cangrelor , clopidogrel administration should be started when the cangrelor infusion is terminated . A critical appraisal of the functional evolution of Q9H244 antagonists as antiplatelet drugs . Q9H244 receptor mediated inhibition of platelet aggregation is one of the most explored and exploited pathways in antiplatelet drug therapy to prevent ischemic events in patients undergoing percutaneous coronary intervention ( P05154 ) for the treatment of the acute coronary syndrome ( ACS ) . DB00208 , DB00758 , Prasugrel , DB08816 , DB06441 and Elinogrel are the Q9H244 inhibitors that act as antiplatelet drugs . In this review , the features of these drugs and the factors reported to be responsible for drug resistance or drug ineffectiveness were described . The features like drug metabolism , reversible or irreversible binding of drugs to their target protein and the mode of administration were observed to evolve along with the antiplatelet drugs . These features also include the drug-drug interactions , the pharmacogenetics and pharmacodynamics of Q9H244 inhibitors . We attempted to critically analyze how the desirable features were met by the Q9H244 inhibitors in the course of time . This review provides an overview of the evolution of Q9H244 inhibitors and may guide the researchers to develop better antiplatelet drugs in the future . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . O60603 stimulation of platelets is mediated by purinergic P51575 -dependent Ca2+ mobilisation , cyclooxygenase and purinergic P47900 and Q9H244 receptor activation . O60603 ( O60603 ) , which recognise and respond to conserved microbial pathogen-associated molecular patterns , is expressed on the platelet surface . Furthermore , it has recently been shown that the O60603 /1 agonist Pam3CSK4 stimulates platelet activation . The aim of the present study was to clarify important signalling events in Pam3CSK4-induced platelet aggregation and secretion . Platelet interaction with Pam3CSK4 and the O60603 /6 agonist MALP-2 was studied by analysing aggregation , DB00171 -secretion , [Ca2+]i mobilisation and thromboxane B2 ( TxB2 ) production . The results show that Pam3CSK4 but not MALP-2 induces [Ca2+]i increase , TxB2 production , dense granule secretion and platelet aggregation . Preincubation of platelets with MALP-2 inhibited the Pam3CSK4-induced responses . The DB00171 -secretion and aggregation in Pam3CSK4-stimulated platelets was impeded by the purinergic P51575 inhibitor P59665 2159 , the purinergic P47900 and Q9H244 antagonists P59665 2179 and cangrelor , the phospholipase C inhibitor U73122 , the calcium chelator BAPT-AM and aspirin . The calcium mobilisation was lowered by P59665 2159 , aspirin and U73122 whereas the TxB2 production was antagonised by P59665 2159 , aspirin and BAPT-AM . When investigating the involvement of the myeloid differentiation factor-88 ( MyD88 ) -dependent pathway , we found that platelets express MyD88 and interleukin 1 receptor-associated kinase ( P51617 ) , which are proteins important in TLR signalling . However , Pam3CSK4 did not stimulate a rapid ( within 10 minutes ) phosphorylation of P51617 in platelets . In conclusion , the results show that Pam3CSK4-induced platelet aggregation and secretion depends on a P51575 -mediated Ca2+ mobilisation , production of TxA2 and ADP receptor activation . The findings in this study further support a role for platelets in sensing bacterial components . Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells . BACKGROUND : Decreased endothelial nitric oxide ( NO ) synthase ( P29474 ) activity and NO production are critical contributors to the endothelial dysfunction and vascular complications observed in many diseases , including diabetes mellitus . Extracellular nucleotides activate P29474 and increase NO generation ; however , the mechanism of this observation is not fully clarified . METHODS AND RESULTS : To elucidate the signaling pathway(s) leading to nucleotide-mediated P29474 phosphorylation at DB00133 -1177 , human umbilical vein endothelial cells were treated with several nucleotides , including DB00171 , UTP , and ADP , in the presence or absence of selective inhibitors . These experiments identified P47900 , P41231 , and possibly P51582 as the purinergic receptors involved in P29474 phosphorylation and demonstrated that this process was adenosine independent . Nucleotide-induced P29474 phosphorylation and activity were inhibited by BAPTA-AM ( an intracellular free calcium chelator ) , rottlerin ( a protein kinase Cdelta inhibitor ) , and protein kinase Cdelta siRNA . In contrast , blockade of AMP-activated protein kinase , calcium/calmodulin-dependent kinase II , calcium/calmodulin-dependent kinase kinase , serine/threonine protein kinase B , protein kinase A , extracellular signal-regulated kinase 1/2 , and p38 mitogen-activated protein kinase did not affect nucleotide-mediated P29474 phosphorylation . CONCLUSIONS : The present study indicates that extracellular nucleotide-mediated P29474 phosphorylation is calcium and protein kinase Cdelta dependent . This newly identified signaling pathway opens new therapeutic avenues for the treatment of endothelial dysfunction . HepG2 human hepatoma cells express multiple cytokine genes . Although cytokines are known to be involved in the regulation of a variety of hepatocellular functions , hepatocytes themselves are generally considered only targets but not producers of these important mediators . In order to investigate whether cells of hepatocellular linages are a potential source of various regulatory cytokines we have estimated the multiple cytokine gene expression in the culture of well differentiated human HepG2 hepatoma cells using RT-PCR . Our findings demonstrate that HepG2 cells express mRNAs for interferon gamma ( P01579 ) , tumour necrosis factor alpha ( P01375 ) , transforming growth factor beta ( TGF-beta ) , macrophage colony-stimulating factor ( P09603 ) , oncostatin-M ( P13725 ) , intercellular adhesion molecule ( P05362 ) , interleukin 4 ( P05112 ) , P05113 , P13232 , P22301 , IL-11 , IL-12 and P05231 receptor ( IL-6R ) . At the same time the expression of IL-1 , P60568 , P08700 , P05231 , P29965 and IL-2R genes was not detected . It was concluded that hepatocytes are potential producers of a variety of cytokines , some of them being able to regulate hepatocellular functions directly , while others are important regulators of leukocyte activity . Thus , on the one hand , hepatocytes may express autoregulatory cytokines and on the other hand , influence the functions of other liver cells like Kupffer , Ito or endothelial cells . Due to their large amount , liver parenchymal cells could be an important source of sytemically acting pro- and anti-inflammatory and other regulatory cytokines . Modulation of the P22301 /IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis . IFN-beta has been shown to be effective in the treatment of multiple sclerosis ( MS ) . However , the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear . Recent studies indicate that under defined conditions , IFN-beta may downregulate DC expression of IL-12 . We and others have shown that IFN-beta may also downregulate P22301 . In light of the recently proposed paradigm that an P22301 /IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease , we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis ( EAE ) , an animal model sharing many features with MS . SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein ( PLP ) , and at onset of EAE were treated every other day with IFN-beta . After eight weeks of treatment , we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of P01579 , P60568 , P05112 , or P05113 in response to the priming determinant used to initiate disease . However , in IFN-beta treated mice , the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of P22301 and a concurrent decreased production of IL-12 . Moreover , the in vivo modulation of the P22301 /IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading . Our results indicate that IFN-beta induces a reciprocal modulation of the P22301 /IL-12 cytokine circuit in vivo . This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading . New Q9H244 blockers . A number of new antiplatelet agents currently in development are anticipated to improve clinical outcomes and safety benefits in patients with acute coronary syndrome ( ACS ) . This article reviews the pharmacology and clinical development of three of these agents : prasugrel , cangrelor , and ticagrelor . Prasugrel , a third-generation , oral thienopyridine , has been shown to be superior to clopidogrel , the current gold standard , in preventing ischemic events in patients with ACS undergoing percutaneous coronary intervention ( P05154 ) , although the bleeding rate was higher . DB06441 , a chemical analog of adenosine triphosphate , is a potent direct platelet Q9H244 antagonist . In development as an intravenous agent , cangrelor is currently being evaluated in two phase III studies in patients requiring P05154 . DB08816 is the first of a new class of orally available antiplatelet agents antagonizing the effects of ADP mediated by Q9H244 ; it is currently being studied in a phase III trial in patients with ACS . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Q9H244 inhibitors : pharmacologic mechanism and clinical relevance . Platelets play a critical role in the pathogenesis of atherothrombotic processes and inhibition of platelet aggregation by antiplatelet therapy is essential and really important in the acute coronary syndromes or in the setting of percutaneous coronary intervention . The first family of adenosine diphosphate Q9H244 receptors inhibiting drug is represented by thienopyridines and among these ticlopidine was the first approved by Food and Drug Administration ; actually its use is discouraged because of its potential side effects ( neutropenia , anemia , gastrointestinal distress and thrombotic thrombocytopenic purpura ) . The second generation of thienopyridines is represented by clopidogrel that has replaced ticlopidine in the clinical practice ; clopidogrel has the largest clinical experience . Prasugrel represents the third generation . It inhibits platelet aggregation by irreversibly blocking the adenosine diphosphate Q9H244 receptor . DB08816 , DB06441 and Enilogrel represent the last generation of thienopyridines . This review is focused on the effects of adenosine diphosphate Q9H244 inhibitors .

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Induction and expression of beta-calcitonin gene-related peptide in rat T lymphocytes and its significance . Our previous data have shown that rat lymphocytes can synthesize calcitonin gene-related peptide ( P80511 ) , a neuropeptide . In this study the type , characteristics , and functional role of lymphocyte-derived P80511 were investigated . The results showed that treatment with Con A ( 4 microg/ml ) and recombinant human P60568 ( rhIL-2 ; 750 U/ml ) for 3-5 days induced P80511 synthesis and secretion by lymphocytes from both thymus and mesenteric lymph nodes in a time-dependent manner . Stimulation of these cells with Con A ( 1-8 microg/ml ) or rhIL-2 ( 94-1500 U/ml ) for 5 days induced a significant increase in P80511 secretion in a concentration-dependent manner . The maximal secretion of P80511 with Con A by thymocytes was elevated from 104+/-11 to 381 +/- 44 pg/10(8) cells , and that by mesenteric lymph node lymphocytes was elevated from 83+/-10 to 349+/-25 pg/10(8) cells , respectively . The maximal P80511 secretion with rhIL-2 by thymocytes was elevated from 116+/-3 to 607+/-23 pg/10(8) , and that by mesenteric lymph node lymphocytes was elevated from 117+/-9 to 704+/- 37 pg/10(8) cells , respectively . The nucleotide sequencing study showed that lymphoid cells expressed beta- P80511 cDNA only . The levels of beta- P80511 mRNA in mitogen-stimulated lymphocytes of both sources were also increased . However , LPS had no such effect on either source of cells . hCGRP(8-37) ( 2.0 microM ) , a P80511 (1) receptor antagonist , enhanced Con A-induced proliferation and P60568 release of thymocytes by 41.3 and 35.8 % over those induced by Con A alone , respectively . The data suggest that T lymphocyte mitogens can induce the production of endogenous beta- P80511 from T lymphocytes , which may partially inhibit the proliferation and P60568 release of rat T lymphocyte under immune challenges . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . [ Changes of protein expression in HepG2 cells with P24941 RNA interference ] . OBJECTIVE : To investigate the effects of stable transfection of P24941 siRNA on biological activities and nuclear proteins of human hepatocellular carcinoma HepG2 cells . METHODS : HepG2 cells were transfected with the eukaryotic expression vector of P(Genesil-1- P24941 ) ; via RNA interference and selected for the ones with stable transfection . We observed the changes in the cell growth curve and cell cycle . The mRNA contents of P24941 and differentially expressed nucleoproteins were detected and analyzed by RT-PCR and two-dimensional ( 2D ) electrophoresis-mass spectrum ( MS ) -database , respectively . Western blotting were used to confirm the differential protein expressions . RESULTS : Compared with P(HK-siRNA);-HepG2 and untransfected groups , the proliferation of HepG2 cells in P( P24941 -siRNA);-HepG2 group was significantly inhibited ( P < 0.01 ) , and the expression of P24941 mRNA significantly decreased in P( P24941 -siRNA);-HepG2 group . Four proteins not expressed in P( P24941 -siRNA);-HepG2 cells were detected by 2D electrophoresis-MS , and they were further confirmed by Western blotting . CONCLUSION : P24941 siRNA significantly suppressed P24941 mRNA expression and the proliferation of HepG2 cells , four proteins not expressed in p( P24941 -siRNA);-HepG2 cells are similar to ribosomal protein P28222 , β-actin , zine finger 276 and chaperonin 10 related protein . Efficacy , safety and tolerability of oral eletriptan in the acute treatment of migraine : results of a phase III , multicentre , placebo-controlled study across three attacks . The efficacy , safety and tolerability of the P28222 /D receptor agonist eletriptan ( 40 mg and 80 mg ) in acute treatment of migraine was evaluated in a multinational , randomized , double-blind , parallel-group , placebo-controlled , three-attack study treating 1153 patients . In the initial attack , significantly more eletriptan patients reported headache relief and complete pain relief at 2 h vs. placebo ( 40 mg 62 % and 32 % , 80 mg 65 % and 34 % , placebo 19 % and 3 % ; P < 0.0001 ) . Headache relief occurred faster after eletriptan , with more patients at both doses reporting relief 30 min ( P < 0.01 ) and 1 h ( P < 0.0001 ) after treatment than after placebo . There was a significantly lower recurrence rate with eletriptan 80 mg compared with placebo ( P < 0.01 ) . Adverse events for all treatments were generally mild or moderate and self-limiting . DB00216 40 mg and eletriptan 80 mg both appear to be effective and well-tolerated acute migraine treatments . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Characterisation of the 5-HT receptor binding profile of eletriptan and kinetics of [3H]eletriptan binding at human P28222 and P28221 receptors . The affinity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] -1H-indole ) for a range of 5-HT receptors was compared to values obtained for other P28222 /1D receptor agonists known to be effective in the treatment of migraine . DB00216 , like sumatriptan , zolmitriptan , naratriptan and rizatriptan had highest affinity for the human P28222 , P28221 and putative 5-ht1f receptor . Kinetic studies comparing the binding of [3H]eletriptan and [3H]sumatriptan to the human recombinant P28222 and P28221 receptors expressed in HeLa cells revealed that both radioligands bound with high specificity ( > 90 % ) and reached equilibrium within 10-15 min . However , [3H]eletriptan had over 6-fold higher affinity than [3H]sumatriptan at the P28221 receptor ( K(D) ) : 0.92 and 6.58 nM , respectively ) and over 3-fold higher affinity than [3H]sumatriptan at the P28222 receptor ( K(D) : 3.14 and 11.07 nM , respectively ) . Association and dissociation rates for both radioligands could only be accurately determined at the P28221 receptor and then only at 4 degrees C . At this temperature , [3H]eletriptan had a significantly ( P < 0.05 ) faster association rate ( K(on) 0.249 min(-1) nM(-1) ) than [3H]sumatriptan ( K(on) 0.024 min(-1) nM(-1) ) and a significantly ( P < 0.05 ) slower off-rate ( K(off) 0.027 min(-1) compared to 0.037 min(-1) for [3H]sumatriptan ) . These data indicate that eletriptan is a potent ligand at the human P28222 , P28221 , and 5-ht1f receptors and are consistent with its potent vasoconstrictor activity and use as a drug for the acute treatment of migraine headache . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . [ The cluster headache : a clinical model of immunologic receptor pathology ? ] . It is well established that cluster headache shows impaired functions at their neuroimmunomodulatory system level . Defect in receptor expression for 5-HT , IL-1 and P60568 have been found in these patients . Sumatriptan , a molecule with agonistic activity for P28221 receptor , truncates cluster headache attacks in 74 % of patients . Flow cytometric analysis of monocytes expressing 5-HT receptor in cluster headache patients showed different trends clearly correlated with the clinical response to sumatriptan . Our findings strongly support the concept that cluster headache patients who are non responders to sumatriptan could present a block in their 5-HT receptor possibly due to specific autoantibodies for this receptor site . Serotonergic regulation of somatosensory cortical development : lessons from genetic mouse models . Monoaminergic neurotransmitter systems appear early during embryogenesis , suggesting that they could play important roles in brain development . Accumulated evidence indicates that serotonin ( 5-hydroxytryptamine , 5-HT ) regulates neural as well as nonneural development , including early aspects of embryonic development , differentiation of neuronal progenitors , and morphogenesis of the craniofacial region , heart and limb . Recent studies using monoamine oxidase-A ( P21397 ) , 5-HT transporter , vesicular monoamine transporter-2 ( Q05940 ) and P28222 receptor single , double and triple knockout mice have provided evidence that the serotonergic system plays important roles in barrel field formation in the developing somatosensory cortex . Here we review evidence from these genetic mouse models and , based on the accumulated evidence , propose a testable model for future studies of mechanisms underlying serotonergic regulation of cortical development . Intrinsic choroidal neurons in the chicken eye : chemical coding and synaptic input . Intrinsic choroidal neurons ( ICNs ) exist in some primates and bird species . They may act on both vascular and non-vascular smooth muscle cells , potentially influencing choroidal blood flow . Here , we report on the chemical coding of ICNs and eye-related cranial ganglia in the chicken , an important model in myopia research , and further to determine synaptic input onto ICN . Chicken choroid , ciliary , superior cervical , pterygopalatine , and trigeminal ganglia were prepared for double or triple immunohistochemistry of calcitonin gene-related peptide ( P80511 ) , choline acetyltransferase ( P28329 ) , dopamine-beta-hydroxylase , galanin ( GAL ) , neuronal nitric oxide synthase ( P29475 ) , somatostatin ( Q8TE85 ) , tyrosine hydroxylase ( TH ) , vasoactive intestinal polypeptide ( P01282 ) , vesicular monoamine-transporter 2 ( Q05940 ) , and alpha-smooth muscle actin . For documentation , light , fluorescence , and confocal laser scanning microscopy were used . Chicken ICNs express P29475 / P01282 /GAL and do not express P28329 and Q8TE85 . ICNs are approached by TH/ Q05940 - , P80511 - , and P28329 -positive nerve fibers . About 50 % of the pterygopalatine ganglion neurons and about 9 % of the superior cervical ganglion neurons share the same chemical code as ICN . Q8TE85 -positive neurons in the ciliary ganglion are GAL/NOS negative . P80511 -positive neurons in the trigeminal ganglion lack GAL/ Q8TE85 . The neurochemical phenotype and synaptic input of ICNs in chicken resemble that of other bird and primate species . Because ICNs lack cholinergic markers , they can not be readily incorporated into current concepts of the autonomic nervous system . The data obtained provide the basis for the interpretation of future functional experiments to clarify the role of these cells in achieving ocular homeostasis . DB00563 induces apoptosis through p53/ P38936 -dependent pathway and increases P12830 expression through downregulation of HDAC/ Q15910 . DB00563 ( MTX ) is a dihydrofolate reductase ( P00374 ) inhibitor widely used as an anticancer drug in different kinds of human cancers . Here we investigated the anti-tumor mechanism of MTX against non-small cell lung cancer ( NSCLC ) A549 cells . MTX not only inhibited in vitro cell growth via induction of apoptosis , but also inhibited tumor formation in animal xenograft model . RNase protection assay ( RPA ) and RT-PCR demonstrated its induction of p53 target genes including DR5 , P38936 , Puma and Noxa . Moreover , MTX promoted p53 phosphorylation at Ser15 and acetylaion at Lys373/382 , which increase its stability and expression . The apoptosis and inhibition of cell viability induced by MTX were dependent on p53 and , partially , on P38936 . In addition , MTX also increased P12830 expression through inhibition of histone deacetylase ( HDAC ) activity and downregulation of polycomb group protein enhancer of zeste homologue 2 ( Q15910 ) . Therefore , the anticancer mechanism of MTX acts through initiation of p53-dependent apoptosis and restoration of P12830 expression by downregulation of HDAC/ Q15910 . Porcine carotid vascular effects of eletriptan ( UK-116,044 ) : a new P28222 /1D receptor agonist with anti-migraine activity . It has been suggested that opening of cephalic arteriovenous anastomoses may be involved in the headache phase of migraine . Indeed , a number of acutely acting anti-migraine drugs , including the ergot alkaloids and sumatriptan , constrict porcine carotid arteriovenous anastomoses . In this study , using pentobarbital anaesthetised pigs , we investigated the effects of eletriptan , a close structural analogue of sumatriptan , on the distribution of common carotid artery blood flow into arteriovenous anastomotic and nutrient ( capillary ) fractions . DB00216 ( 10 , 30 , 100 , 300 and 1000 microg kg(-1) , i.v. ) decreased the total carotid blood flow , exclusively by decreasing cephalic arteriovenous anastomotic blood flow ; nutrient blood flow , particularly to the ear , skin and fat , was significantly increased . The doses of eletriptan needed to reduce arteriovenous anastomotic blood flow and conductance by 50 % ( ED50 ) were , respectively , 117+/-21 microg kg(-1) ( 251+/-45 nmol kg(-1) ) and 184+/-42 microg kg(-1) ( 396+/-91 nmol kg(-1) ) ; the highest dose caused reductions of 84+/-3 % and 77+/-4 % , respectively . The eletriptan-induced changes in carotid haemodynamics were clearly attenuated by pretreating the pigs with the selective P28222 /1D receptor antagonist GR127935 ( 0.5 mg kg(-1) ) . On the basis of these results , we conclude that ( 1 ) the eletriptan-induced constriction of cephalic arteriovenous anastomoses as well as the arteriolar dilatation in head tissues is predominantly mediated by P28222 /1D receptors , and ( 2 ) eletriptan should be effective in aborting migraine headache . Clinical studies have already demonstrated its therapeutic action in migraine patients . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Antibody to ras proteins in patients with colon cancer . The current study examined sera from 160 colon cancer patients and 60 normal individuals to determine whether antibody to mutated P38936 ras protein was present . Studies focused on the aspartic acid substitution at amino acid position 12 ( denoted D12 ) , one of the most common mutations in colon adenocarcinoma . IgA antibodies directed against mutated P38936 ras-D12 protein were detected in 51 ( 32 % ) of 160 colon cancer patients , but only in 1 ( 2.5 % ) of 40 normal individuals . The greater incidence of antibody in cancer patients provides presumptive evidence that immunization to the ras proteins occurred as a result of the malignancy . Examination of sera for antibody reactivity to wild-type P38936 ras protein ( denoted P38936 ras-G12 ) as well as P38936 ras proteins bearing the D12 , V12 , P28222 , or L61 mutations showed that antibody detected was largely to normal segments of the P38936 ras protein . Epitope mapping , using peptide neutralization assays with mutated or normal ras peptides as competitors , demonstrated that in 10 ( 67 % ) of 15 sera examined the antibody reactivity to P38936 ras-G12 protein was neutralized by peptides near the carboxyl terminus of P38936 ras protein , but not by peptides spanning the specific point mutation region . Antibody reactivities correlated with peripheral blood lymphocyte count , but did not correlate with patient age , sex , histology , stage , tumor locus , lymph node metastasis , or serum carcinoembryonic antigen . Characterisation of the contractile activity of eletriptan at the canine vascular P28222 receptor . The functional activity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] - 1 H-indole ) at the contractile serotonin ( 5-hydroxytryptamine ; 5-HT ) ' 1B-like ' receptor in dog isolated saphenous vein and basilar artery was investigated . DB00216 , like 5-HT and sumatriptan potently contracted saphenous vein ( pEC50 : 6.3 , 6.9 and 6.1 , respectively ) and basilar artery ( pEC50 7.2 , 7.5 and 6.8 , respectively ) . The maximum responses evoked by eletriptan was , unlike sumatriptan , significantly lower than that to 5-HT ( intrinsic activity saphenous vein : eletriptan 0.57 , 5-HT 1.0 , sumatriptan 0.85 ; basilar artery : eletriptan 0.77 , 5-HT 0.98 , sumatriptan 0.89 ) . Contractions evoked by eletriptan were antagonised by the P28222 /1D receptor antagonist GR125743 ( N- [ 4-methoxy-3- ( 4-methyl piperazin-1-yl ) phenyl ] -3-methyl-4-(4-pyridyl)benzamide ) with pA2 values of 9.1 in saphenous vein and 9.4 in basilar artery . Affinity estimates ( pKA ) for 5-HT and sumatriptan determined from receptor alkylation studies in saphenous vein were 6.6 and 6.3 , respectively , compared to the apparent equilibrium dissociation constant ( pKp ) for eletriptan of 6.8 . The rank order of relative intrinsic efficacies ( epsilon ) was 5-HT > sumatriptan > eletriptan . Thus , eletriptan required greater receptor occupancy ( 4.4-fold ) to evoke an equivalent contraction to 5-HT and sumatriptan in dog isolated saphenous vein . These data demonstrate that eletriptan is a potent partial agonist at the canine vascular P28222 receptor . DB00216 Pfizer . Pfizer has developed and launched eletriptan , a P28222 /1D agonist , for the potential treatment of migraine with and without aura . DB00216 has 6-fold greater affinity for the P28221 receptor than sumatriptan , and a 3-fold greater affinity for the P28222 receptor [ 249570 ] . DB00216 pharmacology has also been evaluated in vitro in comparison with zolmitriptan ( AstraZeneca plc ) and naratriptan ( GlaxoSmithKline plc ) [ 290116 ] . Plasma levels of DB02527 , cGMP and P80511 in sildenafil-induced headache . DB00203 , a selective inhibitor of the cyclic guanosine monophosphate ( cGMP ) degrading phosphodiestrase 5 ( O76074 ) , induced migraine without aura in 10 of 12 migraine patients and in healthy subjects it induced significantly more headache than placebo . The aim of the present study was to determine whether the pain-inducing effects of sildenafil would be reflected in plasma levels of important signalling molecules in migraine : cGMP , cyclic adenosine monophosphate ( DB02527 ) and calcitonin gene-related peptide ( P80511 ) . Ten healthy subjects ( four women , six men ) and 12 patients ( 12 women ) suffering from migraine without aura were included in two separate double-blind , placebo-controlled , cross-over studies in which placebo or sildenafil 100 mg was administered orally . Plasma levels of P80511 , DB02527 and cGMP were determined in blood from the antecubital vein . Despite the ability of sildenafil to induce headache and migraine , no significant differences in plasma levels of P80511 , cGMP and DB02527 were detected after sildenafil compared with placebo . In conclusion , plasma levels of P80511 , cGMP and DB02527 remain normal during sildenafil-induced headache or migraine . However , since previous studies indicate an important role of these signalling molecules , the present study questions whether DB02527 and cGMP in peripheral blood can be used for monitoring pathophysiological events in headache and migraine mechanisms . The effect of a single intraperitoneal dose of hrIL-1 alpha on DB05875 - , neurokinin A- , calcitonin gene-related peptide- and neuropeptide Y-like immunoreactivity in cerebrospinal fluid , plasma and knee joint synovial fluid in the rat . Substance P ( SP ) - , neurokinin A ( P20366 ) - , calcitonin gene-related peptide ( P80511 ) - and neuropeptide Y ( P01303 ) -like immunoreactivity ( -LI ) was studied in rats ' cerebrospinal fluid ( P04141 ) , plasma and synovial fluid ( SF ) from both knee joints at 2 and 24 h following an intraperitoneal administration of 0.05 ml human recombinant interleukin-1 alpha ( i.p. hrIL-1 alpha ) or saline . Increased or decreased levels of SP- , P20366 and P80511 -LI were detected in P04141 and plasma , whereas P01303 -LI was unaffected . In SF only P80511 -LI increased bilaterally . There was a correlation in P80511 -LI content between plasma and P04141 following i.p . hrIL-1 alpha but not between plasma and SF or P04141 and SF . It can be concluded that ( 1 ) i.p . hrIL-1 alpha activates somatosensory afferents thereby increasing SP- and P80511 -LI content in P04141 and plasma and P20366 -LI in P04141 ; 2 ) i.p . hrIL-1 alpha induces a bilateral increase of P80511 -LI in SF which is not mediated through systemic circulation and is possibly a part of the general host defensive reaction . Serotonergic stimulation of corticotropin-releasing hormone and pro-opiomelanocortin gene expression . The neurotransmitter serotonin ( 5-HT ) stimulates adrenocorticotropic hormone ( DB01285 ) secretion from the anterior pituitary gland via activation of central 5-HT1 and 5-HT2 receptors . The effect of 5-HT is predominantly indirect and may be mediated via release of hypothalamic corticotropin-releasing hormone ( P06850 ) . We therefore investigated the possible involvement of P06850 in the serotonergic stimulation of DB01285 secretion in male rats . Increased neuronal 5-HT content induced by systemic administration of the precursor 5-hydroxytryptophan ( 5-HTP ) in combination with the 5-HT reuptake inhibitor fluoxetine raised P06850 mRNA expression in the paraventricular nucleus ( PVN ) by 64 % , increased pro-opiomelanocortin ( P01189 ) mRNA in the anterior pituitary lobe by 17 % and stimulated DB01285 secretion five-fold . Central administration of 5-HT agonists specific to P08908 , P28222 , 5- Q13049 or P28335 receptors increased P06850 mRNA in the PVN by 15-50 % , P01189 mRNA in the anterior pituitary by 15-27 % and DB01285 secretion three- to five-fold , whereas a specific 5- Q9H205 agonist had no effect . Systemic administration of a specific anti- P06850 antiserum inhibited the DB01285 response to 5-HTP and fluoxetine and prevented the 5-HTP and fluoxetine-induced P01189 mRNA response in the anterior pituitary lobe . Central or systemic infusion of 5-HT increased DB01285 secretion seven- and eight-fold , respectively . Systemic pretreatment with the anti- P06850 antiserum reduced the DB01285 responses to 5-HT by 80 % and 64 % , respectively . It is concluded that 5-HT via activation of P08908 , 5- Q13049 , P28335 and possibly also P28222 receptors increases the synthesis of P06850 in the PVN and P01189 in the anterior pituitary lobe , which results in increased DB01285 secretion . Furthermore , the results indicate that P06850 is an important mediator of the DB01285 response to 5-HT . Effects of eletriptan on the peptidergic innervation of the cerebral dura mater and trigeminal ganglion , and on the expression of c-fos and c-jun in the trigeminal complex of the rat in an experimental migraine model . Nociceptive axons and terminals in the supratentorial cerebral dura mater display an intense calcitonin gene-related peptide ( P80511 ) immunoreactivity . In an experimental migraine model , it has been shown that electrical stimulation of the rat trigeminal ganglion induced an increase in the lengths of P80511 -immunoreactive axons , increased size and number of pleomorphic axonal varicosities in the dura mater , and an increased number of c-jun and c-fos protein-expressing nerve cells in the trigeminal complex . We demonstrate the effect of the highly specific and moderately lipophilic serotonin agonist eletriptan ( Pfizer ) which prevents the effects of electrical stimulation in the dura mater . DB00216 also affected the caudal trigeminal complex ; it markedly reduced the numbers of the oncoprotein-expressing cells , mainly after stimulation and to some extent also in nonstimulated animals . DB00216 also affected expression of P80511 in perikarya of trigeminal ganglion cells , insofar as the number of small nerve cells exhibiting a compact P80511 immunoreaction was decreased to one quarter of the original value . In all these respects , eletriptan acted in a similar way to sumatriptan , with the notable exception that eletriptan also blocked the stimulation-induced effects in the nucleus caudalis trigemini and the upper cervical spinal cord ( trigeminal complex ) , whereas sumatriptan did not . It is concluded that eletriptan , acting on perikarya and both the peripheral and the central axon terminals of primary sensory neurons , exerts its antimigraine effect by an agonist action on P28222 /1D receptors throughout the entire trigeminal system , probably by passing the blood-brain-barrier because of its lipophilic character . P40189 -linked signal transduction promotes the differentiation and maturation of dendritic cells . In order to explore the role of P40189 -linked signal transduction in the differentiation and maturation of dendritic cells ( DC ) , the mAb , B- P28222 , an agonist of P40189 , was used for the activation of P40189 on DC . The effects of cytokines and of anti- P40189 mAb on the proliferation of DC , and their expression of IL-12 and P33681 ( P33681 -1 ) by DC were evaluated . DC differentiating from peripheral blood mononuclear cells did not express the P05231 receptor alpha chain , but expressed P40189 . Anti- P40189 mAb promoted the proliferation of DC , induced by P05112 and granulocyte macrophage colony stimulating factor ( GM- P04141 ) , by up-regulating the GM- P04141 receptor on DC . DC induced by P40189 mAb and cytokines expressed DC-derived CC chemokine , as measured by RT-PCR . Induced DC also stimulated strong proliferation of autologous T cells in mixed lymphocyte reaction since an up-regulated expression of IL-12 and P33681 ( P33681 -1 ) was observed in DC activated by anti- P40189 mAb . Thus , P40189 signal transduction is important for the differentiation and maturation of DC .

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Rosiglitazone regulates P05231 -stimulated lipolysis in porcine adipocytes . Interleukin ( IL ) -6 , a proinflammatory cytokine , stimulates adipocyte lipolysis and induces insulin resistance in obese and diabetic subjects . However , the effects of the anti-diabetic drug rosiglitazone on P05231 -stimulated lipolysis and the underlying molecular mechanism are largely unknown . In this study , we demonstrated that rosiglitazone suppressed P05231 -stimulated lipolysis in differentiated porcine adipocytes by inactivation of extracellular signal-related kinase ( P29323 ) . Meanwhile , rosiglitazone enhanced the lipolysis response of adipocytes to isoprenaline . In addition , rosiglitazone significantly reversed P05231 -induced down-regulation of several genes such as perilipin A , peroxisome proliferators activated receptor gamma ( Q07869 & gamma ; ) , and fatty acid synthetase , as well as the up-regulation of P05231 mRNA . However , mRNA expression of Q07869 & gamma ; coactivator-1 alpha ( DB01053 -1 & alpha ; ) was enhanced by rosiglitazone in P05231 -stimulated adipocytes . These results indicate that rosiglitazone suppresses P05231 -stimulated lipolysis in porcine adipocytes through multiple molecular mechanisms . Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor . Mouse pluripotent hematopoietic stem cells ( PHSC ) were fractionated based on size and density using counterflow centrifugal elutriation ( CCE ) . These heterogeneous PHSC populations were further enriched by subtraction of cells with lineage-specific markers ( Lin- ) followed by positive sorting for c-kit expression . The cells were characterized for their functional and biochemical properties . We defined a subpopulation of c-kit-positive cells that expressed high numbers of c-kit receptors ( c-kitBR ) . One hundred c-kitBR cells from either low- or higher-density fractions were sufficient to repopulate the lymphohematopoietic system in WBB6F1-W/Wv ( W/Wv ) recipients , whereas no PHSC were found in cells with low ( c-kitDULL ) or no ( c-kitNEG ) c-kit expression . Lin- c-kitBR cells were separated into RhoDULL and RhoBR subsets based on their ability to efflux rhodamine 123 ( Rho ) . The PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin- c-kitBR RhoBR cells correlated directly with the number of day 12 colony-forming unit-spleen ( CFU- P28222 ) in each fraction . We were not able to enrich further for PHSC using monoclonal antibodies to the cell-surface markers AA4.1 or P01730 , which have been used by others to isolate PHSC . The small , low-density Lin- c-kitBR subset contained PHSC and few CFU- P28222 . This enabled us to assay PHSC for expression of the flk-2 gene , which encodes a tyrosine kinase receptor present on fetal liver PHSC . Purified RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA . We suggest that AA4.1 , P01730 and flk-2 are expressed as stage-specific markers on PHSC in cell cycle . Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . DB01095 inhibits growth and alters the malignant phenotype of the P13671 glioma cell line . BACKGROUND : DB01095 is a member of the family of P04035 inhibitors ( statins ) extensively used in medical practice . Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development . The aim of the present study was to investigate the anti-cancer potential of fluvastatin in P13671 rat malignant glioma cells . METHODS : First , the effects of fluvastatin on cell viability ( MTT assay ) , proliferation ( BrdU assay ) , cell morphology , and cytoskeleton were examined . Subsequently , its effect on extracellular signal regulated kinase 1 and 2 ( P27361 /2 ) and P45983 and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 and P15692 was determined using a wound-healing assay and ELISA test , respectively . RESULTS : The results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor P13671 cells ( IC(50) = 8.6 μM , 48 h ) , but did not inhibit the growth of normal neuronal cells . The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm , chromatin condensation , and nucleus breakdown . CONCLUSION : The inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p- P27361 /2 expression , upregulation of p- P45983 /2 , and reduction in the P14780 and P15692 concentrations in culture media . The high anticancer ( antiproliferative , proapoptotic , antiinvasive ) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma . P06850 and the blood-brain-barrier . Increased blood-brain-barrier ( BBB ) permeability precedes any clinical or pathologic signs and is critical in the pathogenesis of multiple sclerosis ( MS ) and brain metastases . P01730 + Q8IXH7 cells mediate demyelination in MS , but how they get sensitized and enter the brain to induce brain inflammation remains obscure . TH2 cytokines associated with allergic disorders have recently been implicated in MS , while genes upregulated in MS plaques include the mast cell-specific tryptase , the IgE receptor ( Fc-epsilon-RI ) and the histamine-1 receptor . Mast cell specific tryptase is elevated in the P04141 of MS patients , induces microvascular leakage and stimulates protease-activated receptors ( PAR ) , leading to widespread inflammation . BBB permeability , MS and brain metastases appear to worsen in response to acute stress that leads to the local release of corticotropin-releasing hormone ( P06850 ) , which activates brain mast cells to selectively release P05231 , P10145 and vascular endothelial growth factor ( P15692 ) . Acute stress increases BBB permeability that is dependent on P06850 and mast cells . Acute stress shortens the time of onset of experimental alleric encephalomyelitis ( EAE ) that does not develop in W/W mast cell deficient or P06850 -/- mice . Brain mast cell inhibition and P34998 antagonists offer novel therapeutic possibilities . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . Differential gene expression in relation to the clinical characteristics of human brain arteriovenous malformations . Arteriovenous malformations ( AVMs ) of the central nervous system are considered as congenital disorders . They are composed of abnormally developed dilated arteries and veins and are characterized microscopically by the absence of a capillary network . We previously reported DNA fragmentation and increased expression of apoptosis-related factors in AVM lesions . In this article , we used microarray analysis to examine differential gene expression in relation to clinical manifestations in 11 AVM samples from Japanese patients . We categorized the genes with altered expression into four groups : death-related , neuron-related , inflammation-related , and other . The death-related differentially expressed genes were P14780 , P15018 , P04179 , Q16548 , P39900 , and P17066 . The neuron-related genes were P01303 , P06702 , Q15784 , S100Abeta , Q9UQM7 , Q8TBG9 , P08172 , and Q8NCB2 . The inflammation-related genes were PTX3 , P10145 , P05231 , P02778 , P32455 , P20309 , P09341 , P27930 , P55774 , and Q99616 . In addition , we compared gene expression in those with or without clinical characteristics including deep drainer , embolization , and high-flow nidus . We identified a small number of genes . Using these microarray data we are able to generate and test new hypotheses to explore AVM pathophysiology . Microarray analysis is a useful technique to study clinical specimens from patients with brain vascular malformations . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . Ras signaling mechanisms underlying impaired GluR1-dependent plasticity associated with fragile X syndrome . Fragile X syndrome , caused by the loss of Q06787 gene function and loss of fragile X mental retardation protein ( Q06787 ) , is the most commonly inherited form of mental retardation . The syndrome is characterized by associative learning deficits , reduced risk of cancer , dendritic spine dysmorphogenesis , and facial dysmorphism . However , the molecular mechanism that links loss of function of Q06787 to the learning disability remains unclear . Here , we report an examination of small GTPase Ras signaling and synaptic AMPA receptor ( AMPA-R ) trafficking in cultured slices and intact brains of wild-type and Q06787 knock-out mice . In Q06787 knock-out mice , synaptic delivery of GluR1- , but not GluR2L- and P48058 -containing AMPA-Rs is impaired , resulting in a selective loss of GluR1-dependent long-term synaptic potentiation ( LTP ) . Although Ras activity is upregulated , its downstream MEK ( extracellular signal-regulated kinase kinase ) - P29323 ( extracellular signal-regulated kinase ) signaling appears normal , and phosphoinositide 3-kinase ( PI3K ) -protein kinase B ( P31749 ; or Akt ) signaling is compromised in Q06787 knock-out mice . Enhancing Ras-PI3K- P31749 signaling restores synaptic delivery of GluR1-containing AMPA-Rs and normal LTP in Q06787 knock-out mice . These results suggest aberrant Ras signaling as a novel mechanism for fragile X syndrome and indicate manipulating Ras-PI3K- P31749 signaling to be a potentially effective approach for treating patients with fragile X syndrome . Partial agonists of the α3β4* neuronal nicotinic acetylcholine receptor reduce ethanol consumption and seeking in rats . DB00898 use disorders ( AUDs ) impact millions of individuals and there remain few effective treatment strategies . Despite evidence that neuronal nicotinic acetylcholine receptors ( nAChRs ) have a role in AUDs , it has not been established which subtypes of the nAChR are involved . Recent human genetic association studies have implicated the gene cluster P32297 - P30532 - P30926 encoding the α3 , α5 , and β4 subunits of the nAChR in susceptibility to develop nicotine and alcohol dependence ; however , their role in ethanol-mediated behaviors is unknown due to the lack of suitable and selective research tools . To determine the role of the α3 , and β4 subunits of the nAChR in ethanol self-administration , we developed and characterized high-affinity partial agonists at α3β4 nAChRs , CP-601932 , and PF-4575180 . Both CP-601932 and PF-4575180 selectively decrease ethanol but not sucrose consumption and operant self-administration following long-term exposure . We show that the functional potencies of CP-601932 and PF-4575180 at α3β4 nAChRs correlate with their unbound rat brain concentrations , suggesting that the effects on ethanol self-administration are mediated via interaction with α3β4 nAChRs . Also varenicline , an approved smoking cessation aid previously shown to decrease ethanol consumption and seeking in rats and mice , reduces ethanol intake at unbound brain concentrations that allow functional interactions with α3β4 nAChRs . Furthermore , the selective α4β2(*) nAChR antagonist , DHβE , did not reduce ethanol intake . Together , these data provide further support for the human genetic association studies , implicating P32297 and P30926 genes in ethanol-mediated behaviors . CP-601932 has been shown to be safe in humans and may represent a potential novel treatment for AUDs . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Statin decreases endothelial microparticle release from human coronary artery endothelial cells : implication for the Rho-kinase pathway . OBJECTIVE : Elevated plasma levels of endothelial microparticles ( EMPs ) are associated with the presence of clinical atherosclerosis . Considering the anti-inflammatory properties of P04035 inhibitors on the endothelium , we studied the effect of fluvastatin on the release of EMPs in cultured human coronary artery endothelial cells ( HCAEC ) . METHODS AND RESULTS : EMPs were generated in P01375 -activated HCAECs . The absolute number of EMPs was enumerated using a novel two-color flow cytometric immunostaining technique with TruCount beads as an internal reference . EMPs are defined as EC membrane vesicles ( 1-2 microm in size ) with a characteristic immunophenotype . The addition of fluvastatin to P01375 -activated HCAECs significantly suppressed Q7L5Y9 release . DB01095 suppressed P01375 -induced Rho activation . The Rho-kinase inhibitor , Y-27632 , reproduced the effect of statin . CONCLUSION : Q7L5Y9 release from P01375 -activated HCAECs is suppressed by fluvastatin . In addition , the Rho/Rho-kinase may play an important role in modulating Q7L5Y9 release . Serious obstetric complications interact with hypoxia-regulated/vascular-expression genes to influence schizophrenia risk . The etiology of schizophrenia is thought to include both epistasis and gene-environment interactions . We sought to test whether a set of schizophrenia candidate genes regulated by hypoxia or involved in vascular function in the brain ( P31749 , P23560 , O75052 , P36544 , P21964 , Q96EV8 , Q99259 , Q14832 , Q99466 , Q02297 , O43272 , P49798 , P01375 ) interacted with serious obstetric complications to influence risk for schizophrenia . A family-based study of transmission disequilibrium was conducted in 116 trios . Twenty-nine probands had at least one serious obstetric complication ( OC ) using the McNeil-Sjostrom Scale , and many of the OCs reported were associated with the potential for fetal hypoxia . Analyses were conducted using conditional logistic regression and a likelihood ratio test ( LRT ) between nested models was performed to assess significance . Of the 13 genes examined , four ( P31749 ( three SNPs ) , P23560 ( two SNPs ) , Q96EV8 ( one SNP ) and Q14832 ( one SNP ) ) showed significant evidence for gene-by-environment interaction ( LRT P-values ranged from 0.011 to 0.037 ) . Although our sample size was modest and the power to detect interactions was limited , we report significant evidence for genes involved in neurovascular function or regulated by hypoxia interacting with the presence of serious obstetric complications to increase risk for schizophrenia . [ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type-1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 +/- 9.1 years , duration of diabetes 18 +/- 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 +/- 13 vs 140 +/- 13 mm Hg , P < 0.05 ; diastolic pressures 89 +/- 10 vs 87 +/- 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g/24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg/mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 +/- 14.4 vs 8.8 +/- 8.1 U/g creatinine ; P < 0.01 ) . DB01197 did not affect metabolic control ( HbA1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy . Genetics and alcoholism . DB00898 is widely consumed ; however , excessive use creates serious physical , psychological and social problems and contributes to the pathogenesis of many diseases . DB00898 use disorders ( that is , alcohol dependence and alcohol abuse ) are maladaptive patterns of excessive drinking that lead to serious problems . Abundant evidence indicates that alcohol dependence ( alcoholism ) is a complex genetic disease , with variations in a large number of genes affecting a person 's risk of alcoholism . Some of these genes have been identified , including two genes involved in the metabolism of alcohol ( P00325 and P05091 ) that have the strongest known affects on the risk of alcoholism . Studies continue to reveal other genes in which variants affect the risk of alcoholism or related traits , including P47869 , P08172 , P48051 and Q8WXX7 . As more variants are analysed and studies are combined for meta-analysis to achieve increased sample sizes , an improved picture of the many genes and pathways that affect the risk of alcoholism will be possible . Hypoxic damage to the periventricular white matter in neonatal brain : role of vascular endothelial growth factor , nitric oxide and excitotoxicity . The present study examined factors that may be involved in the development of hypoxic periventricular white matter damage in the neonatal brain . Wistar rats ( 1-day old ) were subjected to hypoxia and the periventricular white matter ( corpus callosum ) was examined for the mRNA and protein expression of hypoxia-inducible factor-1alpha ( HIF-1alpha ) , endothelial , neuronal and inducible nitric oxide synthase ( P29474 , P29475 and P35228 ) , vascular endothelial growth factor ( P15692 ) and N-methyl-D-aspartate receptor subunit 1 ( Q05586 ) between 3 h and 14 days after hypoxic exposure by real-time RT-PCR , western blotting and immunohistochemistry . Up-regulated mRNA and protein expression of HIF-1alpha , P15692 , Q05586 , P29474 , P29475 and P35228 in corpus callosum was observed in response to hypoxia . Q05586 and P35228 expression was found in the activated microglial cells , whereas P15692 was localized to astrocytes . An enzyme immunoassay showed that the P15692 concentration in corpus callosum was significantly higher up to 7 days after hypoxic exposure . NO levels , measured by colorimetric assay , were also significantly higher in hypoxic rats up to 14 days after hypoxic exposure as compared with the controls . A large number of axons undergoing degeneration were observed between 3 h and 7 days after the hypoxic exposure at electron-microscopic level . Our findings point towards the involvement of excitotoxicity , P15692 and NO in periventricular white matter damage in response to hypoxia . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . The execution of the transcriptional axis mutant p53 , Q01094 and P47928 promotes tumor neo-angiogenesis . P47928 ( inhibitor of DNA binding 4 ) is a member of a family of proteins that function as dominant-negative regulators of basic helix-loop-helix transcription factors . Growing evidence links ID proteins to cell proliferation , differentiation and tumorigenesis . Here we identify P47928 as a transcriptional target of gain-of-function p53 mutants R175H , R273H and R280K . Depletion of mutant p53 protein severely impairs P47928 expression in proliferating tumor cells . The protein complex mutant p53- Q01094 assembles on specific regions of the P47928 promoter and positively controls P47928 expression . The P47928 protein binds to and stabilizes mRNAs encoding pro-angiogenic factors P10145 and P09341 -alpha . This results in the increase of the angiogenic potential of cancer cells expressing mutant p53 . These findings highlight the transcriptional axis mutant p53 , Q01094 and P47928 as a still undefined molecular mechanism contributing to tumor neo-angiogenesis . DB00898 blocks leukocyte recruitment and endothelial cell activation in vivo and in vitro . Immune system impairment and increased susceptibility to infection among alcohol abusers is a significant but not well-understood problem . We hypothesized that acute ethanol administration would inhibit leukocyte recruitment and endothelial cell activation during inflammation and infection . Using LPS and carrageenan air pouch models in mice , we found that physiological concentrations of ethanol ( 1-5 g/kg ) significantly blocked leukocyte recruitment ( 50-90 % ) . Because endothelial cell activation and immune cell-endothelial cell interactions are critical regulators of leukocyte recruitment , we analyzed the effect of acute ethanol exposure on endothelial cell activation in vivo using the localized Shwartzman reaction model . In this model , ethanol markedly suppressed leukocyte accumulation and endothelial cell adhesion molecule expression in a dose-dependent manner . Finally , we examined the direct effects of ethanol on endothelial cell activation and leukocyte-endothelial cell interactions in vitro . DB00898 , at concentrations within the range found in human blood after acute exposure and below the levels that induce cytotoxicity ( 0.1-0.5 % ) , did not induce endothelial cell activation , but significantly inhibited P01375 -mediated endothelial cell activation , as measured by adhesion molecule ( P16581 , P05362 , P19320 ) expression and chemokine ( P10145 , P13500 , RANTES ) production and leukocyte adhesion in vitro . Studies exploring the potential mechanism by which ethanol suppresses endothelial cell activation revealed that ethanol blocked NF-kappaB nuclear entry in an P25963 -dependent manner . These findings support the hypothesis that acute ethanol overexposure may increase the risk of infection and inhibit the host inflammatory response , in part , by blocking endothelial cell activation and subsequent immune cell-endothelial cell interactions required for efficient immune cell recruitment . Downstream effects of striatal-enriched protein tyrosine phosphatase reduction on RNA expression in vivo and in vitro . Striatal-enriched protein tyrosine phosphatase ( P54829 ) is a brain-specific tyrosine phosphatase that has been shown to de-phosphorylate several key neuronal signaling proteins , including kinases ( extracellular signal-regulated kinase ( P27361 /2 ) , P06241 , Q14289 ) and glutamate receptor subunits ( N-methyl-d-aspartate receptor subtype 2B ( Q13224 ) , glutamate receptor 2 ( P42262 ) ) . Step knock-out mice have increased phosphorylation of these substrates in the brain , with potential functional consequences in synaptic plasticity and cognitive tasks . It is therefore of interest to identify the molecular pathways and downstream transcriptional targets that are impacted by Step knockdown . In the present study , striatal RNA samples from Step wild-type , knock-out and heterozygous mice were hybridized to Affymetrix microarray chips and evaluated for transcriptional changes between genotypes . Pathway analysis highlighted Erk signaling and multiple pathways related to neurotrophin signaling , neuronal development and synaptic transmission . Potential genes of interest identified by microarray were confirmed by quantitative real-time polymerase chain reaction ( qRT-PCR ) in the cortex and hippocampus , which shared several transcriptional alterations with the striatum . In order to evaluate Step knockdown in an in vitro system , a panel of genes were evaluated using qRT-PCR in rat cortical neurons that were transduced with lentivirus expressing short hairpin RNA against Step or a non-targeting control . Our data suggest that Step has a role in the expression of immediate early genes relevant to synaptic plasticity , in both in vitro and in vivo systems . New findings on the genetic influences on alcohol use and dependence . PURPOSE OF REVIEW : DB00898 dependence is a complex disorder with a well documented highly hereditary nature . This article reviews the recent advances in our understanding of the direct and indirect genetic influences on alcohol use and dependence . RECENT FINDINGS : Recent findings can be summarized as follows : ( a ) twin studies have defined and estimated the risks of general and specific alcohol-related vulnerabilities . ( b ) Linkage studies have provided largely inconsistent findings , though several chromosomal regions have been implicated . ( c ) Quantitative trait loci analyses in animals have identified that the Mpdz gene predisposes to alcohol dependence and withdrawal . ( d ) Examination of family-based samples has identified several genes including P47869 and P08172 thought to be associated with alcohol dependence . SUMMARY : Despite great advances in understanding of genetic vulnerability in alcohol use disorders , only two gene complexes , DB00067 and P05091 , have been identified as having defined effects on alcohol use and liability to dependence in humans . New genes associated with increased risks for the disorder will certainly be added to this list in the near future . Neurobiological analyses of the effects of these genes will surely contribute to further understanding of the cause of alcohol dependence and the interindividual differences in risks . DB01296 improves cardiac function following trauma-hemorrhage by increased protein O-GlcNAcylation and attenuation of NF-{kappa}B signaling . We have previously demonstrated that in a rat model of trauma-hemorrhage ( T-H ) , glucosamine administration during resuscitation improved cardiac function , reduced circulating levels of inflammatory cytokines , and increased tissue levels of O-linked N-acetylglucosamine ( O-GlcNAc ) on proteins . The mechanism(s) by which glucosamine mediated its protective effect were not determined ; therefore , the goal of this study was to test the hypothesis that glucosamine treatment attenuated the activation of the nuclear factor-kappaB ( NF-kappaB ) signaling pathway in the heart via an increase in protein O-GlcNAc levels . Fasted male rats were subjected to T-H by bleeding to a mean arterial blood pressure of 40 mmHg for 90 min followed by resuscitation . DB01296 treatment during resuscitation significantly attenuated the T-H-induced increase in cardiac levels of P01375 and P05231 mRNA , IkappaB-alpha phosphorylation , NF-kappaB , NF-kappaB DNA binding activity , P05362 , and P05164 activity . LPS ( 2 microg/ml ) increased the levels of IkappaB-alpha phosphorylation , P01375 , P05362 , and NF-kappaB in primary cultured cardiomyocytes , which was significantly attenuated by glucosamine treatment and overexpression of O-GlcNAc transferase ; both interventions also significantly increased O-GlcNAc levels . In contrast , the transfection of neonatal rat ventricular myocytes with O15294 small-interfering RNA decreased O-GlcNAc transferase and O-GlcNAc levels and enhanced the LPS-induced increase in IkappaB-alpha phosphorylation . DB01296 treatment of macrophage cell line RAW 264.7 also increased O-GlcNAc levels and attenuated the LPS-induced activation of NF-kappaB . These results demonstrate that the modulation of O-GlcNAc levels alters the response of cardiomyocytes to the activation of the NF-kappaB pathway , which may contribute to the glucosamine-mediated improvement in cardiac function following hemorrhagic shock . Protein kinase Czeta mediates micro-opioid receptor-induced cross-desensitization of chemokine receptor P51681 . We have previously shown that the μ-opioid receptor ( MOR ) is capable of mediating cross-desensitization of several chemokine receptors including P51681 , but the biochemical mechanism of this process has not been fully elucidated . We have carried out a series of functional and biochemical studies and found that the mechanism of MOR-induced cross-desensitization of P51681 involves the activation of PKCζ . Inhibition of PKCζ by its pseudosubstrate inhibitor , or its siRNA , or dominant negative mutants suppresses the cross-desensitization of P51681 . Our results further indicate that the activation of PKCζ is mediated through a pathway involving phosphoinositol-dependent kinase-1 ( PDK1 ) . In addition , activation of MOR elevates the phosphorylation level and kinase activity of PKCζ . The phosphorylation of PKCζ can be suppressed by a dominant negative mutant of PDK1 . We observed that following MOR activation , the interaction between PKCζ and PDK1 is immediately increased based on the analysis of fluorescent resonance energy transfer in cells with the expression of PKCζ-YFP and PDK1- P27918 . In addition , cells expressing PKCζ kinase motif mutants ( Lys-281 , DB00156 -410 , DB00156 -560 ) fail to exhibit full MOR-induced desensitization of P51681 activity . Taken together , we propose that upon DAMGO treatment , MOR activates PKCζ through a PDK1-dependent signaling pathway to induce P51681 phosphorylation and desensitization . Because P51681 is a highly proinflammatory receptor , and a critical coreceptor for HIV-1 , these results may provide a novel approach for the development of specific therapeutic agents to treat patients with certain inflammatory diseases or AIDS . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . No evidence for association between 19 cholinergic genes and bipolar disorder . Cholinergic dysfunction has been proposed for the pathogenesis of bipolar disorder ( BD ) , and we have therefore performed a systematic association study of cholinergic system genes in BD ( including schizoaffective disorder bipolar type ) . We genotyped 93 single nucleotide polymorphisms ( SNPs ) in 19 genes ( P28329 , P11229 -5 , P02708 -7 , Q9UGM1 , Q9GZZ6 , and P11230 -4 ) in two series of samples : the National Institute of Mental Health ( NIMH ) Genetics Initiative pedigrees with 474 samples from 152 families , and the Clinical Neurogenetics ( CNG ) pedigrees with 83 samples from 22 multiplex families . Sib-transmission/disequilibrium test ( sib_TDT ) analysis showed nominally significant transmission bias for four SNPs ( Q15822 : rs7017417 , P = 0.024 ; P30532 : rs514743 , P = 0.031 ; P11230 : rs2302762 , P = 0.049 ; P30926 : rs1948 , P = 0.031 ) . Haploview analyses showed nominally significant transmission bias of several haplotypes in Q15822 , P36544 , P11230 , and P30926 , respectively . However , none of these associations reached gene-wide significance after correction by permutation . DB00898 dependence ( including alcohol abuse ) was not a significant covariate in the present genetic association analysis . Thus , it is unlikely that these 19 cholinergic genes play a major role in the pre-disposition to BD in these pedigrees . Predicting the effect of naltrexone and acamprosate in alcohol-dependent patients using genetic indicators . DB00659 and naltrexone are effective medications in the treatment of alcoholism . However , effect sizes are modest . Pharmacogenomics may improve patient-treatment-matching and effect sizes . It is hypothesized that naltrexone exerts its effect through genetic characteristics associated with the dopaminergic/opioidergic positive reinforcement system , whereas acamprosate works through the glutamatergic/GABAergic negative reinforcement system . DB00898 -dependent subjects were randomly assigned to either acamprosate or naltrexone . Subjects participated in a cue-exposure experiment at the day before and at the last day of medication . Reductions in cue-induced craving and physiological cue reactivity were measured . Differential effects of naltrexone and acamprosate on these outcomes were tested for different polymorphisms of the opioid , dopamine , glutamate and GABA-receptors . Significant matching effects were found for polymorphisms at the P14416 , Q16445 and P47870 gene . In addition , a trend was found for the P35372 polymorphism . This provides evidence for the matching potential of genotypes . It is expected that more effective treatments can be offered when genetic information is used in patient-treatment-matching . DB00898 increases desensitization of recombinant P48058 AMPA receptor and TARP combinations . Glutamate receptors are important target molecules of the acute effect of ethanol . We studied ethanol sensitivity of homomeric P48058 receptors expressed in human embryonic kidney 293 cells and examined whether recently discovered transmembrane alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ( AMPA ) receptor regulatory proteins ( TARPs ) affect ethanol sensitivity . Coexpression of the TARPs , stargazin , and gamma4 increased the time constant ( tau-value ) of current decay in the presence of agonist , thus slowing the onset of desensitization and increasing the steady-state current . DB00898 produced less inhibition of the peak current than the steady-state current for all types of the P48058 receptors . In addition , ethanol concentration-dependently accelerated the rate of desensitization , measured as the tau-value of fast decay of peak current . This effect was enhanced with coexpression of TARPs . The recovery from desensitization was slowed down by coexpression of gamma4 but ethanol did not affect this process in any P48058 combination . The results support the idea that increased desensitization is an important mechanism in the ethanol inhibition of AMPA receptors and indicate that coexpression of TARPs can alter this effect of ethanol . Selective increases of AMPA , DB01221 , and kainate receptor subunit mRNAs in the hippocampus and orbitofrontal cortex but not in prefrontal cortex of human alcoholics . Glutamate is the main excitatory transmitter in the human brain . Drugs that affect the glutamatergic signaling will alter neuronal excitability . DB00898 inhibits glutamate receptors . We examined the expression level of glutamate receptor subunit mRNAs in human post-mortem samples from alcoholics and compared the results to brain samples from control subjects . RNA from hippocampal dentate gyrus ( HP-DG ) , orbitofrontal cortex ( OFC ) , and dorso-lateral prefrontal cortex ( DL- P27918 ) samples from 21 controls and 19 individuals with chronic alcohol dependence were included in the study . Total RNA was assayed using quantitative RT-PCR . Out of the 16 glutamate receptor subunits , mRNAs encoding two AMPA [ 2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid ] receptor subunits P42262 and P42263 ; three kainate receptor subunits Q13002 , Q13003 and Q16478 and five DB01221 ( N-methyl-D-aspartate ) receptor subunits Q05586 , Q12879 , Q14957 , O15399 , and Q8TCU5 were significantly increased in the HP-DG region in alcoholics . In the OFC , mRNA encoding the DB01221 receptor subunit Q8TCU5 was increased , whereas in the DL- P27918 , no differences in mRNA levels were observed . Our laboratory has previously shown that the expression of genes encoding inhibitory GABA-A receptors is altered in the HP-DG and OFC of alcoholics ( Jin et al. , 2011 ) . Whether the changes in one neurotransmitter system drives changes in the other or if they change independently is currently not known . The results demonstrate that excessive long-term alcohol consumption is associated with altered expression of genes encoding glutamate receptors in a brain region-specific manner . It is an intriguing possibility that genetic predisposition to alcoholism may contribute to these gene expression changes . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . DB01197 attenuates matrix metalloproteinase-2 and -9 in monocrotaline-induced right ventricular hypertrophy in rats . Little is known about the influence of angiotensin converting enzyme ( P12821 ) inhibitors on matrix metalloproteinase ( MMP ) in right ventricular remodeling . We investigated the effect of captopril , an P12821 inhibitor , on P08253 and P14780 in monocrotaline-induced right ventricular hypertrophy . Six-week-old male Wistar rats were injected intraperitoneally with monocrotaline ( 60 mg/kg ) or saline . The rats were administrated captopril ( 30 mg/kg per day ) or a vehicle orally for 24 days from the day of monocrotaline injection . At day 25 , echocardiography was performed and hearts were excised . Expressions and activities of P08253 and P14780 were measured by Western blotting and by gelatin zymography , respectively . In monocrotaline-injected rats , right ventricular weight/tail length ratio increased significantly . Histological analysis revealed cardiomyocyte hypertrophy and fibrosis in right ventricular sections . Echocardiography showed right ventricular dysfunction compared with saline-injected rats . The right ventricular hypertrophy , fibrosis , and dysfunction were inhibited by captopril . However , captopril did not attenuate an increase in pulmonary artery pressure . P08253 and P14780 expressions and activities in right ventricles increased significantly in monocrotaline-injected rats and captopril inhibited them . These findings indicate that captopril attenuates the development of monocrotaline-induced right ventricular hypertrophy in association with inhibition of P08253 and P14780 in rats . Platelet activation and arterial peripheral serotonin turnover in cardiac remodeling associated to aortic stenosis . Peripheral serotonin ( 5-HT ) has been involved in adverse cardiac remodeling and valve fibrosis . The peripheral levels of 5-HT mainly depend on its release from activated platelets and degradation by monoamine oxidase A ( P21397 ) . The SERAOPI study investigated the relationship between arterial serotoninergic system , degree of platelet activation and cardiac remodeling , in patients with aortic valve stenosis ( AS ) . Thirty patients with severe AS and 15 control subjects underwent transthoracic echocardiography , radial , and aortic arterial blood sampling . Measurements of 5-HT and its P21397 -dependent degradation product , 5-HIAA , were performed by HPLC . Arterial platelet activation was assessed by flow cytometry analysis of platelet surface expression of P16109 and activated integrin P08514 /IIIa . Activated platelets and arterial plasma 5-HT increased in AS patients as compared to control subjects ( P16109 1.08 ± 0.2MFI vs. 0.49 ± 0.1MFI , P = 0.04 ; P08514 /IIIa 0.71 ± 0.1MFI vs. 0.35 ± 0.1MFI ; P = 0.0015 and arterial plasma 5-HT 11.55 ± 1.6 nM vs. 6.18 ± 0.7 nM , P = 0.028 , respectively ) . Moreover , 5-HT was strongly correlated to left ventricular hypertrophy assessed by echocardiography . The correlation was independent of cardiovascular risk comorbidities and others echocardiographic AS parameters . Finally , plasma 5-HIAA increased in AS patients ( 74.64 ± 9.7 nM vs. 37.16 ± 4.1 nM ; P = 0.0002 ) indicating a higher 5-HT degradation rate by P21397 . Platelet activation , arterial circulating serotonin , and serotonin degradation increased in patients with AS . These observations suggest that the serotoninergic system may contribute to the pathogenesis of AS including valve fibrosis and adverse ventricular remodeling . Emergence of motor circuit activity . In the developing nervous system , ordered neuronal activity patterns can occur even in the absence of sensory input and to investigate how these arise , we have used the model system of the embryonic chicken spinal motor circuit , focusing on motor neurons of the lateral motor column ( O15467 ) . At the earliest stages of their molecular differentiation , we can detect differences between medial and lateral O15467 neurons in terms of expression of neurotransmitter receptor subunits , including P30532 , P36544 , Q12879 , P39086 , P08908 and P28222 , as well as the Q9H2X9 transporter . Using patch-clamp recordings we also demonstrate that medial and lateral O15467 motor neurons have subtly different activity patterns that reflect the differential expression of neurotransmitter receptor subunits . Using a combination of patch-clamp recordings in single neurons and calcium-imaging of motor neuron populations , we demonstrate that inhibition of nicotinic , muscarinic or GABA-ergic activity , has profound effects of motor circuit activity during the initial stages of neuromuscular junction formation . Finally , by analysing the activity of large populations of motor neurons at different developmental stages , we show that the asynchronous , disordered neuronal activity that occurs at early stages of circuit formation develops into organised , synchronous activity evident at the stage of O15467 neuron muscle innervation . In light of the considerable diversity of neurotransmitter receptor expression , activity patterns in the O15467 are surprisingly similar between neuronal types , however the emergence of patterned activity , in conjunction with the differential expression of transmitter systems likely leads to the development of near-mature patterns of locomotor activity by perinatal ages . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . Association of Q05940 gene polymorphisms with alcohol dependence . DB00898 -related diseases cause significant harm in the western world . Up to 65 % of the phenotypic variance is genetically determined . Few candidate genes have been identified , comprising P08319 , P05091 , P21964 , P34998 , Q01959 ( Q01959 ) , P47869 and P21397 . While abnormalities in the dopaminergic mesolimbic reward system are considered important mediators of alcoholism , studies analyzing variants of dopamine receptors showed conflicting results . Other modulators of the reward system are synaptosomal genes . Among candidate genes , polygenic variants of the Vesicular Monamine Transporter 2 ( Q05940 ) gene locus associated with alterations of drinking behavior were published . These variants comprise single nucleotide polymorphisms ( SNPs ) within the promoter region and the open reading frame . In this study , we confirm the association of Q05940 SNP rs363387 ( allelic association : p = 0.015 ) with alcohol dependence . This SNP defines several haplotypes including up to four SNPs ( minimal p = 0.0045 ) . In addition , numeric effects in the subgroups of males and patients with positive family history were found . We suggest that several rs363387 T-allele containing haplotypes increase the risk of alcohol dependence ( OR 1.53 ) , whereas G-allele containing haplotypes confer protection against alcohol dependence . Taken together , there is supporting evidence for a contribution of Q05940 gene variants to phenotypes of alcohol dependence . An in vitro model of human acute ethanol exposure that incorporates P49682 - and P61073 -dependent recruitment of immune cells . Alcoholic liver disease ( P33897 ) is one of the commonest causes of cirrhosis and liver failure in the developed world . Hepatic inflammation is the critical stage in progression of both P33897 and non- P33897 , but it remains difficult to study the underlying mechanisms in a human system , and current animal models do not fully recapitulate human liver disease . We developed a human tissue-based system to study lymphocyte recruitment in response to ethanol challenge . Precision-cut liver slices ( PCLS ) from human livers were incubated in culture , and hepatic function was determined by albumin production , 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide assay , glucose uptake responses , and morphometric assessment . Responses of tissue and lymphocytes to ethanol exposure were determined by PCR , flow cytometry , histology , and lymphocyte infiltration assays . Human PCLS demonstrated appropriate upregulation of P05181 , ADH1α , and P00326 in response to ethanol treatment . DB00898 also induced expression of endothelial P19320 and P05362 , production of sICAM-1 and P10145 , and the chemokine receptors P49682 and P61073 on P01730 and CD8 lymphocytes . P49682 - and P61073 -dependent migration of lymphocytes into the tissue increased significantly in response to treatment with ethanol . We have demonstrated that ethanol increases chemokine receptor expression and lymphocyte recruitment into human liver tissue , suggesting that it may operate directly to promote hepatitis in P33897 . The physiological and pathophysiological responses of the PCLS to ethanol in vitro highlight the potential of this assay for dissecting the molecular mechanisms underlying human liver inflammation and as a screening tool for novel therapeutics . P12821 inhibitors could be therapeutic for antisocial personality disorder . Antisocial personality traits are an important topic for research . The societal cost of these behaviors encourages efforts at a better understanding of central nervous system causes . Catecholamine genes are being studied to facilitate this understanding , and some tentative findings are being reached about several of these genes . It seems that many genes play a role to produce antisocial behaviors so complexity of elucidating each gene is obvious . One conclusion that could be drawn from the current research findings is that DA2 like receptors ( P14416 , P35462 , P21917 ) with alleles that decrease neurotransmission are facilitatory of antisocial behaviors . DA2 like receptors cause neuronal firing to inhibit many peripheral functions through adenylyl cyclase inhibition . When these receptors are less active by genetically decreased density , lower affinity , or by low dopamine levels as final common pathways then inhibition is released and a state of disinhibition can be said to describe this state . Peripheral metabolism is increased and behavioral activation is noted . P00797 is disinhibited in this setting thus allowing sympathetic nervous system activation . The fight or flight behaviors thus produced , in the extreme , would be the setting of antisocial behavior . Research validates this hypothesis . Understanding this final common pathway toward antisocial behavior should lead to better treatment for individuals with this pattern of behavior before they have caused harm to themselves and others . P12821 inhibitors are well tolerated drugs used in the treatment of hypertension and heart failure and would also treat antisocial behavior disorders . A genomic insight into diversity among tribal and nontribal population groups of Manipur , India . Twenty autosomal markers , including linked markers at two gene markers , are used to understand the genomic similarity and diversity among three tribal ( Paite , Thadou , and Kom ) and one nontribal communities of Manipur ( Northeast India ) . Two of the markers ( P01730 and HB9 ) are monomorphic in Paite and one ( the P01730 marker ) in Kom . Data suggest the Meitei ( nontribal groups ) stand apart from the three tribal groups with respect to higher heterozygosity ( 0.366 ) and presence of the highest ancestor haplotypes of P14416 markers ( 0.228 ) ; this is also supported by principal co-ordinate analysis . These populations are found to be genomically closer to the Chinese population than to other Indian populations . Genetic variation in the P30532 gene affects mRNA levels and is associated with risk for alcohol dependence . DB00898 dependence frequently co-occurs with cigarette smoking , another common addictive behavior . Evidence from genetic studies demonstrates that alcohol dependence and smoking cluster in families and have shared genetic vulnerability . Recently a candidate gene study in nicotine dependent cases and nondependent smoking controls reported strong associations between a missense mutation ( rs16969968 ) in exon 5 of the P30532 gene and a variant in the 3'-UTR of the P32297 gene and nicotine dependence . In this study we performed a comprehensive association analysis of the P30532 - P32297 - P30926 gene cluster in the Collaborative Study on the Genetics of Alcoholism ( COGA ) families to investigate the role of genetic variants in risk for alcohol dependence . Using the family-based association test , we observed that a different group of polymorphisms , spanning P30532 - P32297 , demonstrate association with alcohol dependence defined by Diagnostic and Statistical Manual of Mental Disorders , 4th edn ( DSM-IV ) criteria . Using logistic regression we replicated this finding in an independent case-control series from the family study of cocaine dependence . These variants show low linkage disequilibrium with the SNPs previously reported to be associated with nicotine dependence and therefore represent an independent observation . Functional studies in human brain reveal that the variants associated with alcohol dependence are also associated with altered steady-state levels of P30532 mRNA . Loss of the candidate tumor suppressor Q14201 triggers acute cellular senescence via the P29323 - O15054 -p16(INK4a) signaling axis . The B-cell translocation gene 3 ( Q14201 ) is a member of the antiproliferative BTG gene family and a downstream target of p53 . Q14201 also binds and inhibits Q01094 . Although it connects functionally two major growth-regulatory pathways , the physiological role of Q14201 remains largely uncharacterized . Here , we present evidence that loss of Q14201 in normal cells induced cellular senescence , which was correlated with enhanced P29323 - P05412 signaling and elevated expression of the histone H3K27me3 demethylase O15054 / O15054 , leading to acute induction of p16(INK4a) . Importantly , we also found that Q14201 expression is specifically downregulated in prostate cancer , thus providing a physiological link with human cancers . Our data suggest that Q14201 may have a fail-safe role against tumorigenic progression . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . P30532 gene D398N polymorphism in Japanese lung adenocarcinoma . BACKGROUND : Recently , to identify genetic factors that modify lung cancer risk , P30532 non-synonymous variant amino acid position 398 ( D398N ) was identified . The site was a highly conserved in the second cellular loop of the nicotinic acetylcholine receptor subunit protein . MATERIALS AND METHODS : We have investigated P30532 gene polymorphism status in 302 surgically treated lung adenocarcinoma cases from Nagoya City University Hospital . The presence or absence of P30532 polymorphism was analyzed by direct sequences . P00533 mutations status was already investigated and reported . RESULTS : We detected nine cases ( 2.98 % ) of P30532 polymorphism ( D398N ) in our cohort . Total P00533 mutations were present in 129 patients ( 42.7 % ) . The polymorphism statuses were not correlated with gender ( women ; 2.1 % versus men ; 3.7 % , P = 0.5119 ) , smoking status ( never smoker ; 2.0 % versus smoker ; 4.0 % , P = 0.3339 ) , pathological stages ( stage I ; 2.6 % versus stage II-IV ; 3.8 % , P = 0.7246 ) , and P00533 mutation status of the lung adenocarcinomas ( mutation ; 2.3 % versus wild type ; 3.7 % , P = 0.7373 ) . In this analysis , P30532 polymorphism ( D398N ) patients had significantly worse prognosis ( 5/9 were dead ; mean survival = 27.1 mo ) than the patients with P30532 wild type ( 74/293 were dead ; mean survival = 113.9 mo ) ( log-rank test ; P = 0.0146 ) . CONCLUSION : Although P30532 polymorphism is rare from Japanese lung cancer , polymorphism status might be correlated with shorter survival . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . DB00898 -related expectancies are associated with the D2 dopamine receptor and GABAA receptor beta3 subunit genes . Molecular genetic research has identified promising markers of alcohol dependence , including alleles of the D2 dopamine receptor ( P14416 ) and the GABAA receptor beta3 subunit ( P28472 ) genes . Whether such genetic risk manifests itself in stronger alcohol-related outcome expectancies , or in difficulty resisting alcohol , is unknown . In the present study , A1+ ( A1A1 and A1A2 genotypes ) and A1- ( A2A2 genotype ) alleles of the P14416 and P55008 + ( G1G1 and P55008 non- P55008 genotypes ) and P55008 - ( non- P55008 non- P55008 genotype ) alleles of the P28472 gene were determined in a group of 56 medically ill patients diagnosed with alcohol dependence . Mood-related alcohol expectancy ( AE ) and drinking refusal self-efficacy ( DRSE ) were assessed using the Drinking Expectancy Profile ( Manual for the Drinking Expectancy Profile , Behaviour Research and Therapy Centre , Brisbane , 1996 ) . Patients with the P14416 A1+ allele , compared with those with the P14416 A1- allele , reported significantly lower DRSE in situations of social pressure . Similarly , lower DRSE was reported under social pressure by patients with the P28472 P55008 + allele when compared to those with the P28472 P55008 - alleles . Patients with the P28472 P55008 + allele also revealed reduced DRSE in situations characterized by negative affect than those with the P28472 P55008 - alleles . Patients carrying the P28472 P55008 + allele showed stronger AE relating to negative affective change ( for example , increased depression ) than their P28472 P55008 - counterparts . Biological influence in the development of some classes of cognitions is hypothesized . The clinical implications , particularly with regard to patient-treatment matching and the development of an integrated psychological and pharmacogenetic approach , are discussed . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . DB00898 dose-dependently elicits opposing regulatory effects on hippocampal AMPA receptor P42262 subunits through a zeta inhibitory peptide-sensitive kinase in adolescent and adult Sprague-Dawley rats . AMPA receptor P42262 subunits are strongly implicated in cognition , and prior work suggests that these subunits may be regulated by atypical protein kinase C ( aPKC ) isoforms . The present study assessed whether hippocampal and cortical AMPA receptor P42262 subunit regulation may be an underlying factor in known age-related differences to cognitive-impairing doses of ethanol , and if aPKC isoforms modulate such responses . Hippocampal AMPA receptor P42262 subunit , protein kinase Mζ ( PKMζ ) , and PKCι/λ expression were elevated during adolescence compared to adults . 1 h following a low-dose ( 1.0-g/kg ) ethanol exposure , hippocampal AMPA receptor P42262 subunit serine 880 phosphorylation was decreased in adolescents , but was increased in adults . Age-dependent changes in P42262 subunit phosphorylation were paralleled by alterations in aPKC isoforms , and zeta inhibitory peptide ( Q8N5A5 ) administration prevented ethanol-induced increases in both in adults . DB00898 -induced changes in P42262 subunit phosphorylation were associated with delayed regulation in synaptosomal P42262 subunit expression 24 h later . A higher ethanol dose ( 3.5-g/kg ) failed to elicit changes in most measures in the hippocampus at either age . Similar to the hippocampus , analysis of cerebral cortical tissue also revealed age-related declines . However , no demonstrable effects were found following a low-dose ethanol exposure at either age . High-dose ethanol exposure reduced adolescent P42262 subunit phosphorylation and aPKC isoform expression that were again accompanied by delayed reductions in synaptosomal P42262 subunit expression . Together , these results suggest that P42262 -containing AMPA receptor modulation by aPKC isoforms is age- , region- and dose-dependently regulated , and may potentially be involved in developmentally regulated ethanol-induced cognitive impairment and other ethanol behaviors . Therapy with interferon-beta modulates endogenous catecholamines in lymphocytes of patients with multiple sclerosis . OBJECTIVE : To investigate the endogenous dopaminergic/adrenergic system of lymphocytes in multiple sclerosis ( MS ) patients during treatment with interferon ( IFN ) -beta . METHODS : Patients with relapsing-remitting MS undergoing IFN-beta treatment were prospectively studied during the first year of treatment . Circulating lymphocytes were obtained at baseline and after 1 , 3 , 6 and 12 months of treatment and assayed for catecholamine ( CA ) production and mRNA expression of tyrosine hydroxylase ( TH , the rate-limiting enzyme in the synthesis of CA ) , beta(2)-adrenoceptors ( AR ) and D2 , D3 and D5 dopaminergic receptors ( DR ) . RESULTS : In cells from patients treated with IFN-beta for 12 months the production of CA hugely increased and was less sensitive to P01579 -induced inhibition . Expression of mRNA for TH , beta(2)-AR and P21918 was already enhanced after 1 month and further increased up to 6-12 months of treatment . On the contrary , P14416 mRNA progressively decreased and P35462 mRNA did not significantly change over the whole study period . CONCLUSIONS : In MS patients IFN-beta treatment enhances the ability of lymphocytes to produce CA , and induces extensive modifications of both beta(2)-AR and DR-operated pathways . The clinical relevance of these effects deserves consideration . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Cdk5 phosphorylates dopamine D2 receptor and attenuates downstream signaling . The dopamine D2 receptor ( P14416 ) is a key receptor that mediates dopamine-associated brain functions such as mood , reward , and emotion . P12004 -dependent kinase 5 ( Cdk5 ) is a proline-directed serine/threonine kinase whose function has been implicated in the brain reward circuit . In this study , we revealed that the serine 321 residue ( S321 ) in the third intracellular loop of P14416 ( D2i3 ) is a novel regulatory site of Cdk5 . Cdk5-dependent phosphorylation of S321 in the D2i3 was observed in in vitro and cell culture systems . We further observed that the phosphorylation of S321 impaired the agonist-stimulated surface expression of P14416 and decreased G protein coupling to P14416 . Moreover , the downstream DB02527 pathway was affected in the heterologous system and in primary neuronal cultures from p35 knockout embryos likely due to the reduced inhibitory activity of P14416 . These results indicate that Cdk5-mediated phosphorylation of S321 inhibits P14416 function , providing a novel regulatory mechanism for dopamine signaling .

MH_train_1054

MH_train_1054

MH_train_1054

interacts_with DB00790?

multiple_choice

Promoter polymorphism of cyclooxygenase-1 ( P23219 ) is not associated with ulcerative colitis in the Japanese population . BACKGROUND/AIMS : Non-steroidal anti-inflammatory drugs ( NSAIDs ) aggravate or ameliorate clinical disease activity in patients with ulcerative colitis ( UC ) . UC is associated with increased local production of prostanoids . This study attempted to clarify the relationship between cyclooxygenase-1 ( P23219 ) gene polymorphisms ( T-1676C and A-842G/C50T ) and ulcerative colitis in a Japanese population . METHODOLOGY : The study was performed on 108 patients with UC ( mean age : Q04695 years , M:F=56:52 ) and 293 healthy controls ( mean age : 49.0 years , M:F=161:132 ) . The PCR-SSCP method was employed to detect P23219 polymorphisms using DNA extracted from peripheral blood cells . RESULTS : No A-842G/C50T polymorphism was detected in all 401 Japanese subjects . The frequency of -1676C allele of P23219 gene in HC group and UC group was 48.8 % and 48.1 % , respectively . T-1676C genotypes of P23219 did not show significant association with UC risk . In addition , these genotypes were not also associated with the clinicopathological parameters of UC . CONCLUSIONS : This research suggests that P23219 promoter polymorphisms ( T-1676C and A-842G/ C50T ) may not be associated with the risk of developing ulcerative colitis in a Japanese population . P35354 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways . The functional significance of protease-activated receptors ( PARs ) in endothelial cells is largely undefined , and the intracellular consequences of their activation are poorly understood . Here , we show that the serine protease thrombin , a P25116 -selective peptide ( TFLLRN ) , and SLIGKV ( P55085 -selective peptide ) induce cyclooxygenase-2 ( P35354 ) protein and mRNA expression in human endothelial cells without modifying P23219 expression . P35354 induction was accompanied by sustained production of 6-keto-PGF1alpha , the stable hydrolysis product of prostacyclin , and this was inhibited by indomethacin and the P35354 -selective inhibitor NS398 . P25116 and P55085 stimulation rapidly activated both P27361 /2 and p38MAPK , and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha formation . Thrombin and peptide agonists of P25116 and P55085 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus , and this , as well as PAR-induced 6-keto-PGF1alpha synthesis , was inhibited by co-infection with adenovirus encoding wild-type or mutated ( Y42F ) P25963 . Thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132 . Activation of P25116 or P55085 promoted nuclear translocation and phosphorylation of p65-NF-kappaB , and thrombin-induced but not P55085 -induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK . Activation of Q96RI0 by AYPGKF increased phosphorylation of P27361 /2 and p38MAPK without modifying NF-kappaB activation or P35354 induction . Our data show that P25116 and P55085 , but not Q96RI0 , are coupled with P35354 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on P27361 /2 , p38MAPK , and P25963 -dependent NF-kappaB activation . Consequences of the Y139F Vkorc1 mutation on resistance to AVKs : in-vivo investigation in a 7th generation of congenic Y139F strain of rats . OBJECTIVES : In humans , warfarin is used as an anticoagulant to reduce the risk of thromboembolic clinical events . DB00682 derivatives are also used as rodenticides in pest control . The gene encoding the protein targeted by anticoagulants is the Vitamin K-2,3-epoxide reductase subunit 1 ( Q9BQB6 ) . Since its discovery in 2004 , various amino acid and transcription-regulatory altering Q9BQB6 mutations have been identified in patients who required extreme antivitamin K dosages , or wild populations of rodents that were difficult to control with anticoagulant rodenticides . One unresolved question concerns the dependency of the Q9BQB6 on the genetic background in humans and rodents that respond weakly or not at all to anticoagulants . Moreover , an important question requiring further analyses concerns the role of the Vkorc1 gene in mediating resistance to more recently developed warfarin derivatives ( superwarfarins ) . METHODS : In this study , we bred a quasicongenic rat strain by using a wild-caught anticoagulant resistant rat as a donor to introduce the Y > F amino acid change at position 139 in the Vkorc1 into the genetic background of an anticoagulant susceptible Spraque-Dawley recipient strain . RESULTS AND CONCLUSION : In this manuscript we report the prothrombin times measured in the P08709 generation after exposure to chlorophacinone , bromadiolone , difenacoum and difethialone . We observed that the mutation Y139F mediates resistance in an otherwise susceptible genetic background when exposed to chlorophacinone and bromadiolone . However , the physiological response to the super-warfarins , difenacoum and difethialone , may be strongly dependent on other genes located outside the congenic interval ( 28.3 cM ) bracketing the Vkorc1 in our P08709 generation congenic strain . Cyclic nucleotide-gated channels , calmodulin , adenylyl cyclase , and calcium/calmodulin-dependent protein kinase II are required for late , but not early , long-term memory formation in the honeybee . Memory is a dynamic process that allows encoding , storage , and retrieval of information acquired through individual experience . In the honeybee Apis mellifera , olfactory conditioning of the proboscis extension response ( O15534 ) has shown that besides short-term memory ( P0DMM9 ) and mid-term memory ( A1L3X4 ) , two phases of long-term memory ( LTM ) are formed upon multiple-trial conditioning : an early phase ( e-LTM ) which depends on translation from already available mRNA , and a late phase ( l-LTM ) which requires de novo transcription and translation . Here we combined olfactory O15534 conditioning and neuropharmacological inhibition and studied the involvement of the NO-cGMP pathway , and of specific molecules , such as cyclic nucleotide-gated channels ( CNG ) , calmodulin ( P62158 ) , adenylyl cyclase ( AC ) , and Ca(2+)/calmodulin-dependent protein kinase ( CaMKII ) , in the formation of olfactory LTM in bees . We show that in addition to NO-cGMP and DB02527 -PKA , CNG channels , P62158 , AC , and CaMKII also participate in the formation of a l-LTM ( 72-h post-conditioning ) that is specific for the learned odor . Importantly , the same molecules are dispensable for olfactory learning and for the formation of both A1L3X4 ( in the minute and hour range ) and e-LTM ( 24-h post-conditioning ) , thus suggesting that the signaling pathways leading to l-LTM or e-LTM involve different molecular actors . P02649 phenotypes and plasma lipids in young and middle-aged subjects . The relationship between apolipoprotein ( apo ) E phenotypes and the levels of plasma lipid , lipoprotein and apo E in young ( mean , 21 years of age ) and middle-aged ( mean , 49 years of age ) subjects was investigated . Apo E phenotypes were determined by a rapid flat gel isoelectric focusing method that we had developed previously . Young subjects with apo E3/2 and E4/3 had significantly higher levels of plasma triglyceride ( TG ) , very low density lipoprotein ( VLDL ) -TG and VLDL-cholesterol than those with apo E3/3 . Middle-aged subjects with apo E3/2 ( 54.5 % ) and E4/3 ( Q04695 % ) had higher frequency of hyperlipoproteinemia ( mainly type IV ) than those with apo E3/3 ( 25.8 % ) . Furthermore , the middle-aged subjects with apo E3/2 had significantly higher levels of plasma TG , VLDL-TG and apo E , and significantly lower levels of plasma high density lipoprotein-cholesterol than those with apo E3/3 . These results indicate that apo E phenotype E3/2 and E4/3 are associated with lipid abnormalities even in young subjects , which may be caused by impaired functions of apo E2 and E4 . Bayesian meta-analysis of tissue angiotensin-converting enzyme inhibitors for reduction of adverse cardiovascular events in patients with diabetes mellitus and preserved left ventricular function . The role of angiotensin-converting enzyme ( P12821 ) inhibitors in diabetic patients with preserved ventricular function is uncertain . Tissue P12821 inhibitors have been defined by increased lipophilicity and structural characteristics that result in greater tissue-specific P12821 binding when compared with plasma P12821 inhibitors . A Bayesian meta-analysis of randomized trials was conducted to evaluate tissue P12821 inhibitors in prevention of cardiovascular disease among patients with diabetes mellitus and preserved left ventricular function . Four trials were selected that evaluated 2 different P12821 inhibitors and included 10,328 patients ( 43,517 patient-years ) . The DB00790 Substudy in Coronary Artery Disease and Diabetes ( PERSUADE ) and the DB00790 Protection Against Recurrent Stroke Study ( PROGRESS ) compared the effects of perindopril vs a placebo , and the Heart Outcomes Prevention Evaluation ( HOPE ) and the Non- P01308 -Dependent Diabetes , Hypertension , Microalbuminuria , Proteinuria , Cardiovascular Events , and Ramipril ( DIABHYCAR ) study investigated the impact of ramipril vs a placebo . Bayesian meta-analysis of sequential trials and sensitivity analysis of therapeutic response were subsequently computed . Bayesian meta-analysis determined reduced risk of cardiovascular mortality ( PB=.991 ) , myocardial infarction ( PB=.999 ) , and the need for invasive coronary revascularization ( PB=.995 ) when compared with placebo . Total mortality was also decreased ( PB=.967 ) , while the risk of stroke ( PB=.907 ) and hospitalization for heart failure ( PB=.923 ) were impacted . Bayesian meta-analysis of randomized trials suggests that tissue P12821 inhibitors decrease the probability that diabetic patients with preserved left ventricular function will experience myocardial infarctions and cardiovascular death and reduce overall mortality . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Association of sixty-one non-synonymous polymorphisms in forty-one hypertension candidate genes with blood pressure variation and hypertension . We previously selected a group of hypertension candidate genes by a key word search using the OMIM database of NCBI and validated 525 coding single nucleotide polymorphisms ( SNPs ) in 179 hypertension candidate genes by DNA sequencing in a Japanese population . In the present study , we examined the association between 61 non-synonymous SNPs and blood pressure variations and hypertension . We used DNA samples taken from 1,880 subjects in the Suita study , a population-based study using randomly selected subjects . Analyses of covariance adjusting for age , body mass index , hyperlipidemia , diabetes , smoking , drinking , and antihypertensive medication revealed that 17 polymorphisms in 16 genes ( P04114 , CAST , P51801 , O60931 , P10912 , P13807 , P08603 , O95163 , Q14654 , P11150 , P06858 , P41231 , Q15165 , P02730 , TRH , P04275 ) were significantly associated with blood pressure variations . Multivariate logistic regression analysis with adjustment for the same factors revealed that 11 polymorphisms in 11 genes ( CAST , P16410 , P12259 , GC , P10912 , P11150 , Q13093 , P02730 , SLCI8A1 , TRH , P04275 ) showed significant associations with hypertension . Five polymorphisms in five genes , CAST(calpastatin) , P11150 ( hepatic lipase ) , P02730 ( band 3 anion transporter ) , TRH ( thyrotropin-releasing hormone ) , and P04275 ( P04275 ) , were significantly associated with both blood pressure variation and hypertension . Thus , our study suggests that these five genes were susceptibility genes for essential hypertension in this Japanese population . Dual effect of angiotensin-converting enzyme inhibition on angiogenesis in type 1 diabetic mice . OBJECTIVE : We analyzed the beneficial therapeutic effect of angiotensin converting enzyme inhibitor ( ACEI ) on both retinal and hind limb neovascularization in diabetic mice . METHODS AND RESULTS : Diabetic mice ( streptozotocin , 40 mg/kg ) were treated with or without ACEI ( DB00790 , 3 mg/kg per day ) or AT1 receptor blocker ( DB00796 , 20 mg/kg ) for 4 months . Hind limb ischemia was then induced by right femoral artery ligature for 1 additional month . In the ischemic leg , angiographic score , capillary density , and foot perfusion were increased by 2.7 , 2.0-fold , and 1.6-fold , respectively , in ACEI-treated diabetic mice compared with untreated diabetic animals ( P < 0.01 ) . ACEI also raised vascular endothelial growth factor ( P15692 ) protein level by 1.4-fold in ischemic diabetic leg . This ACEI pro-angiogenic effect was totally blunted in diabetic bradykinin B2 receptor-deficient animals , suggesting that it was mediated by the bradykinin pathway . In the diabetic retina , angiotensinogen and P12821 mRNA levels were increased by 2.8-fold and 4.1-fold , respectively ( P < 0.01 versus nondiabetic mice ) , highlighting a local activation of renin-angiotensin system . Diabetes also raised P15692 protein level by 1.5-fold ( P < 0.05 versus nondiabetic mice ) . Treatments with ACEI and AT1 receptor blocker hampered diabetes-induced P15692 upregulation and retinal neovascularization . CONCLUSIONS : P12821 inhibition improved neovascularization in the diabetic ischemic leg through activation of bradykinin signaling , whereas it reduced vessel growth in the diabetic retina through inhibition of overacting Ang II pathway . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . A role for plasma transforming growth factor-beta and matrix metalloproteinases in aortic aneurysm surveillance in Marfan syndrome ? BACKGROUND : We have previously shown that the angiotensin-converting enzyme ( P12821 ) inhibitor perindopril reduced aortic diameter by 3-7mm in Marfan syndrome ( MFS ) patients . Excessive signalling by the transforming growth factor-beta ( TGF-beta ) has been implicated in the development of aortic dilatation . We hypothesised that reduction in aortic diameter would correlate with reduction in plasma TGF-beta and matrix metalloproteinase ( MMP ) levels . METHODS : 17 MFS patients ( aged 33+/-5 ( mean+/-SD ) ) on standard beta-blocker therapy were randomised to also receive perindopril ( n=10 ) or placebo ( n=7 ) for 24 weeks in a double blind study . Aortic root diameters were assessed at four sites via transthoracic echocardiography . Venous blood samples were analysed for latent and active TGF-beta , P08253 and P08254 levels . RESULTS : DB00790 significantly reduced aortic root diameters relative to placebo in both end-systole and end-diastole ( by 1.2-3mm/m(2) , p < 0.001 ) . In addition , compared to placebo perindopril significantly reduced latent TGF-beta levels by 14.0+/-4.5ng/ml ( p=0.01 ) , active TGF-beta levels by 4+/-1ng/ml ( p=0.02 ) , P08253 levels by 22+/-6ng/ml ( p < 0.001 ) , and P08254 levels by 5+/-1ng/ml ( p < 0.001 ) . There were moderately strong correlations between the pre/post intervention change in aortic diameters and the change in both latent ( r=0.49-0.76 , p=0.001-0.04 ) and active TGF-beta ( r=0.59-0.73 , p=0.002-0.02 ) , P08253 ( r=0.63-0.75 , p=0.001-0.007 ) , and P08254 plasma levels ( r=0.81-0.83 , p < 0.0001 ) . CONCLUSIONS : Plasma TGF-beta , P08253 and P08254 should be further explored in longitudinal trials as potential prognostic indicators of progression of aortic dilatation and response to therapy in MFS . DB00790 : possible use in cancer therapy . Since angiogenesis is essential for the growth of any solid tumor , emerging efforts are being made to develop antiangiogenic therapy . To date , however , no antiangiogenic agent has become widely available for the clinical setting . Angiotensin I-converting enzyme ( P12821 ) inhibitors are commonly used as antihypertensive agents and it has recently been suggested that they decrease the risk of cancer . Studies have found that an P12821 inhibitor , perindopril , is a potent inhibitor of experimental tumor development and angiogenesis at a clinically comparable dose . The potent angiogenic factor , vascular endothelial growth factor ( P15692 ) , is significantly suppressed by perindopril and also inhibits P15692 -induced tumor growth . In vitro studies showed that perindopril is not cytotoxic to either tumor cells or endothelial cells . Since perindopril is already in widespread clinical use without serious side effects , it may represent a potential new strategy for anticancer therapy . DB00790 decreases P wave dispersion in patients with stage 1 hypertension . INTRODUCTION : P12821 inhibitors prevent atrial fibrillation episodes by effective control of blood pressure and improving electrical and structural remodelling in the atria . Increased P wave dispersion ( P35670 ) is a non-invasive electrocardiographic marker for paroxysmal atrial fibrillation . The aim of the study was to evaluate the effect of perindopril treatment on P35670 in hypertensive patients . METHODS : Forty-eight hypertensive patients ( mean age 57.4+/-11.8 years , 18 men ) were included . Blood pressure values were determined and 12-lead electrocardiograms were recorded at the beginning and at the first week , first month , third month and sixth month of the perindopril treatment.The difference between maximum and minimum P wave durations was calculated as P35670 . RESULTS : PWDs were significantly shortened at the first , third and sixth months ( 41.7+/-8.8 ms , Q04695 +/-6.9 ms and 38.3+/-7.1 ms , respectively ) compared with baseline and first-week measurements ( 54.3+/-9.2 ms and 49.0+/-9.1 ms , respectively , p < 0.001 ) . Baseline P35670 was correlated with body mass index ( r=0.32 , p=0.026 ) , while P35670 at the sixth month of treatment was significantly correlated with left atrial volume index ( r=0.30 , p=0.042 ) . Multiple linear regression analysis revealed that P35670 at the sixth month was related to baseline P35670 ( p=0.001 ) . CONCLUSION : DB00790 treatment significantly reduced P35670 in hypertensive patients . [ New immunoserological tests and pathogenesis for rheumatoid arthritis ] . In 1987 , the P10323 ( American College of rheumatology ) guidelines for classification of RA were published and the JCR ( Japan College of rheumatology ) classification of early diagnosis for RA was established in 1994 . The determination of rheumatoid factor ( RF ) was included in both the above classification standards . However , Ig-M type of RF has problems in specificity and sensitivity . Recent progress in various RA studies has revealed the molecular mechanism of RA and new therapy and laboratory tests ( ie. CA-RF , P08254 ) have been developed . Thus , determination changes in various diagnostic markers such as P08254 , and inflammatory cytokines ( ie. IL-1beta , P60568 , P05231 , P10145 , Q14116 ) are important for treatment of RA patients . Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . Identification of insulin-stimulated phosphorylation sites on calmodulin . P01308 enhances calmodulin phosphorylation in vivo . To determine the insulin-sensitive phosphorylation sites , phosphocalmodulin was immunoprecipitated from Chinese hamster ovary cells expressing human insulin receptors ( CHO/IR ) . P62158 was constitutively phosphorylated on serine , threonine , and tyrosine residues , and insulin enhanced phosphate incorporation on serine and tyrosine residues . Phosphocalmodulin immunoprecipitated from control and insulin-treated CHO/IR cells , and calmodulin phosphorylated in vitro by the insulin receptor kinase and casein kinase II were resolved by two-dimensional phosphopeptide mapping . Several common phosphopeptides were detected . The phosphopeptides from the in vitro maps were eluted and phosphoamino acid analysis , manual sequencing , strong cation exchange chromatography , and additional proteolysis were performed . This strategy demonstrated that DB00135 -99 and DB00135 -138 were phosphorylated in vitro by the insulin receptor kinase and DB00156 -79 , DB00133 -81 , DB00133 -101 and DB00156 -117 were phosphorylated by casein kinase II . In vivo phosphorylation sites were identified by comigration of phosphopeptides on two-dimensional maps with phosphopeptides derived from calmodulin phosphorylated in vitro and by phosphoamino acid analysis . This approach revealed that DB00135 -99 and DB00135 -138 of calmodulin were phosphorylated in CHO/IR cells in response to insulin . Additional sites remain to be identified . The identification of the insulin-stimulated in vivo tyrosine phosphorylation sites should facilitate the elucidation of the physiological role of phosphocal-modulin . The effect of abdominal obesity on insulin sensitivity and serum lipid and cytokine concentrations in African women . OBJECTIVES : Studies have shown clear associations of abdominal obesity with lipid and glucose metabolism and cytokine levels in a number of different population groups . However , no such studies have been performed in an African population in which visceral adipose tissue levels have been shown to be lower than in European subjects . DESIGN AND PATIENTS : Cross-sectional analysis in 124 African women . MEASUREMENTS : Fasting serum samples were taken from all subjects and anthropometric measurements obtained . Blood levels of glucose , insulin , total cholesterol , high-density lipoprotein ( HDL ) and low-density lipoprotein ( LDL ) cholesterol , triglyceride , interleukin ( IL ) -6 , P10145 and Q14116 were measured . Subjects were separated into normal and abnormal glucose tolerant groups and into tertiles according to waist circumference ( WC ) . P01308 resistance was assessed using the homeostasis model assessment ( HOMA ) . RESULTS : Abnormal glucose-tolerant subjects had higher WC , glucose and HOMA levels than the normal glucose-tolerant group . Increased WC was associated with higher triglyceride , insulin and HOMA levels and lower HDL levels . Multiple regression analyses showed that WC associated positively with HOMA and serum triglyceride levels and negatively with HDL levels . Q14116 was a positive but weak determinant of the HOMA level and BMI correlated positively with serum P05231 concentrations . CONCLUSIONS : Although previous studies have shown that African subjects have a lower visceral adipose depot size than European subjects , abdominal obesity is still associated with insulin resistance and dyslipidaemia . The association between abdominal obesity and metabolic dysfunction within this population is not dependent upon P05231 , P10145 or Q14116 . First report of warfarin dose requirements in patients possessing the P11712 *12 allele . BACKGROUND : DB00682 is the most frequently prescribed anticoagulant in North America and Europe . It is administered as a racemate , but S-warfarin is principally responsible for its anticoagulant activity . Cytochrome P450 ( CYP ) 2C9 is the enzyme primarily responsible for the metabolism of S-warfarin . Numerous variant alleles of P11712 have been identified . The P11712 *12 ( rs9332239 ) allele harbors a P489S substitution in P11712 which has been shown to result in a 40 % decline in catalytic activity in vitro . CASES : Four Caucasian patients with a low mean weekly warfarin dose ( MWWD ) were genotyped for P11712 , Q9BQB6 and P02649 variant alleles . None of the four patients carried the common P11712 variant alleles ( *2 , *3 , *5 , *6 , *7 , *8 , *9 , *11 , *13 ) despite a relatively low MWWD ( 23.4±7.94 mg ) compared to 208 patients carrying the CYP29C9*1 genotype ( 32.2±12.65 mg ) . Given that P11712 *12 confers decreased in vitro activity to the enzyme , we investigated whether these patients carried this allele . All four patients were P11712 *12 CT heterozygotes . Individual comparisons with patients possessing the same Q9BQB6 and P02649 genotypes also demonstrated lower dose requirements in the patients that possessed P11712 *12 allele . CONCLUSIONS : There are no reports of the clinical impact of rs9332239 on P11712 substrates . This is the first report of patients with the rare P11712 *12 genotype and lower warfarin dose requirements . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . Aromatic-turmerone attenuates invasion and expression of P14780 and P35354 through inhibition of NF-κB activation in TPA-induced breast cancer cells . Recent evidence suggests that breast cancer is one of the most common forms of malignancy in females , and metastasis from the primary cancer site is the main cause of death . Aromatic (ar)-turmerone is present in Curcuma longa and is a common remedy and food . In the present study , we investigated the inhibitory effects of ar-turmerone on expression and enzymatic activity levels of 12-O-tetradecanoylphorbol-13-acetate ( TPA ) -induced matrix metalloproteinase ( MMP ) -9 and cyclooxygenaase-2 ( P35354 ) in breast cancer cells . Our data indicated that ar-turmerone treatment significantly inhibited enzymatic activity and expression of P14780 and P35354 at non-cytotoxic concentrations . However , the expression of tissue inhibitor of metalloproteinase ( P01033 ) -1 , P16035 , P08253 , and P23219 did not change upon ar-turmerone treatment . We found that ar-turmerone inhibited the activation of NF-κB , whereas it did not affect AP-1 activation . Moreover , The ChIP assay revealed that in vivo binding activities of NF-κB to the P14780 and P35354 promoter were significantly inhibited by ar-turmerone . Our data showed that ar-turmerone reduced the phosphorylation of PI3K/Akt and P27361 /2 signaling , whereas it did not affect phosphorylation of JNK or p38 MAPK . Thus , transfection of breast cancer cells with PI3K/Akt and P27361 /2 siRNAs significantly decreased TPA-induced P14780 and P35354 expression . These results suggest that ar-turmerone suppressed the TPA-induced up-regulation of P14780 and P35354 expression by blocking NF-κB , PI3K/Akt , and P27361 /2 signaling in human breast cancer cells . Furthermore , ar-turmerone significantly inhibited TPA-induced invasion , migration , and colony formation in human breast cancer cells . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Genetic polymorphisms for the study of multifactorial stroke . Single-gene disorders explain only a minority of stroke cases . Stroke represents a complex trait , which is usually assumed to be polygenic . On this topic , the role of a wide number of candidate genes has been investigated in stroke through association studies , with controversial results . Therefore , it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke . This review is an update and an extensive analysis of the more recent association studies conducted in stroke . We evaluated a number of studies on several candidate genes ( including P12259 , F2 , P02671 / P02675 / P02679 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / Q9UKI9 / P04054 / P08514 , P17301 , P07359 , P12821 , AGT , NOS3 , P02649 , P06858 , P27169 , Q08499 , P20292 , P42898 , Q99707 , and P35520 ) , providing a final panel of genes and molecular variants . We categorized this panel in relation to the degree of association with stroke , supported by the results of meta-analyses and case-control studies . Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses . Vascular endothelial growth factor expression and glomerular endothelial cell loss in the remnant kidney model . BACKGROUND : Vascular endothelial growth factor ( P15692 ) is constitutively expressed in the glomerulus where it may have a role in the maintenance of capillary endothelial cell integrity . The present study sought to examine changes in P15692 expression in a model of progressive renal disease and to assess the effects of angiotensin converting enzyme ( P12821 ) inhibition . METHODS : Subtotal nephrectomized ( STNx ) rats were randomly assigned to receive vehicle ( n=10 ) or the P12821 inhibitor perindopril ( 8 mg/l drinking water ) for 12 weeks duration ( n=10 ) . Sham-operated rats were used as controls ( n=10 ) . Glomerular capillary endothelial cell density was evaluated by immunostaining for the pan-endothelial cell marker Q06609 -1 and P15692 expression was assessed by quantitative in situ hybridization . RESULTS : In STNx rats glomerular capillary endothelial cell density was reduced to 19 % that of sham rats ( P < 0.01 ) with a concomitant reduction in glomerular P15692 expression , also to 19 % of sham rats ( P < 0.01 ) . DB00790 treatment was associated with normalization of both capillary endothelial cell density and glomerular P15692 mRNA . CONCLUSIONS : Reduction in glomerular P15692 expression is a feature of the renal pathology that follows subtotal nephrectomy . In the context of the known functions of this growth factor , these findings suggest that diminution in P15692 may contribute to the demonstrated loss of glomerular endothelium that develops in this model of progressive renal disease . Genomics and the prospects of existing and emerging therapeutics for cardiovascular diseases . The growing knowledge about genetic influence on cardiovascular diseases ( CVD ) combined with the recently generated amounts of genomic data hold promise to the identification of new markers for atherosclerotic CVD . Cardiovascular pharmacogenomics and pharmacogenetics have now the potential for leading to identification of genetic contributors and therefore to the development of predictive genetic tests that could optimize drugs efficacy and minimize toxicity . Clinical studies have shown that genetic variations within cytochromes P450 ( CYPs ) , 3-Hydroxyl-3-Methylglutaryl DB01992 Reductase ( P04035 ) and apolipoprotein E ( P02649 ) genes influence individual 's response to lipid lowering statins . Furthermore , development of antagonists or inhibitors of molecules such as peroxisome proliferator-activated receptors ( PPARs ) , lipoprotein-associated phospholipase A(2) ( Lp-PLA(2) ) , angiotensin-converting enzyme ( P12821 ) , angiotensin receptors and tumor necrosis factor ( P01375 ) -alpha could be another alternative to prevent atherosclerosis . In addition , novel molecules under the name of biologics including family of peptides such as atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) , urocortin , apelin and antimicrobial peptides ( AMPs ) could be considered as new targets for the prevention and treatment of CVD . In this article , we will focus mainly on recent genomic advances in the development of new markers and therapeutic agents for CVD . We present an array of molecules that could have pharmacological benefit for the treatment of heart disease . We also discuss in details new strategies including biologics , which are actually the focus of companies for clinical development of therapeutic drugs . All these efforts provide optimism and attractive promise to cure CVD . P62158 interacts with angiotensin-converting enzyme-2 ( Q9BYF1 ) and inhibits shedding of its ectodomain . P12821 -2 ( Q9BYF1 ) is a regulatory protein of the renin-angiotensin system ( DB01367 ) and a receptor for the causative agent of severe-acute respiratory syndrome ( P49591 ) , the P49591 -coronavirus . We have previously shown that Q9BYF1 can be shed from the cell surface in response to phorbol esters by a process involving P01375 converting enzyme ( P78536 ; P78536 ) . In this study , we demonstrate that inhibitors of calmodulin also stimulate shedding of the Q9BYF1 ectodomain , a process at least partially mediated by a metalloproteinase . We also show that calmodulin associates with Q9BYF1 and that this interaction is decreased by calmodulin inhibitors . Granulocyte macrophage-colony stimulating factor increases the expression of histamine and histamine receptors in monocytes/macrophages in relation to arteriosclerosis . OBJECTIVE : To study the effect of granulocyte macrophage-colony-stimulating factor ( GM- P04141 ) on histamine metabolism in arteriosclerosis , the expression of histidine decarboxylase ( HDC ; histamine-producing enzyme ) , histamine receptors 1 and 2 ( P35367 and P25021 ) , and GM- P04141 was investigated in human and mouse arteriosclerotic carotid arteries . Furthermore , the molecular mechanisms of GM- P04141 -induced HDC and P35367 expression in monocytic U937 cells were investigated . METHODS AND RESULTS : Immunohistochemistry showed that atherosclerotic human coronary and mouse ligated carotid arteries contained HDC-expressing macrophages . Gene expression of HDC , P35367 , P25021 , and GM- P04141 was also detected in the lesions . In U937 cells , GM- P04141 enhanced histamine secretion and gene expression of HDC and P35367 . A promoter assay showed that GM- P04141 enhanced gene transcription of HDC and P35367 but not P25021 . CONCLUSIONS : The present results indicate that HDC and HHR are expressed in arteriosclerotic lesion , and that GM- P04141 induces HDC and P35367 expression in monocytes . Locally produced histamine might participate in atherogenesis by affecting the expression of atherosclerosis-related genes in monocytes and smooth muscle cells . The presence of histamine-producing macrophages and gene expression of histamine receptors and GM- P04141 was demonstrated in arteriosclerotic lesions . In monocytic U937 cells , GM- P04141 upregulated the expression of histamine and P35367 . Coordinated expression of histamine and its receptors by GM- P04141 would participate in atherogenesis by affecting monocytic and SMC gene expression . P00797 -angiotensin system modulation : the weight of evidence . Modulation of the renin-angiotensin system is considered to be the most complete way to manage high-risk patients including those with hypertension . P12821 ( P12821 ) inhibitors are effective at reducing the morbidity and mortality of patients with overt clinical heart failure , asymptomatic left ventricular dysfunction , and uncomplicated myocardial infarction . Furthermore , recent trials like the Heart Outcomes Prevention Evaluations ( HOPE ) study and the EUropean trial on Reduction Of cardiac events with DB00790 in stable coronary Artery disease ( EUROPA ) support extending the use of P12821 inhibitors to the routine/first-line treatment of patients with an increased global cardiovascular risk . Although some investigators have seen the development of angiotensin II receptor blockers ( ARBs ) as a more effective and tolerable way of reproducing the benefits of P12821 inhibition , there remain important concerns regarding the distinct pharmacologic profiles and modes of action of these two classes of drugs . Careful evaluation of data from recent large-scale studies revealed that , unlike P12821 inhibitors , ARBs are either neutral or may actually increase rates of myocardial infarction despite similar levels of blood pressure reduction . The fact that this effect is most apparent when ARBs are compared with placebo in the absence of concomitant P12821 inhibitors suggests that differential effects on the angiotensin II type 2 ( AT(2) ) receptors may be important . Other important pharmacologic differences are also known to be present and may be of direct relevance . The weight of available evidence therefore supports the use of appropriate P12821 inhibitor regimens , although not ARBs , in the treatment of global cardiovascular risk . Ds-echinoside A , a new triterpene glycoside derived from sea cucumber , exhibits antimetastatic activity via the inhibition of NF-κB-dependent P14780 and P15692 expressions . Ds-echinoside A ( DSEA ) , a non-sulfated triterpene glycoside , was isolated from the sea cucumber Pearsonothuria graeffei . In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion , migration , invasion , and angiogenesis . In this study , we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2 , with a half-maximal inhibitory concentration ( IC₅₀ ) of 2.65 μmol/L , and suppressed Hep G2 cell adhesion , migration , and invasion in a dose-dependent manner . DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane ( P62158 ) assay in vivo . Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 ( P14780 ) , which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis . DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 ( P01033 ) , an important regulator of P14780 activation . From the results of Western blotting , the expressions of nuclear factor-kappa B ( NF-κB ) and vascular endothelial growth factor ( P15692 ) were found to be remarkably reduced by DSEA . These findings suggest that DSEA exhibits a significant anti-metastatic activity through the specific inhibition of NF-κB-dependent P14780 and P15692 expressions . High prevalence of circulating P01730 + P10747 - T-cells in patients with small abdominal aortic aneurysms . OBJECTIVE : To assess the possible role of proinflammatory P10747 - T cells in abdominal aortic aneurysms ( AAAs ) . Animal studies and human tissue studies suggest a role for interferon ( IFN ) -gamma-producing T cells in the development and progression of AAAs . METHODS AND RESULTS : Fluorescence-activated cells sorter analysis of peripheral blood samples and measurement of AAA size using sonography were performed in 101 AAA patients and 38 healthy controls . Peripheral percentages of P10747 - T cells of the CD3+ P01730 + and the CD3+CD8+ were enriched in AAA patients with 7.8+/-8.8 % and 41.9+/-15.7 % compared with healthy controls with 2.2+/-6.1 % and 24.9+/-15.5 % , respectively ( P=0.002 and P < 0.001 , respectively ) . Both P01730 + P10747 - and CD8+ P10747 - T cells produced large amounts of IFN-[gamma] and perforin . Patients with small AAAs ( < 4 cm ) showed higher peripheral levels of P01730 + P10747 - T cells than those with larger AAAs ( P=0.025 ) . Immunohistological examinations revealed Q04695 +/-17.2 % P01730 + P10747 - and 44.0+/-13.8 % CD8+ P10747 - in AAA tissue specimens with inflammatory infiltratestes . CONCLUSIONS : P01579 - and perforin-producing P10747 - T cells are present in the periphery and the vessel wall of a majority of AAAs . This observation in humans favors the concept of a T cell-mediated pathophysiology of AAAs , especially during the early development of AAAs . P00797 -angiotensin system is involved in the mechanism of increased serum asymmetric dimethylarginine in essential hypertension . Endothelium-dependent/nitric oxide ( NO ) -mediated vasodilation is impaired in hypertensive individuals . DB01686 ( DB01686 ) , an endogenous inhibitor of NO synthase , is synthesized by many types of cells including vascular endothelial cells . The serum level of DB01686 is elevated in patients with essential hypertension , but the mechanism for this increase is unknown . Therefore , the present study examined whether the renin-angiotensin system ( DB01367 ) is involved . Patients with essential hypertension [ systolic blood pressure ( BP ) > 160 mmHg and/or diastolic BP > 95 mmHg ] were randomized to an angiotensin-converting enzyme ( P12821 ) inhibitor treatment group ( perindopril , 4mg/day for 4 weeks , n = 7 ) , an angiotensin II type 1 ( AT1 ) receptor antagonist treatment group ( losartan , 50 mg/day for 4 weeks , n = 7 ) or a beta-blocker treatment group ( bisoprolol , 5 mg/day for 4 weeks , n = 7 ) . Before and after the treatment , BP , serum concentration of DB01686 and plasma concentration of P04275 ( P04275 , a biological marker of endothelial injury ) were measured . DB00790 , losartan and bisoprolol decreased BP to a similar extent , and either perindopril or losartan , but not bisoprolol , significantly decreased serum DB01686 and plasma P04275 . These findings suggest that the DB01367 may contribute to the mechanism of increased serum DB01686 as well as to the endothelial injury observed in hypertensive patients . The vasculoprotective actions of P12821 inhibitors or AT1 receptor antagonists may be explained at least in part by amelioration of the endothelial injury through a decrease in the serum DB01686 concentration . Inhibition of central angiotensin converting enzyme ameliorates scopolamine induced memory impairment in mice : role of cholinergic neurotransmission , cerebral blood flow and brain energy metabolism . Evidences indicate that inhibition of central P00797 angiotensin system ( DB01367 ) ameliorates memory impairment in animals and humans . Earlier we have reported involvement of central angiotensin converting enzyme ( P12821 ) in streptozotocin induced neurodegeneration and memory impairment . The present study investigated the role of central P12821 in cholinergic neurotransmission , brain energy metabolism and cerebral blood flow ( Q03701 ) in model of memory impairment induced by injection of scopolamine in mice . DB00790 ( 0.05 and 0.1 mg/kg , PO ) was given orally for one week before administration of scopolamine ( 3mg/kg , IP ) . Then , memory function was evaluated by Morris water maze and passive avoidance tests . Q03701 was measured by laser Doppler flowmetry . Biochemical and molecular parameters were estimated after the completion of behavioral studies . DB00747 caused impairment in memory which was associated with reduced Q03701 , acetylcholine ( ACh ) level and elevated acetylcholinesterase ( P22303 ) activity and malondialdehyde ( MDA ) level . DB00790 ameliorated scopolamine induced amnesia in both the behavioral paradigms . Further , perindopril prevented elevation of P22303 and MDA level in mice brain . There was a significant increase in Q03701 and ACh level in perindopril treated mice . However , scopolamine had no significant effect on DB00171 level and mRNA expression of angiotensin receptors and P12821 in cortex and hippocampus . But , perindopril significantly decreased P12821 activity in brain without affecting its mRNA expression . The study clearly showed the interaction between P12821 and cholinergic neurotransmission and beneficial effect of perindopril can be attributed to improvement in central cholinergic neurotransmission and Q03701 . Different responsiveness of endothelial cells to vascular endothelial growth factor and basic fibroblast growth factor added to culture media under gravity and simulated microgravity . When incubated under simulated microgravity ( s-microg ) , endothelial cells ( EC ) form tubular structures that resemble vascular intimas . This delayed formation of 3D EC structures begins between the 5th and 7th day of culturing EC under conditions of s-microg , when double-row cell assemblies become visible . With the aim of learning about this initial phase of tubular structure formation , we found that NFkappaBp65 protein content was similar in all cell populations , but gene and protein expression of phosphokinase A catalytic subunit , phosphokinase Calpha , and extracellular signal-regulated kinases 1 and 2 was altered in cells cultured under s-microg . Apoptosis remained below 30 % in all EC cultures . In contrast to controls , the 7-day-old s-microg cultures contained 3D aggregates with proliferating cells , enhanced numbers of necrotic cells , and osteopontin-negative EC as well as supernatants with reduced quantities of vascular endothelial growth factor ( P15692 ) , basic fibroblast growth factor ( P09038 ) , soluble P25942 , P29965 , intercellular adhesion molecule-1 , tumor necrosis factor receptor 2 , Q14116 , complement P01024 , and P04275 . P15692 and/or P09038 ( 10 ng/mL ) application influenced the accumulation of proteins in supernatants more profoundly under 1 g than under s-microg . These findings provide evidence that phosphokinase Calpha plays a key role in tube formation . Improving the interaction of P15692 and/or P09038 with EC under s-microg could enhance the engineering of vascular intimas . The influence of costimulation and regulatory P01730 + T cells on intestinal IgA immune responses . It is thought that IgA B-cell differentiation is highly dependent on activated P01730 + T cells . In particular , cell-cell interactions in the Peyer 's patches involving P25942 and/or P33681 / P42081 have been implicated in germinal-center formation and IgA B-cell development . Also soluble factors , such as P05112 , P05113 , P05231 , and TGF beta may be critical for IgA B-cell differentiation in vivo . Here we report on some paradoxical findings with regard to IgA B-cell differentiation and specific mucosal immune responses that we have recently made using gene knockout mice . More specifically , we have investigated to what extent absence of P01730 + T cells , relevant cytokines , or T-cell-B-cell interactions would influence IgA B-cell differentiation in vivo . Using P01730 - or P05112 -gene knockout mice or mice made transgenic for DB01281 , we found that , although specific responses were impaired , total IgA production and IgA B-cell differentiation appeared to proceed normally . However , a poor correlation was found between , on the one hand , GC formation and IgA differentiation and , on the other hand , the ability to respond to T-cell-dependent soluble protein antigens in these mice . Thus , despite the various deficiencies in P01730 + T-cell functions seemingly intact IgA B-cell development was observed . Enhanced goblet cell hyperplasia in HDC knockout mice with allergic airway inflammation . BACKGROUND : DB11320 is known to have immunoregulatory roles in allergic reactions through histamine receptor 1 ( P35367 ) , P25021 , Q9Y5N1 and Q9H3N8 . However , its role in goblet cell hyperplasia in the airways of asthma patients is yet to be clarified . OBJECTIVE : This study was designed to examine the role of histamine in goblet cell hyperplasia using histamine-deficient mice ( Hdc-/- mice ) with allergic airway inflammation . METHODS : Wild-type and Hdc-/- C57BL/6 mice were sensitized with ovalbumin ( OVA ) . After a 2-week exposure to OVA , goblet cell hyperplasia was evaluated . Cell differentials and cytokines in BALF were analyzed . The mRNA levels of P98088 and Gob-5 gene were determined quantitatively . RESULTS : The number of eosinophils in BALF increased in both the sensitized wild-type mice and Hdc-/- mice with OVA inhalation . In addition , the numbers of alveolar macrophages and lymphocytes in BALF increased significantly in the sensitized Hdc-/- mice with OVA inhalation compared to the wild-type mice under the same conditions . The concentrations of P05112 ( P05112 ) , P05113 , P35225 , Interferon-gamma ( P01579 ) , tumor necrosis factor-alpha ( P01375 ) and P60568 in the BALF all increased significantly in both groups compared to those exposed to saline . In particular , the concentration of P01375 in the Hdc-/- mice exposed to OVA was significantly higher than that in the wild-type mice under the same conditions . The mRNA levels of Gob-5 and P98088 , and the ratio of the goblet cells in the airway epithelium significantly increased in Hdc-/- mice exposed to OVA compared to wild-type mice . CONCLUSIONS : These results suggested that histamine may play a regulatory role in goblet cell hyperplasia in allergic airway inflammation . Aflatoxin B1 induces Src phosphorylation and stimulates lung cancer cell migration . AflatoxinB1 ( AFB1 ) is well known as a potent carcinogen . Epidemiological studies have shown an association between AFB1 exposure and lung cancer in humans . AFB1 can induce the mutations of genes such as tumor suppressor p53 through its metabolite AFB1-8,9-exo-epoxide , which acts as a mutagen to react with DNA . In addition , recent study demonstrates AFB1 positively regulates type I insulin-like growth factor receptor ( IGF-IR ) signaling in hepatoma cells . The current study aims to determine the effects of AFB1 on Src kinase and insulin receptor substrate ( P41252 ) in lung cancer cells and the effects of AFB1 on lung cancer cell migration . To this end , the effects of AFB1 on P41252 expression , Src , Akt , and P29323 phosphorylation were measured by Western blot analysis . The migration of lung cancer cells was detected by wound-healing assay . AFB1 downregulates P35568 but paradoxically upregulates Q9Y4H2 through positive regulation of the stability of Q9Y4H2 and the proteasomal degradation of P35568 in lung cancer cell lines A549 and P08709 -1 . In addition , AFB1 induces Src , Akt , and P27361 /2 phosphorylation . Treatment of lung cancer cells with Src inhibitor saracatinib abrogates AFB1-induced Q9Y4H2 accumulation . Moreover , AFB1 stimulates lung cancer cell migration , which can be inhibited by saracatinib . We conclude that AFB1 may upregulate Q9Y4H2 and stimulate lung cancer cell migration through Src . The TGF-β/Smad pathway induces breast cancer cell invasion through the up-regulation of matrix metalloproteinase 2 and 9 in a spheroid invasion model system . Transforming growth factor-β ( TGF-β ) has opposing roles in breast cancer progression by acting as a tumor suppressor in the initial phase , but stimulating invasion and metastasis at later stages . In contrast to the mechanisms by which TGF-β induces growth arrest , the pathways that mediate tumor invasion are not well understood . Here , we describe a TGF-β-dependent invasion assay system consisting of spheroids of MCF10A1 normal breast epithelial cells ( M1 ) and DB01367 -transformed (pre-)malignant derivatives ( M2 and M4 ) embedded in collagen gels . Both basal and TGF-β-induced invasion of these cell lines was found to correlate with their tumorigenic potential ; M4 showing the most aggressive behavior and M1 showing the least . Basal invasion was strongly inhibited by the TGF-β receptor kinase inhibitor SB-431542 , indicating the involvement of autocrine TGF-β or TGF-β-like activity . TGF-β-induced invasion in premalignant M2 and highly malignant M4 cells was also inhibited upon specific knockdown of P84022 or Q13485 . Interestingly , both a broad spectrum matrix metalloproteinase ( MMP ) inhibitor and a selective P08253 and P14780 inhibitor mitigated TGF-β-induced invasion of M4 cells , while leaving basal invasion intact . In line with this , TGF-β was found to strongly induce P08253 and P14780 expression in a P84022 - and Q13485 -dependent manner . This collagen-embedded spheroid system therefore offers a valuable screening model for TGF-β/Smad- and P08253 - and P14780 -dependent breast cancer invasion . REV-ERBα inhibits the P35354 expression in bovine uterus endometrium stromal and epithelial cells exposed to ovarian steroids . The nuclear receptor REV-ERBα ( encoded by P20393 ) has a critical role in metabolism and physiology as well as circadian rhythm . Here , we investigated the possible contribution of clock genes including P20393 to the secretion of prostaglandin F2α ( PGF2α ) from bovine uterine stromal ( USCs ) and epithelial cells ( UECs ) by modulating the expression of P35354 . The circadian oscillation of clock genes in the cells was weak compared with that reported in rodents , but the expression of O00327 , O15534 , and P20393 was changed temporally by treatment with ovarian steroids . Significant expression of clock genes including P20393 was detected in USCs exposed to progesterone . P20393 was also significantly expressed in UECs exposed to estradiol . The expression of P35354 was suppressed in USCs exposed to progesterone , while the expression was initially suppressed in UECs exposed to estradiol and then increased after long-term exposure to estradiol . O00327 knockdown with specific siRNA caused a significant decrease in the transcript levels of P20393 and P35354 in USCs , but not in UECs . The production of PGF2α also decreased in USCs after O00327 knockdown , while its level did not significantly change in UECs . The transcript level of P35354 was increased by treatment with the antagonist of REV-ERBα in both cell types , but the agonist was ineffective . In these two cell types treated with the agonist or antagonist , the PGF2α production coincided well with the P35354 expression . Collectively , these results indicate that REV-ERBα plays an inhibitory role in the expression of P35354 in both bovine USCs and UECs treated with ovarian steroids . Genome-wide association study identifies genetic determinants of warfarin responsiveness for Japanese . DB00682 is a commonly used anticoagulant , whose dose needs to be determined for each individual patient owing to large inter-individual variability in its therapeutic dose . Although several clinical and genetic variables influencing warfarin dose have been identified , uncovering additional factors are critically important for safer use of warfarin . Through a genome-wide association study , we identified single-nucleotide polymorphism ( SNP ) rs2108622 [ cytochrome P450 , family 4 , subfamily F , polypeptide 2 ( P78329 ) ] as a genetic determinant of warfarin responsiveness for Japanese . Stratifying subjects who have been pre-classified according to the genotypes of SNP rs10509680 [ cytochrome P450 , family 2 , subfamily C , polypeptide 9 ( P11712 ) ] and SNP rs9923231 [ vitamin K epoxide reductase complex subunit 1 ( Q9BQB6 ) ] , based on their genotypes of rs2108622 allowed identification of subjects who require higher dose of warfarin . Incorporating genotypes of rs2108622 into a warfarin dosing algorithm that considers age , body surface area , status of amiodarone co-administration and genotypes of SNPs in the P11712 and Q9BQB6 genes improved the model 's predictability to 43.4 % . In this study , the association of P78329 with warfarin dose of the Japanese has been established for the first time . Besides , a warfarin dosing algorithm that incorporates genotypes of rs2108622 and amiodarone co-administration status was suggested for the Japanese . Our study also implied that common SNPs other than those in the P11712 , Q9BQB6 and P78329 genes that show strong effect on the therapeutic warfarin dose might not exist . A case of primary combined neuroendocrine carcinoma with squamous cell carcinoma in the upper gingiva . Neuroendocrine carcinoma is a rare neoplasm that occurs widely in various organs and tissues . The biological behavior of this tumor in the oral region remains poorly understood . We encountered an extremely rare case of combined neuroendocrine carcinoma with squamous cell carcinoma , occurring at the buccal gingiva in a 62-year-old woman . Left partial maxillectomy was performed . Histological examinations revealed solid nests with extensive necrosis and nuclear palisading at the periphery . The tumor also showed areas of stratified neoplastic squamous differentiation . Immunohistochemically , tumor nests stained positive for synaptophysin , chromogranin , N- P62158 ( CD56 ) , and neuron-specific enolase . Strong positivity was seen for P02533 and Q04695 in the squamous component and for P08729 in the neuroendocrine component . Both components showed P08727 staining . Cells with squamous differentiation and P02533 staining occasionally expressed p63 . The patient showed no evidence of disease as of 23 months postoperatively . Given the aggressive characteristics of neuroendocrine carcinoma , strict follow-up has been performed . Deficiency of P23219 causes natriuresis and enhanced sensitivity to P12821 inhibition . BACKGROUND : Prostanoid products of the cyclo-oxygenase ( P36551 ) pathway of arachidonic acid metabolism modulate blood pressure ( BP ) and sodium homeostasis . Conventional non-steroidal anti-inflammatory drugs ( NSAIDs ) , which inhibit both P36551 isoforms ( P23219 and -2 ) , cause sodium retention , exacerbate hypertension , and interfere with the efficacy of certain anti-hypertensive agents such as angiotensin-converting enzyme ( P12821 ) inhibitors . While a new class of NSAIDs that specifically inhibit P35354 is now widely used , the relative contribution of the individual P36551 isoforms to these untoward effects is not clear . METHODS : To address this question , we studied mice with targeted disruption of the P23219 ( Ptgs1 ) gene . Blood pressure , renin mRNA expression , and aldosterone were measured while dietary sodium was varied . To study interactions with the renin-angiotensin system , P12821 inhibitors were administered and mice with combined deficiency of P23219 and the angiotensin II subtype 1A ( AT1A ) receptor were generated . RESULTS : On a regular diet , BP in P23219 -/- mice was near normal . However , during low salt feeding , BP values were reduced in P23219 -/- compared to +/+ animals , and this reduction in BP was associated with abnormal natriuresis despite appropriate stimulation of renin and aldosterone . Compared to P23219 +/+ mice , the actions of P12821 inhibition were markedly accentuated in P23219 -/- mice . Sodium sensitivity and BP lowering also were enhanced in mice with combined deficiency of P23219 and AT1A receptor . CONCLUSIONS : The absence of P23219 is associated with sodium loss and enhanced sensitivity to P12821 inhibition , suggesting that P23219 inhibition does not cause hypertension and abnormal sodium handling associated with NSAID use . Differential effects of P84022 targeting in a murine model of chronic kidney disease . Transforming growth factor ( TGF ) -β1 has a pivotal role in the pathogenesis of progressive kidney diseases that are characterized by fibrosis . The main intracellular signaling pathway of TGF-β1 is the Smad system , where Q15796 and P84022 play a central role in transcriptional regulation of target genes involved in extracellular matrix ( Q13201 ) metabolism . This study analyzes the hypothesis that blockade of P84022 attenuates the development of TGF-β1-driven renal fibrosis . This was examined in vivo in a transgenic model of TGF-β1-induced chronic kidney disease with P84022 or without P84022 expression and in vitro in mesangial cells and glomerular endothelial cells with Q15796 /3 inhibitors or P84022 -knockdown . Electron microscopy was used for evaluation of morphological changes , real-time polymerase chain reaction for detection of RNA expression , and immunohistochemistry for localization of Q13201 components . Matrix metalloproteinase ( MMP ) level was assessed by gelatin zymography electrophoresis and located by in situ zymography . The results show TGF-β1-induced mesangial matrix expansion , tubulointerstitial fibrosis , and tubular basement membrane thickening that are attenuated by P84022 deletion , whereas TGF-β1-induced glomerular basement membrane thickening is not shown . The amount and distribution profile of P08253 may suggest a role of the enzyme herein . We conclude that P84022 targeting is not exclusively beneficial as P84022 has diverse transcriptional regulatory effects in different cell types in the kidney . Increased myocardial collagen content in transgenic rats overexpressing cardiac angiotensin-converting enzyme is related to enhanced breakdown of N-acetyl- DB00133 - DB00128 -Lys-Pro and increased phosphorylation of Q15796 /3 . BACKGROUND : Although increased activity of angiotensin-converting enzyme ( P12821 ) has been associated with increased cardiac collagen , no studies to date have established a direct cause-and-effect relation between the two . METHODS AND RESULTS : We used transgenic rats that overexpress human P12821 selectively in the myocardium . Two independent heterozygous transgenic rat lines were studied , one expressing 2 to 3 copies ( L1172 ) and the other expressing 5 to 10 copies ( L1173 ) of the P12821 transgene . These rats were normotensive but developed a proportionate increase in myocardial collagen depending on the P12821 gene dose ( up to 2.5-fold , P < 0.01 ) , but cardiac angiotensin II levels remained normal , whereas collagen content reversed to control levels on P12821 inhibition . To explain these changes , we investigated N-acetyl- DB00133 - DB00128 -Lys-Pro ( AcSDKP ) , an alternative substrate that is catabolized exclusively by P12821 . Increased cardiac expression of P12821 was paralleled by a reciprocal decrease in cardiac AcSDKP and a proportionate increase in phosphorylated Q15796 and P84022 , all of which normalized after both P12821 inhibition and AcSDKP infusion . Furthermore , a functional link of this signaling cascade was demonstrated , because AcSDKP inhibited P84022 phosphorylation in a dose-dependent manner in cultured cardiac fibroblasts and in vivo . CONCLUSIONS : Our findings suggest that increased cardiac P12821 activity can increase cardiac collagen content by degradation of AcSDKP , an inhibitor of the phosphorylation of transforming growth factor-beta signaling molecules Q15796 and P84022 . This implies that the antifibrotic effects of P12821 inhibitors are mediated in part by increasing cardiac AcSDKP , with subsequent inhibition of Smad 2/3 phosphorylation . Female , but not male , mice show delayed cutaneous wound healing following aspirin administration . Cyclo-oxygenase ( P36551 ) is an enzyme that participates in the wound healing process . DB00945 , a non-steroidal anti-inflammatory drug , simultaneously inhibits the aromatase activity of P23219 and P35354 isoforms , which is needed for prostaglandin synthesis . The aim of the present study was to determine whether aspirin , and thus P36551 inhibition , distinctly affects cutaneous wound healing in female and male mice . Female and male BALB/c mice were treated with aspirin ( 25 mg/kg per day ) for 16 days until they were killed . The control group received vehicle ( saline ) only . A full-thickness excisional lesion was made on the back , 2 days after aspirin administration started , and macroscopic , histological and biochemical parameters were evaluated . Sections were stained and immunostained for microscopic analysis . P05164 ( P05164 ) activity , hydroxyproline quantity and the protein expression of P04275 ( P04275 ) and vascular endothelial growth factor ( P15692 ) were also determined . Female control and aspirin-treated groups exhibited delayed wound closure and re-epithelization compared with the male control and aspirin-treated groups , respectively . The female control group exhibited reduced P05164 activity and a decreased number of macrophage inhibitory factor-positive cells compared with the male control group . In the female aspirin-treated group , P05164 activity and the number of F4/80-positive macrophages was higher than in the control group . Collagen was reduced only in the female aspirin-treated group . The expression of P04275 and P15692 protein was increased in the female aspirin-treated group . In conclusion , aspirin administration impaired the wound healing process in BALB/c female , but not male , mice . Potent antitumor efficacy of interleukin-18 delivered by conditionally replicative adenovirus vector in renal cell carcinoma-bearing nude mice via inhibition of angiogenesis . It has been demonstrated that interleukin 18 ( Q14116 ) exerts antitumor activity . In this study , we investigated whether oncolytic adenovirus-mediated gene transfer of Q14116 could induce strong antitumor activity . A tumor-selective replicating adenovirus expressing Q14116 ( ZD55- Q14116 ) was constructed by insertion of an Q14116 expression cassette into the ZD55 vector , which is based on deletion of the adenoviral E1B 55-kDa gene . ZD55- Q14116 could express substantially more Q14116 than Ad- Q14116 because of replication of the vector . It has been shown that ZD55- Q14116 exerted a strong cytopathic effect and significant apoptosis in renal cell carcinoma . ZD55- Q14116 significantly decreased P15692 and P28906 expression in the tumor cells . Treatment of established tumors with ZD55- Q14116 showed much stronger antitumor activity than that induced by ZD55-EGFP or Ad- Q14116 . These data indicated that oncolytic adenovirus expressing Q14116 could exert potential antitumor activity via inhibition of angiogenesis and offer a novel approach to cancer therapy . Different effects of perindopril and enalapril on monocyte cytokine release in coronary artery disease patients with normal blood pressure . BACKGROUND : Favorable effects of angiotensin-converting enzyme ( P12821 ) inhibitor treatment on the incidence of cardiovascular and cerebrovascular mortality and morbidity are not limited to patients with elevated blood pressure . As suggested by our previous results , the physicochemical and pharmacokinetic differences between drugs may markedly contribute to the strength of pleiotropic effects of P12821 inhibitors . METHODS : The present study was aimed at comparing the effects of serum- and tissue-type P12821 inhibitors on monocyte release of proinflammatory cytokines in normotensive patients with stable coronary artery disease . The participants were randomized to 90-day treatment with enalapril ( 20 mg daily , n = 29 ) , perindopril ( 4 mg daily , n = 27 ) or placebo ( n = 28 ) . Plasma levels of lipids , glucose , insulin and high sensitivity P02741 ( hsCRP ) , as well as monocyte release of proinflammatory cytokines were determined before and after 30 days of therapy , and at the end of the treatment . RESULTS : Lipopolysaccharide-stimulated monocytes from normotensive patients with stable coronary artery disease released significantly more P01375 -α , interleukin-1β and monocyte chemoattractant protein-1 in comparison with monocytes from 23 matched control subjects . Their baseline hsCRP levels were also higher . DB00790 reversed the disease-induced changes in cytokine release and reduced plasma hsCRP , while the effect of enalapril was much more limited . The effect on both drugs on cytokine release was stronger in insulin-resistant than insulin-sensitive subjects . CONCLUSIONS : Our results indicate that perindopril is superior to enalapril in producing monocyte-suppressing and systemic anti-inflammatory effects in normotensive patients with coronary artery disease . This action may contribute to the clinical effectiveness of tissue P12821 inhibitors in the therapy of atherosclerosis-related disorders , particularly in insulin-resistant subjects . Pressure overload induces Q14116 and IL-18R expression , but markedly suppresses O95998 expression in a rabbit model . Q14116 potentiates P01375 -α-induced cardiomyocyte death . Recurrent or sustained inflammation plays a causal role in the development and progression of left ventricular hypertrophy ( LVH ) and its transition to failure . Interleukin ( IL ) -18 is a potent pro-hypertrophic inflammatory cytokine . We report that induction of pressure overload in the rabbit , by constriction of the descending thoracic aorta induces compensatory hypertrophy at 4weeks ( mass/volume ratio : 1.7±0.11 ) and ventricular dilatation indicative of heart failure at 6weeks ( mass/volume ratio : 0.7±0.04 ) . In concordance with this , fractional shortening was preserved at 4weeks , but markedly attenuated at 6weeks . We cloned rabbit Q14116 , IL-18Rα , IL-18Rβ , and Q14116 binding protein ( O95998 ) cDNA , and show that pressure overload , while enhancing Q14116 and IL-18R expression in hypertrophied and failing hearts , markedly attenuated the level of expression of the endogenous Q14116 antagonist O95998 . Cyclical mechanical stretch ( 10 % cyclic equibiaxial stretch , 1Hz ) induced hypertrophy of primary rabbit cardiomyocytes in vitro and enhanced P01160 , Q14116 , and IL-18Rα expression . Further , treatment with rhIL-18 induced its own expression and that of IL-18Rα via AP-1 activation , and induced cardiomyocyte hypertrophy in part via PI3K/Akt/ P43694 signaling . In contrast , Q14116 potentiated P01375 -α-induced cardiomyocyte death , and by itself induced cardiac endothelial cell death . These results demonstrate that pressure overload is associated with enhanced Q14116 and its receptor expression in hypertrophied and failingrabbit hearts . Since O95998 expression is markedly inhibited , our results indicate a positive amplification in Q14116 proinflammatory signaling during pressure overload , and suggest Q14116 as a potential therapeutic target in pathological hypertrophy and cardiac failure . Progressive encephalomyelitis with rigidity presenting as a stiff-person syndrome . Diagnosis criteria of stiff-person syndrome ( P49903 ) include progressive , fluctuating muscular rigidity and spasms with normal neurological examination . The presence of unusual features such as prominent limb rigidity with segmental signs and contracture , evidence of brainstem dysfunction , profound autonomic disturbances , P04141 pleiocytosis or Q9BWK5 abnormalities in patients with P49903 presentation allows to classify these patients as progressive encephalomyelitis with rigidity ( O15534 ) . We report a 50 year-old woman suffering from severe painful spasms of abdominal wall and limb muscles . Neurological examination showed pyramidal signs . EMG disclosed continuous muscle activity with superimposed discharges . Treatment with high doses of diazepam and baclofen led to moderate improvement of generalised stiffness . However , the right arm became more rigid with oedema and vasomotor changes . Subsequently , bilateral nystagmus and internuclear opthalmplegia appeared . There was mild P04141 pleiocytosis . Associated auto-immune thyroiditis was found with positive anti-microsome antibodies and decreased thyroid hormones . Search for profound neoplasm was negative . The patient had three subacute bouts then she improved with methylprednisolone . The initial clinical presentation mimicking a P49903 with subsequent diffuse involvement of the central nervous system and a striking localisation of a severe rigidity to one arm allowed to suspect the diagnosis of O15534 . The relationship between P49903 and O15534 remains unclear because of the rarity of these disorders . The observation reported in this paper gives evidence that both the disorders are probably two clinical presentations of the same pathogenic process . Cardiac protective effect of Astragalus on viral myocarditis mice : comparison with DB00790 . In clinical practice , Astragali Radix ( Astragalus ) , the root of Astragalus membranaceus Bunge , has been widely applied to treat patients with viral diseases , including viral myocarditis in China . The present study was designed to evaluate the protective effects of Astragalus on the function of sarcoplasmic reticulum calcium ATPase ( P16615 ) activity and endothelin system at acute and chronic periods of myocarditis mice induced by CVB(3) infection . Astragalus feeding ( 2.2 mg/kg/day ) could significantly increase the survival rate , alleviate pathological alterations and serum cardiac troponin I ( cTnI ) , as well as restore impaired SERCA activity at the acute stage . Low affinity and capacity of ETR were reversed with Astragalus after the first CVB(3) inoculation up to 7 days and after the second virus inoculation up to 150 days . In the meantime , the contents of cardiac ET-1 and P01160 were reduced . Comparison the myocarditis mice treated with DB00790 ( 0.44 mg/kg/day ) , an P12821 inhibitor , shows that Astragalus achieved a similar effect on survival rate , P16615 and ET system . These results indicated that the beneficial effects of Astragalus and DB00790 for treating viral myocarditis might be partly mediated by preserving the functions of SERCA 2 activity and ET system . Role of angiotensin II in the remodeling induced by a chronic increase in flow in rat mesenteric resistance arteries . Angiotensin II is a potent growth factor involved in arterial wall homeostasis . In resistance arteries , chronic increases in blood flow induce a rise in diameter associated with arterial wall hypertrophy . Nevertheless , the role of angiotensin II in this remodeling is unknown . We investigated the effect of blocking angiotensin II production or receptor activation on flow-induced remodeling of mesenteric resistance arteries . Arteries were ligated in vivo to generate high-flow arteries compared with normal flow ( control ) vessels located at a distance . Arteries were isolated after 1 week for in vitro analysis . Arterial diameter , media surface , endothelial NO synthase expression , superoxide production , and extracellular signal-regulated kinase 1/2 phosphorylation were higher in high-flow than in control arteries . P12821 inhibition ( perindopril ) and angiotensin II type 1 receptor blockade ( candesartan ) prevented arterial wall hypertrophy without affecting diameter enlargement . The nonselective vasodilator hydralazine had no effect on remodeling . Although perindopril and candesartan increased endothelial NO synthase expression in high-flow arteries , hypertrophy remained in rats treated with N(G)-nitro-l-arginine methyl ester and mice lacking endothelial NO synthase . DB00790 and candesartan reduced oxidative stress in high-flow arteries , but superoxide scavenging did not prevent hypertrophy . Both Tempol and the absence of endothelial NO synthase prevented the rise in diameter in high-flow vessels . P27361 /2 activation in high-flow arteries was prevented by perindopril and candesartan and not by hydralazine . P27361 /2 inhibition in vivo ( U0126 ) prevented hypertrophy in high-flow arteries . Thus , a chronic rise in blood flow in resistance arteries induces a diameter enlargement involving NO and superoxide , whereas hypertrophy was associated with extracellular signal-regulated kinase 1/2 activation by angiotensin II . Inhibition of the renin-angiotensin system improves physiological outcomes in mice with mild or severe cancer cachexia . Cancer cachexia describes the progressive skeletal muscle wasting and weakness associated with many cancers . Cachexia reduces mobility and quality of life and accounts for 20-30 % of all cancer-related deaths . Activation of the renin-angiotensin system causes skeletal muscle wasting and weakness . We tested the hypothesis that treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , perindopril , would enhance whole body and skeletal muscle function in cachectic mice bearing Colon-26 ( C-26 ) tumors . CD2F1 mice received a subcutaneous injection of phosphate buffered saline or C-26 tumor cells inducing either a mild or severe cachexia . The following day , one cohort of C-26 mice began receiving perindopril in their drinking water ( 4 mg kg(-1) day(-1) ) for 21 days . In mild and severe cachexia , perindopril increased measures of whole body function ( grip strength and rotarod ) and reduced fatigue in isolated contracting diaphragm muscle strips ( p < 0.05 ) . In severely cachectic mice , perindopril reduced tumor growth , improved locomotor activity and reduced fatigue of tibialis anterior muscles in situ ( p < 0.05 ) , which was associated with increased oxidative enzyme capacity ( succinate deyhydrogenase , p < 0.05 ) . DB00790 attenuated the increase in Q969Q1 and P05231 mRNA expression and enhanced Akt phosphorylation in severely cachectic mice but neither body nor muscle mass was increased . These findings support the therapeutic potential of P12821 inhibition for enhancing whole body function and reducing fatigue of respiratory muscles in early and late stage cancer cachexia and should be confirmed in future clinical trials . Since P12821 inhibition alone did not enhance body or muscle mass , co-treatment with an anabolic agent may be required to address these aspects of cancer cachexia . Impaired endothelial antithrombotic activity following short-term interruption of continuous subcutaneous insulin infusion in type 1 diabetic patients . Review of literature has shown an increased rate of thrombotic complications in diabetic patients with frequent episodes of hyperketonemia . However , the mechanisms by which ketosis promotes vascular disease in diabetic patients are unclear . It was the aim of this study to investigate early changes in haemostatic parameters and oxidative stress markers during the hyperketonemic status which follows the interruption of continuous subcutaneous insulin infusion ( CSII ) in type I diabetic patients . Eight CSII-treated type I diabetic patients underwent a 4-hour pump arrest . Blood glucose , insulin and 3-hydroxybutirate were measured to verify the metabolic response . A vein-occlusive ( VO ) test was performed for the determination of tPA and P05121 activities and their antigen levels before and after the CSII arrest . P08709 and VIII were evaluated by one-stage PT and PTT method , respectively . TF , P04275 , tPA and P05121 antigens were determined by ELISA , whereas tPA and P05121 activities using chromogenic methods . Plasma malondialdehyde ( MDA ) and protein carbonyl groups ( DB01053 ) levels were determined by HPLC and spectrophotometry , respectively . After the insulin deprivation phase , post-VO tPA antigen level significantly decreased ( P = 0.0391 ) , whereas TF and post-VO P05121 activity and antigen levels significantly increased ( P = 0.0156 and P = 0.0234 , respectively ) . Plasma MDA and DB01053 levels were 1.88-fold and 1.74-fold higher than baseline values , respectively . In conclusion , the impairment of the fibrinolytic potential and the increases in TF , MDA and DB01053 levels may enhance the risk of both arterial and venous thrombosis during ketosis . Thus , early detection of hyperketonemia in DM patients could contribute to the prevention of life-threatening vascular events . Effects of ellagic Acid on angiogenic factors in prostate cancer cells . BACKGROUND : Several natural antioxidants , including ellagic acid ( EA ) , have been reported to have chemotherapeutic activity in vivo and in vitro settings . Cytochrome P450 ( CYP ) activity and synthesis of both epoxyeicosatrienoic acids ( EETs ) and 20-hydroxy-5,8,11,14-eicosatetraenoic acid ( 20-HETE ) , together with vascular endothelial growth factor ( P15692 ) and heme oxygenase system ( HO ) have emerged as important modulators of tumor growth and metastasis . METHODS : The anti-angiogenic effects of EA were investigated in the human prostatic cancer cell line LnCap . P09601 , P30519 , P51589 and soluble epoxyde hydrolase ( sEH ) expressions were evaluated by western blotting . Levels of P15692 and osteoprotegerin ( O00300 ) were determined in the culture supernatant using an ELISA assay , while CYP mRNAs were determined by qRT-PCR . RESULTS : EA treatment induced a significant decrease ( p < 0.05 ) in P09601 , P30519 and P51589 expression , and in P15692 and O00300 levels . Similarly P51589 , P78329 and CYPA22 mRNAs were significantly ( p < 0.05 ) down-regulated by EA treatment . The decrease in P51589 mRNA was associated with an increase in sEH expression . CONCLUSIONS : RESULTS reported in the present study highlighted the ability of EA to modulate a new pathway , in addition to anti-proliferative and pro-differentiation properties , via a mechanism that involves a decrease in eicosanoid synthesis and a down-regulation of the HO system in prostate cancer . P12821 activity is involved in the mechanism of increased endogenous nitric oxide synthase inhibitor in patients with type 2 diabetes mellitus . The renin-angiotensin system plays an important role in the elevation of asymmetric dimethylarginine ( DB01686 ) , an endogenous inhibitor of nitric oxide synthase , in hypertensive patients , so the present study was designed to examine whether angiotensin-converting enzyme ( P12821 ) activity is also involved in the mechanism of DB01686 elevation in type 2 diabetes mellitus ( NIDDM ) . A crossover study was performed to determine if P12821 inhibition with perindopril ( 4 mg/day ) for 4 weeks decreases serum DB01686 concentration and plasma P04275 ( P04275 ) level ( a marker of endothelial injury ) in 11 patients with NIDDM . None of the patients was treated with insulin or oral hypoglycemic drugs , and none had major diabetic complications . Before the protocol began , serum DB01686 and plasma P04275 were significantly higher in the 11 NIDDM patients , when compared with 8 control subjects without diabetes . DB00790 did not affect blood pressure or glucose metabolism , but did significantly decrease serum DB01686 and plasma P04275 . These results suggest that endothelial injury associated with DB01686 elevation may be present even in patients with non-complicated NIDDM , and that increased activity of P12821 may be involved in such endothelial dysfunction . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Gene expression profiling reveals renin mRNA overexpression in human hypertensive kidneys and a role for microRNAs . The kidney has long been invoked in the etiology of essential hypertension . This could involve alterations in expression of specific genes and microRNAs ( miRNAs ) . The aim of the present study was to identify , at the transcriptome-wide level , mRNAs and miRNAs that were differentially expressed between kidneys of 15 untreated hypertensive and 7 normotensive white male subjects of white European ancestry . By microarray technology we found 14 genes and 11 miRNAs that were differentially expressed in the medulla . We then selected and confirmed by real-time quantitative PCR expression differences for P22736 , P43354 , Q92570 , O15534 , and P57059 mRNAs and for the miRNAs hsa-miR-638 and hsa-let-7c . Luciferase reporter gene experiments in human kidney ( HEK293 ) cells confirmed the predicted binding of hsa-let-7c to the 3' untranslated region of P43354 mRNA . In the renal cortex we found differential expression of 46 genes and 13 miRNAs . We then confirmed expression differences for O95831 , P02760 , P02649 , P16671 , P98172 , Q9Y375 , P30044 , REN , P51606 , Q9BZW2 , Q12846 , and P45379 mRNAs and for miRNAs hsa-miR-21 , hsa-miR-126 , hsa-miR-181a , hsa-miR-196a , hsa-miR-451 , hsa-miR-638 , and hsa-miR-663 . Functional experiments in HEK293 cells demonstrated that hsa-miR-663 can bind to the REN and P02649 3' untranslated regions and can regulate REN and P02649 mRNA levels , whereas hsa-miR-181a regulated REN and O95831 mRNA . Our data demonstrated for the first time that miRNAs can regulate renin expression . The observed downregulation of 2 miRNAs in hypertension could explain the elevation in intrarenal renin mRNA . P00797 , P16671 , and other mRNAs , as well as miRNAs and associated pathways identified in the present study , provide novel insights into hypertension etiology . TGF-β-elicited induction of tissue inhibitor of metalloproteinases ( P01033 ) -3 expression in fibroblasts involves complex interplay between P84022 , p38α , and P27361 /2 . Transforming growth factor-β ( TGF-β ) promotes extracellular matrix deposition by down-regulating the expression of matrix degrading proteinases and upregulating their inhibitors . Tissue inhibitor of metalloproteinases ( P01033 ) -3 is an Q13201 -associated specific inhibitor of matrix degrading metalloproteinases . Here , we have characterized the signaling pathways mediating TGF-β-induced expression of P35625 . Basal and TGF-β-induced P35625 mRNA expression was abolished in Q13485 -deficient mouse embryonic fibroblasts and restoring Q13485 expression rescued the response . Inhibition of Smad signaling by expression of O15105 and dominant negative P84022 completely abolished TGF-β-elicited expression of P35625 in human fibroblasts , whereas overexpression of P84022 enhanced it . Inhibition of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) activation with PD98059 and p38 mitogen-activated protein kinase activity by SB203580 resulted in suppression of TGF-β-induced P35625 expression , indicating that P27361 /2 and p38 MAPK mediate the effect of TGF-β on P35625 expression . Specific activation of p38α and P27361 /2 by constitutively active mutants of MKK3b or Q02750 , respectively , and simultaneous co-expression of P84022 resulted in induction of P35625 expression in the absence of TGF-β indicating that P84022 co-operates with p38 and P27361 /2 in the induction of P35625 expression . These results demonstrate the complex interplay between P84022 , p38α , and P27361 /2 signaling in the regulation of P35625 gene expression in fibroblasts , which may play a role in inflammation , tissue repair , and fibrosis . Differential induction of matrix metalloproteinase 1 and 2 in ectopic endometrium . According to the transplantation theory , endometriosis develops from endometrial fragments that are retrogradely menstruated into the peritoneal cavity . In order to develop into endometriotic lesions , they have to connect to the vascular system by angiogenesis , probably involving matrix metalloproteinases ( MMP ) as key enzymes in extracellular matrix remodelling . A model of endometriosis using the chorioallantoic membrane ( P62158 ) of chick embryos was established . Eutopic endometrium from healthy women was transferred to the P62158 and cultivated ectopically for up to 3 days . Before transplantation and after 24 , 48 and 72 h of culture on the P62158 , total RNA was extracted and reverse transcribed . Human P03956 ( interstitial collagenase ) and P08253 ( gelatinase A ) mRNA expression was assessed by competitive PCR . Results were normalized to the content of human glyceraldehyde 3-phosphate dehydrogenase ( P04406 ) mRNA . In eutopic endometrium , 0.29 amol P03956 mRNA and 0.42 fmol P08253 mRNA per fmol P04406 mRNA were found . Relative P03956 mRNA concentrations increased strongly after culture on the P62158 , while P08253 mRNA levels were nearly unaltered . This differential regulation suggests different roles of these enzymes in the angiogenesis of ectopic endometrial fragments and during the development of endometriosis . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . The use of microcalorimetry and HPLC for the determination of degradation kinetics and thermodynamic parameters of DB00790 Erbumine in aqueous solutions . DB00790 Erbumine ( O15534 ) is one of the newly used angiotensin-converting enzyme inhibitors ( P12821 inhibitors ) and is used for the treatment of patients with hypertension and symptomatic heart failure . It has two main degradation pathways , i.e. the degradation by hydrolysis and the degradation by cyclization . An isothermal heat conduction microcalorimetry ( MC ) and high pressure liquid chromatography ( HPLC ) were used for the characterization of aqueous solutions of O15534 and its stability properties . The rates of heat evolved during degradation of perindopril were measured by MC as a function of temperature and pH and from these data rate constant and change in enthalpy of the reactions were determined . With the HPLC method the concentration of perindopril and its degradation products were measured as a function of time in aqueous solutions of different pH that were stored at different temperatures . We demonstrated that reactions of degradation of perindopril at observed conditions follow the first order kinetics . The Arrhenius equation for each pH was determined . At pH 6.8 only one degradation pathway is present , i.e. the degradation by hydrolysis . Degradation constants for this pathway calculated from MC data are in good agreement with those obtained from HPLC . MC as a non-specific technique was shown to be useful in studies of O15534 when one reaction was present in the sample and also when more chemical and physical processes were simultaneously running . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . [ An effect of perindopril on the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 in peripheral blood in the acute period of atherothrombotic ischemic stroke and myocardial infarction ] . Twenty-nine patients with acute atherothrombotic ischemic stroke and 36 patients with acute Q-wave myocardial infarction have been studied . Each group has been stratified into 2 subgroups : patients of subgroups A received an P12821 inhibitor perindopril in the complex therapy from the 1st day of disease . Patients of subgroups B were not assigned to this drug . Along with routine tests , the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 ( P14780 ) measured with ELISA using test-systems ( Q02223 Diagnostics , USA ) and reagents ( R & D , England ) have been determined . The administration of perindopril did not cause side-effects , including arterial hypotonia after the first dosage , in patients in the acute period of atherothrombotic ischemic stroke and myocardial infarction . DB00790 may decrease the activity of P14780 in these patients and produces an anticytokine effect . Some similar mechanisms of ischemic lesions of the heart and the brain and a commonness of biochemical " response " to the same medical intervention ( the administration of an P12821 inhibitor perindopril ) in patients of both groups were found . The results support the pathogenetic validity of perindopril therapy in the secondary prevention of ischemic stroke and myocardial infarction . Experimental autoimmune encephalomyelitis in the Wistar rat : dependence of MBP-specific T cell responsiveness on P33681 costimulation . Experimental autoimmune encephalomyelitis ( EAE ) is an animal model of human multiple sclerosis that requires the activation of autoreactive T cells for the expression of pathology . EAE has been most frequently studied in the Lewis rat model as well as in several murine models of EAE including the PLJ and B10PL strains . In the present study we describe a novel model of EAE induced in the Wistar rat strain by immunization with guinea pig spinal cord antigens and pertussis toxin ( PT ) . T cell responses were induced to myelin basic protein . Autoreactive T cells could be totally blocked by the in vitro treatment with DB01281 , a protein that blocks the costimulation of autoreactive T cells . The addition of P60568 could reverse the inhibition seen in vitro with DB01281 . The effects of inhibition of P33681 costimulation were also examined by an analysis of cytokine responses and P60568 receptor on T cells . DB01281 treatment in vitro reduced the expression of P60568 receptor on T cells , enhanced T cell apoptosis and decreased the synthesis of P60568 , P01579 and P01375 . DB01281 treatment had no effect on P22301 synthesis by T cells , a cytokine implicated in the functions of regulatory T cell subsets . Overall , our studies support the rationale of P33681 blocking therapies as a potential treatment for models of multiple sclerosis . The induction of EAE in the Wistar rat provides yet another novel model in which to examine the regulation of T cell autoimmunity . Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 and an inducible form termed P35354 . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 and P35354 . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 with the P23219 inhibitor and an R0 of 21 A for P35354 with the P35354 inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 and 18 A in P35354 . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange .

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interacts_with DB08918?

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Reticulons O95197 and Q9NQC3 -B/C interact with P56817 and inhibit its ability to produce amyloid beta-protein . Beta-secretase beta-site P05067 cleaving enzyme 1 ( P56817 ) , is a membrane-bound aspartyl protease necessary for the generation of amyloid beta-protein ( Abeta ) , which accumulates in the brains of individuals with Alzheimer 's disease ( AD ) . To gain insight into the mechanisms by which P56817 activity is regulated , we used proteomic methods to search for P56817 -interacting proteins in human neuroblastoma SH-SY5Y cells , which overexpress P56817 . We identified reticulon 4-B ( Q9NQC3 -B ; Nogo-B ) as a P56817 -associated membrane protein . Co-immunoprecipitation experiments confirmed a physical association between P56817 and Q9NQC3 -B , Q9NQC3 -C ( the shortest isoform of RTN-4 ) , and their homologue reticulon 3 ( O95197 ) , both in SH-SY5Y cells and in transfected human embryonic kidney ( P29320 ) 293 cells . Overexpression of these reticulons ( RTNs ) resulted in a 30-50 % reduction in the secretion of both Abeta40 and Abeta42 from HEK293 cells expressing the AD-associated Swedish mutant amyloid precursor protein ( P05067 ) , but did not affect Abeta secretion from cells expressing the P05067 beta-C-terminal fragment ( beta-CTF ) , indicating that these RTNs can inhibit P56817 activity . Furthermore , a P56817 mutant lacking most of the N-terminal ectodomain also interacted with these RTNs , suggesting that the transmembrane region of P56817 is critical for the interaction . We also observed a similar interaction between these RTNs and the P56817 homologue Q9Y5Z0 . Because O95197 and Q9NQC3 -B/C are substantially expressed in neural tissues , our findings suggest that they play important roles in the regulation of P56817 function and Abeta production in the brain . No replication of genetic association between candidate polymorphisms and Alzheimer 's disease . Alzheimer 's disease is a genetically complex disorder , for which new putative susceptibility genes are constantly proposed in the literature . We selected 16 candidate genes involved in biological pathways closely related to the pathology , and for which a genetic association with Alzheimer 's disease was previously detected : P12821 , P56817 , P23560 , P42892 , P98160 , P14735 , IL1a , P05231 , P22301 , P10636 , P00749 , PrnP , P49768 , Q92673 , Q12800 and TGFb1 . The variants originally associated with the disease were genotyped in a French Caucasian sample including 428 cases and 475 controls and tested for association in order to replicate the initial results . Despite a careful replication study design , we failed to validate the initial findings for any of these variants , with the possible exception of P10636 , Q92673 and Q12800 for which some nominal but inconsistent evidence of association was observed . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . α-Asarone Ameliorates Memory Deficit in Lipopolysaccharide-Treated Mice via Suppression of Pro-Inflammatory Cytokines and Microglial Activation . α-Asarone exhibits a number of pharmacological actions including neuroprotective , anti-oxidative , anticonvulsive , and cognitive enhancing action . The present study investigated the effects of α-asarone on pro-inflammatory cytokines mRNA , microglial activation , and neuronal damage in the hippocampus and on learning and memory deficits in systemic lipopolysaccharide ( LPS ) -treated C57BL/6 mice . Varying doses of α-asarone was orally administered ( 7.5 , 15 , or 30 mg/kg ) once a day for 3 days before the LPS ( 3 mg/kg ) injection . α-Asarone significantly reduced P01375 -α and IL-1β mRNA at 4 and 24 hours after the LPS injection at dose of 30 mg/kg . At 24 hours after the LPS injection , the loss of P00915 neurons , the increase of TUNEL-labeled cells , and the up-regulation of P56817 expression in the hippocampus were attenuated by 30 mg/kg of α-asarone treatment . α-Asarone significantly reduced Iba1 protein expression in the hippocampal tissue at a dose of 30 mg/kg . α-Asarone did not reduce the number of Iba1-expressing microglia on immunohistochemistry but the average cell size and percentage areas of Iba1-expressing microglia in the hippocampus were significantly decreased by 30 mg/kg of α-asarone treatment . In the Morris water maze test , α-asarone significantly prolonged the swimming time spent in the target and peri-target zones . α-Asarone also significantly increased the number of target heading and memory score in the Morris water maze . The results suggest that inhibition of pro-inflammatory cytokines and microglial activation in the hippocampus by α-asarone may be one of the mechanisms for the α-asarone-mediated ameliorating effect on memory deficits . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Ceramide stabilizes beta-site amyloid precursor protein-cleaving enzyme 1 and promotes amyloid beta-peptide biogenesis . The lipid second messenger ceramide regulates several biochemical events that occur during aging . In addition , its level is highly elevated in the amyloid-burdened brains of Alzheimer 's disease patients . Here , we analyzed the impact of aberrant ceramide levels on amyloid beta-peptide ( Abeta ) generation by using a cell-permeable analog of ceramide , P13671 -ceramide , and several biochemical inhibitors of the sphingomyelin/glycosphingolipid biosynthetic pathway . We found that P13671 -ceramide increased the biogenesis of Abeta by affecting beta-but not gamma-cleavage of the amyloid precursor protein . Similarly to P13671 -ceramide , increased levels of endogenous ceramide induced by neutral sphingomyelinase treatment also promoted the biogenesis of Abeta . Conversely , fumonisin B1 , which inhibits the biosynthesis of endogenous ceramide , reduced Abeta production . Exogenous P13671 -ceramide restored both intracellular ceramide levels and Abeta generation in fumonisin B1-treated cells . These events were specific for amyloid precursor protein and were not associated with apoptotic cell death . Pulse-chase and time-course degradation experiments showed that ceramide post-translationally stabilizes the beta-secretase P56817 . Taken together , these data indicate that the lipid second messenger ceramide , which is elevated in the brains of Alzheimer 's disease patients , increases the half-life of P56817 and thereby promotes Abeta biogenesis . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Deletion of the prostaglandin E2 EP2 receptor reduces oxidative damage and amyloid burden in a model of Alzheimer 's disease . Epidemiological studies demonstrate that chronic use of nonsteroidal anti-inflammatory drugs ( NSAIDs ) in normal aging populations reduces the risk of developing Alzheimer 's disease ( AD ) . NSAIDs inhibit the enzymatic activity of cyclooxygenase-1 ( P23219 ) and inducible P35354 , which catalyze the first committed step in the synthesis of prostaglandins . These studies implicate P36551 -mediated inflammation as an early and potentially reversible preclinical event ; however , the mechanism by which P36551 activity promotes development of AD has not been determined . Recent studies implicate the prostaglandin E2 ( DB00917 ) E prostanoid subtype 2 ( EP2 ) receptor in the development of the innate immune response in brain . Here , we report that deletion of the DB00917 EP2 receptor in the APPSwe-PS1DeltaE9 model of familial AD results in marked reductions in lipid peroxidation in aging mice . This reduction in oxidative stress is associated with significant decreases in levels of amyloid-beta ( Abeta ) 40 and 42 peptides and amyloid deposition . Aged APPSwe-PS1DeltaE9 mice lacking the EP2 receptor harbor lower levels of beta C-terminal fragments , the product of beta-site P05067 cleaving enzyme ( P56817 ) processing of amyloid precursor protein . Increases in P56817 processing have been demonstrated in models of aging and AD and after oxidative stress . Our results indicate that DB00917 signaling via the EP2 receptor promotes age-dependent oxidative damage and increased Abeta peptide burden in this model of AD , possibly via effects on P56817 activity . Our findings identify EP2 receptor signaling as a novel proinflammatory and proamyloidogenic pathway in this model of AD , and suggest a rationale for development of therapeutics targeting the EP2 receptor in neuroinflammatory diseases such as AD . 5- Q9H205 - and P28335 -antagonist properties of cyamemazine : significance for its clinical anxiolytic activity . RATIONALE : DB09000 is a neuroleptic compound which possesses anxiolytic properties in humans . On the other hand , 5- Q9H205 - and P28335 -receptors have been implicated in anxiety disorders and a previous binding study has shown that cyamemazine possesses high affinity for both serotonin receptor types . OBJECTIVE : The present study was undertaken to establish whether cyamemazine antagonizes 5- Q9H205 - and/or P28335 -mediated responses , and whether it compares with reference compounds . METHODS : DB09000 was tested for its ability to antagonize : ( i ) 5- Q9H205 -dependent contraction of the isolated guinea-pig ileum and bradycardic responses in the rat and ( ii ) P28335 -dependent phospholipase C ( P98160 ) stimulation in rat brain membranes . RESULTS : In isolated guinea-pig ileum , cyamemazine potently and competitively antagonized 5-HT-dependent contractions ( pA2 = 7.52 +/- 0.08 ; n = 5 ) . In this test , cyamemazine was 5-7 times more potent ( pIC50 = 6.75 +/- 0.13 ) than tropisetron ( pIC50 = 6.02 +/- 0.04 ) . In rats , cyamemazine i.v. antagonized 5-HT-dependent bradycardic responses with ID50 % = 3.2 +/- 1.5 mg/kg ( n = 4 ) . Finally , in rat brain membranes cyamemazine antagonized P28335 -dependent P98160 stimulation with Ki = 424 nM ( mianserin exhibits a Ki = 113 nM ) . CONCLUSIONS : DB09000 behaves as an antagonist at both 5- Q9H205 - and P28335 -receptors , which compares well with reference compounds . These 5- Q9H205 - and P28335 -antagonistic actions of cyamemazine can be involved , at least in part , in its beneficial therapeutic actions in anxiety disorders . Association study between Alzheimer 's disease and genes involved in Abeta biosynthesis , aggregation and degradation : suggestive results with P56817 . BACKGROUND : Amyloid beta-peptide ( Abeta ) biosynthesis , aggregation and degradation constitute three important steps to consider in the study of pathological mechanisms involved in Alzheimer 's disease ( AD ) . Several proteins have been suggested as involved in each of these processes : proteolytic cleavage of the amyloid precursor protein by the beta-site P05067 cleaving enzyme ( P56817 ) , increased amyloid fibril formation by the activity of the acetylcholinesterase ( P22303 gene ) , and degradation of Abeta aggregates by the plasmin system have been exhaustively documented . METHODS : A case-control design was used to evaluate the possible association between candidate genes involved in these three processes and AD . We analysed three polymorphisms located at the P56817 gene , one polymorphism at the P22303 gene , and two variants located at the tissue plasminogen activator and plasminogen activator inhibitor-1 ( genes TPA and P05121 - 1 , respectively ) , both part of the plasmin system . RESULTS : We found an association between P56817 exon 5 GG genotype and AD ( age-and gender-adjusted odds ratio = 2.14 , P =0.014 ) . Although a similar association was reported previously by Nowotny and collaborators only in subjects carrying the epsilon4-allele of the apolipoprotein E gene ( P02649 ) , we did not detect this effect . However,when we combined our results with those previously reported , a clear increase of the risk to develop AD appeared in subjects carrying both the P56817 exon 5 GG genotype and the P02649 epsilon4-allele ( crude OR = 2.2 , P = 0.004 ) . CONCLUSION : These data suggest a possible genetic relation between P56817 and AD . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Statins exhibit anticancer effects through modifications of the pAkt signaling pathway . Statins are cholesterol lowering drugs that exhibit antitumor effects in several in vitro and in vivo models , and epidemiological studies indicate that statins prevent cancer . However , the molecular mechanism underlying the effects of statins still needs to be elucidated . We previously demonstrated that single doses of different statins rapidly affect Akt signaling via the purinergic receptor Q99572 . In particular , statins down-regulated nuclear pAkt . Here , we report that long-term treatment of A549 cells with high concentrations of statins ( 15-75 µM ) selects cell sub-populations exhibiting altered P2X receptor expression , signs of increased P60484 activity , enhanced Q6ZVD8 , decreased PI3K p110β and inhibited downstream pAkt signaling . Furthermore , the nuclear accumulation of pAkt in response to insulin was inhibited in selected cells . Statin-selected cells displayed reduced proliferation rate and were more vulnerable to etoposide- and 5-fluorouracil-elicited cytotoxic effects . The stability of a selected phenotype ( 50 µM ) was tested for three weeks in the absence of statins . This resulted in a reversal of some , but not all alterations . Importantly , the truncated nuclear insulin response was retained . We conclude that long-term treatment with high doses of statins selects cells exhibiting stable alterations in insulin-Akt signaling and which are vulnerable to DNA damage . Our studies strengthen the hypothesis that an altered Akt signaling has a role in chemopreventive effects of statins . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Calpain activation promotes P56817 expression , amyloid precursor protein processing , and amyloid plaque formation in a transgenic mouse model of Alzheimer disease . Abnormal activation of calpain is implicated in synaptic dysfunction and participates in neuronal death in Alzheimer disease ( AD ) and other neurological disorders . Pharmacological inhibition of calpain has been shown to improve memory and synaptic transmission in the mouse model of AD . However , the role and mechanism of calpain in AD progression remain elusive . Here we demonstrate a role of calpain in the neuropathology in amyloid precursor protein ( P05067 ) and presenilin 1 ( P49768 ) double-transgenic mice , an established mouse model of AD . We found that overexpression of endogenous calpain inhibitor calpastatin ( CAST ) under the control of the calcium/calmodulin-dependent protein kinase II promoter in P05067 / P49768 mice caused a remarkable decrease of amyloid plaque burdens and prevented Tau phosphorylation and the loss of synapses . Furthermore , CAST overexpression prevented the decrease in the phosphorylation of the memory-related molecules CREB and P29323 in the brain of P05067 / P49768 mice and improved spatial learning and memory . Interestingly , treatment of cultured primary neurons with amyloid-beta ( Abeta ) peptides caused an increase in the level of beta-site P05067 -cleaving enzyme 1 ( P56817 ) , the key enzyme responsible for P05067 processing and Abeta production . This effect was inhibited by CAST overexpression . Consistently , overexpression of calpain in heterologous P05067 expressing cells up-regulated the level of P56817 and increased Abeta production . Finally , CAST transgene prevented the increase of P56817 in P05067 / P49768 mice . Thus , calpain activation plays an important role in P05067 processing and plaque formation , probably by regulating the expression of P56817 . An aging pathway controls the TrkA to p75NTR receptor switch and amyloid beta-peptide generation . Aging of the brain is characterized by marked changes in the expression levels of the neurotrophin receptors , TrkA and p75(NTR) . An expression pattern in which TrkA predominates in younger animals switches to one in which p75(NTR) predominates in older animals . This TrkA-to-p75(NTR) switch is accompanied by activation of the second messenger ceramide , stabilization of beta-site amyloid precursor protein-cleaving enzyme-1 ( P56817 ) , and increased production of amyloid beta-peptide ( Abeta ) . Here , we show that the insulin-like growth factor-1 receptor ( IGF1-R ) , the common regulator of lifespan and age-related events in many different organisms , is responsible for the TrkA-to-p75(NTR) switch in both human neuroblastoma cell lines and primary neurons from mouse brain . The signaling pathway that controls the level of TrkA and p75(NTR) downstream of the IGF1-R requires Q9Y4H2 , PIP3/Akt , and is under the control of P60484 and Q8TCB0 , the short isoform of p53 . We also show that hyperactivation of IGF1-R signaling in Q8TCB0 transgenic animals , which show an accelerated form of aging , is characterized by early TrkA-to-p75(NTR) switch and increased production of Abeta in the brain . Prolonged effects of tumor necrosis factor-alpha on anterior pituitary hormone release . We examined the chronic ( 72 h ) effects of 30 ng/ml recombinant murine tumor necrosis factor ( P01375 ) -alpha on release of immunoreactive growth hormone ( GH ) , prolactin ( PRL ) , thyrotropin ( DB00024 ) , and DB00024 glycosylation , as assessed by lectin binding , in cultured rat anterior pituitary cells . In cultured cells from adult female rats , P01375 significantly suppressed basal and GH-releasing hormone ( P01148 ) -stimulated GH release . P01375 also suppressed basal PRL release and completely abolished the PRL response to TRH ( 0.1-10 nM ) . Whereas P01375 reduced basal DB00024 release , it significantly enhanced the maximal DB00024 response to TRH . P01375 did not affect the concanavalin A and lentil lectin binding of DB00024 accumulated in the medium during the 4-day culture , but significantly decreased the lentil lectin binding of DB00024 released in response to acute TRH stimulation . P01375 significantly enhanced the inhibitory effect of somatostatin on stimulated PRL release , but not on GH or DB00024 release . Compared to cell cultures from adult female rats , in anterior pituitary cell cultures from 12-day-old rats the effects of prolonged exposure to P01375 on hormone release were diminished or absent . Pituitary hormone release was unaffected by acute ( 3 h ) exposure to P01375 . These results demonstrate a direct effect of P01375 on anterior pituitary hormone release , which is cell-type specific and age dependent . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP .

MH_train_1056

MH_train_1056

MH_train_1056

interacts_with DB00482?

multiple_choice

Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Are rofecoxib and celecoxib safer NSAIDS ? NSAIDs work by inhibiting the enzyme cyclo-oxygenase ( P36551 ) , responsible for prostaglandin synthesis . This enzyme exists in two isoforms , P23219 and P35354 . Inhibition of P23219 is thought to be the main cause of the gastrointestinal unwanted effects of NSAIDs , whilst inhibition of P35354 results in anti-inflammatory effects . [ symbol : see text ] DB00533 ( Vioxx -- MSD ) and [ symbol : see text ] celecoxib ( DB00482 -- Searle ) have been developed as selective inhibitors of P35354 . DB00533 is licensed for the symptomatic treatment of osteoarthritis , but not for rheumatoid arthritis . The manufacturer claims that " in clinical studies rofecoxib inhibits P35354 but not P23219 " , has " the power of high-dose NSAIDs -- diclofenac and ibuprofen " and " superior GI safety profile compared to conventional NSAIDs " . Celecoxib is licensed for symptom relief in osteoarthritis and rheumatoid arthritis . The manufacturer claims that celecoxib has " comparable efficacy and superior GI tolerability when compared to diclofenac or naproxen " . Here , we review rofecoxib and celecoxib and consider whether they are safer than conventional NSAIDs . P35354 inhibitors : better than traditional NSAIDs ? Vioxx and DB00482 may be no less risky than NSAIDs . Purinergic P47900 receptor signaling mediates wound stimuli-induced cyclooxygenase-2 expression in intestinal subepithelial myofibroblasts . Intestinal subepithelial myofibroblasts ( ISMFs ) are crucial for barrier formation against inflammatory stimuli . Physical injury induces cyclooxygenase-2 ( P35354 ) expression , which accelerates wound healing by ISMFs . However , the mechanism of P35354 induction remains unclear . Physically damaged cells release DB00171 . Here , we investigate the role of DB00171 -purinergic signaling in wound-induced P35354 induction in ISMFs . By 24h post-injury , bovine ISMFs had migrated to and closed the wounded area . A P36551 inhibitor , indomethacin or a purinergic P2 receptor antagonist , suramin , inhibited wound healing . However , additional treatment with indomethacin did not influence wound healing in suramin-treated ISMFs . RT-PCR showed an increase in P35354 mRNA expression 2h post-injury , which was inhibited by suramin . These results suggest that DB00171 mediates wound-induced P35354 elevation . We next assessed the contribution of various purinergic receptors in P35354 induction . An DB00171 analog , ATPγS and a purinergic P47900 , 11-13 receptors agonist , ADP , were among the agents tested which increased P35354 expression . ATPγS-induced P35354 mRNA expression was suppressed by suramin or a purinergic P2Xs , P47900 , 4 , 6 , and 13 receptors antagonist , PPADS . These data suggest the involvement of Gq-coupled purinergic P47900 receptor or Gi-coupled purinergic Q9BPV8 receptor in P35354 induction . U73122 , an inhibitor of phospholipase C , which is a downstream signal of Gq protein , showed suppression of P35354 mRNA expression . However , pertussis toxin , a Gi inhibitor , did not show suppression . We also revealed that inhibitors of p38 MAPK and PKC inhibited ATPγS-induced P35354 mRNA expression . Collectively , purinergic P47900 receptor signaling mediates wound-induced P35354 expression through p38 MAPK and PKC pathways in ISMFs . Distinct effects of inflammation on gliosis , osmohomeostasis , and vascular integrity during amyloid beta-induced retinal degeneration . In normal retinas , amyloid-β ( Aβ ) accumulates in the subretinal space , at the interface of the retinal pigment epithelium , and the photoreceptor outer segments . However , the molecular and cellular effects of subretinal Aβ remain inadequately elucidated . We previously showed that subretinal injection of Aβ(1-42) induces retinal inflammation , followed by photoreceptor cell death . The retinal Müller glial ( RMG ) cells , which are the principal retinal glial cells , are metabolically coupled to photoreceptors . Their role in the maintenance of retinal water/potassium and glutamate homeostasis makes them important players in photoreceptor survival . This study investigated the effects of subretinal Aβ(1-42) on RMG cells and of Aβ(1-42)-induced inflammation on retinal homeostasis . RMG cell gliosis ( upregulation of P14136 , vimentin , and nestin ) on day 1 postinjection and a proinflammatory phenotype were the first signs of retinal alteration induced by Aβ(1-42) . On day 3 , we detected modifications in the protein expression patterns of cyclooxygenase 2 ( P35354 ) , glutamine synthetase ( GS ) , Kir4.1 [ the inwardly rectifying potassium ( Kir ) channel ] , and aquaporin ( AQP ) -4 water channels in RMG cells and of the photoreceptor-associated P29972 . The integrity of the blood-retina barrier was compromised and retinal edema developed . Aβ(1-42) induced endoplasmic reticulum stress associated with sustained upregulation of the proapoptotic factors of the unfolded protein response and persistent photoreceptor apoptosis . Indomethacin treatment decreased inflammation and reversed the Aβ(1-42)-induced gliosis and modifications in the expression patterns of P35354 , Kir4.1 , and P29972 , but not of P55087 or GS . Nor did it improve edema . Our study pinpoints the adaptive response to Aβ of specific RMG cell functions . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . The new P35354 inhibitors : rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) . Anti-angiogenic effects of Hypericin-photodynamic therapy in combination with DB00482 in the treatment of human nasopharyngeal carcinoma . Photodynamic therapy ( PDT ) is being investigated as an alternative treatment modality in cancer treatment . It has been shown to induce tumor hypoxia and upregulation of cyclooxygenase-2 ( P35354 ) and vascular endothelial growth factor ( P15692 ) . The objective of this study was to improve in vivo tumor growth control of nasopharyngeal carcinoma ( NPC ) , treated at a subcurative dosage by using a combination of Hypericin-PDT and P35354 inhibitor , DB00482 ( CX ) . The effect of an initial CX dose at 6- and 24-h post-PDT was investigated simultaneously . It was observed that hypoxic NPC/CNE2 cells upregulate both P35354 and P15692 A genes in vitro . In vivo studies , down-regulation of P35354 and hypoxia inducible factor-1alpha ( HIF-1alpha ) genes at 24-h post-PDT and bulk tumor ablation at 48-h post-PDT was observed . However , 24-28 days later regrowth was observed . In a combination treatment , 1st CX dose at 6-h post-PDT had the highest tumor control in which tumors were < or=0.52 cm(3) ( 64.29 % , P < 0.05 ) . However , the tumors administered with a initial dose of CX at 24-h post-PDT had no tumor control . Co-suppression of P35354 , HIF-1alpha and P15692 A genes were observed in tumors with tumor control . Mice without tumor control that were subjected to therapy had increased human P15692 in serum compared to Hypericin-PDT mice . Thus , suggesting that the time of initial administration of CX post-PDT is an important factor for effective tumor control . Influence of polymers on the bioavailability of microencapsulated celecoxib . Celecoxib , a selective P35354 inhibitor , primarily used in treatment of osteoarthritis , rheumatoid arthritis and acute pain was encapsulated in microparticles composed of various polyesters , polymethacrylates or cellulose derivatives used alone or blended . The influence of polymers on microparticle mean diameter , encapsulation efficiency and in vitro and in vivo celecoxib release was investigated . Microparticles were in the size range 11-37 microm . Encapsulation efficiency was optimal due to poor aqueous solubility of celecoxib . Considering in vitro release , microparticles could be divided into drug delivery systems with fast and slow release profiles . Microparticles prepared with poly-epsilon-caprolactone , Eudragit RS and low viscosity ethylcellulose , together with physical mixture of celecoxib with lactose and DB00482 , were tested in vivo . Relative bioavailability of celecoxib was below 20 % in all cases and was probably the consequence of a slow in vivo release of celecoxib from microparticles or low wettability in the case of DB00482 and physical mixture . P35354 inhibitors in glioma therapy . The cyclooxygenase enzyme is a prostaglandin synthase that has two isoforms , cyclooxygenase-1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) . Upregulation of the P35354 isoform in many cancers has led to investigation regarding the potential role of this enzyme in oncogenesis . The Food and Drug Administration has approved celecoxib ( DB00482 ) for patients with familial adenomatous polyposis ( FAP ) based on a study that showed a reduction in the number of polyps in such patients when treated with high doses of this drug . We are investigating the potential role of P35354 inhibitors in the treatment of recurrent malignant gliomas in combination with retinoids . In this article the rationale for using this combination therapy in patients with malignant gliomas is presented . Regional expression and role of cyclooxygenase-2 following experimental traumatic brain injury . Prostaglandins , potent mediators of inflammation , are generated from arachidonic acid ( AA ) via the action of cyclooxygenase-1 and -2 ( P23219 and P35354 ) . In this study , we report that lateral cortical impact injury in rats significantly increases P35354 protein levels both in the cortex surrounding the injury site and the ipsilateral hippocampus . P35354 protein level was elevated as early as 3 h postinjury and persisted for up to 3 days . Increases in immunoreactivity were detected not only in the adjacent cortex and hippocampus , but were also observed in the contralateral cortex and hippocampus , the ipsilateral piriform cortex and the ipsilateral amygdaloid complex . P35354 immunoreactive cells appear morphologically normal and do not present any of the characteristic features of apoptosis . Double immunostaining experiments using either a neuron-specific or an astroglial-specific marker show that the expression of P35354 is localized almost exclusively in neuronal cells . Administration of the P35354 inhibitor 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfona mide ( celecoxib , marketed as DB00482 ) worsens motor , but not cognitive , performance , suggesting that P35354 induction following traumatic brain injury may play a protective role . Modulation of pulmonary leukotriene B4 production by cyclooxygenase-2 inhibitors and lipopolysaccharide . PURPOSE : Emerging data continue to link carcinogenesis to inflammatory events involving the eicosanoid metabolic pathways . We therefore evaluated the effects of cyclooxygenase ( P36551 ) -2 inhibition on leukotriene ( LT ) B(4) synthesis in the lungs of active smokers , as part of a pilot lung cancer chemoprevention study with celecoxib ( DB00482 ) , an oral P35354 inhibitor . EXPERIMENTAL DESIGN : Bronchoalveolar lavage was performed before celecoxib treatment and after 1 month of celecoxib treatment to recover alveolar macrophages ( AMs ) and lining fluid for study . After harvest , AMs were immediately stimulated in vitro with the calcium ionophore A23187 . AMs obtained from smokers before treatment and from ex-smoker control subjects were also cultured overnight with SC58236 , a selective P35354 inhibitor , with or without lipopolysaccharide stimulation . RESULTS : Treatment with oral celecoxib only modestly increased Q06643 (4) levels in bronchoalveolar lavage , without increasing the mRNA transcription of P09917 ( 5- P28300 ) or 5- P28300 -activating protein in AMs , whereas the acute calcium ionophore-stimulated Q06643 (4) production from smokers ' AMs was markedly increased by 10.6-fold . In addition , smokers ' AMs were twice as responsive in producing Q06643 (4) when exposed to lipopolysaccharide compared with ex-smokers ' AMs . Concomitant P35354 inhibition with SC58236 , however , did not significantly impact these changes , whereas the 5- P28300 inhibitor Zileuton blocked the generation of Q06643 (4) in a dose-responsive manner . Finally , cycloheximide increased the production of Q06643 (4) under all conditions , suggesting a shunting phenomenon and/or the presence of pathway inhibitors . CONCLUSIONS : Our findings suggest that whereas oral celecoxib is capable of modulating Q06643 (4) production in the lung microenvironment , under physiologic conditions , this effect is probably not functionally significant . P35354 inhibitors : a story of greed , deception and death . In 1999 , drug manufacturers introduced a class of NSAIDs called P35354 inhibitors or coxibs . The drugs were avidly promoted directly to the consumers and became bestsellers from the start . Arthritis sufferers were eager to take medications that eased joint pain with less risk of causing gastrointestinal pain , bleeding and other side-effects . In the year after their introduction , doctors wrote over 100 million prescriptions for celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . DB00482 is the sixth best-selling drug , with sales of more than US $ 4 billion since its debut in 1999 . Vioxx had sales of US $ 2.6 billion in 2001 . However , the coxibs increase the risk of heart attacks and strokes , and their price , in the USA , is obscene . The manufacturers faced a possibly complicit , toothless and bloodless FDA , and used every maneuvering to fleece the patients . We must now reflect on attitudes that we thought only belong to the tobacco industry . Fortunately , safe and active alternatives exist . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Plasma levels of vascular endothelial growth factor during and after radiotherapy in combination with celecoxib in patients with advanced head and neck cancer . DB00482 and radiotherapy in advanced head and neck cancer . This phase I dose-escalation study seeks to determine the phase II recommended dose of cyclooxygenase type 2 ( P35354 ) inhibitor in patients with locally advanced squamous cell head and neck ( H & N ) cancer , treated with accelerated radiotherapy . Anti-vasculogenic effect of this treatment on serum vascular endothelial growth factor ( P15692 ) is examined . Patients were irradiated with curative intent ( 72Gy in 6weeks ) . Celecoxib was administered throughout the radiotherapy course . Serum P15692 level were tested during radiotherapy and in follow-up . Tumor specimens were stained to quantify the P35354 expression . Thirty-two patients completed the treatment . The dose of celecoxib was escalated ( 200 , 400 and 800mg bid , then de-escalated to 600mg bid ) . The acute toxicity related to the treatment in the first and second cohort did not reach grade III ; in the third cohort three patients had grade III radiation toxicity and one had celecoxib-related toxicity . In the last fourth cohort the toxicity was acceptable . Significant P15692 level drop ( p=0.011 ) was found between radiation day 1 and post-treatment visit . Significant decrease ( p=0.022 ) of the P15692 level was shown in patients with high P35354 expression in the tumor . Phase II recommended dose of celecoxib combined with accelerated radiotherapy in advanced H & N cancer was 600mg bid . A significant decrease of the post-treatment serum P15692 level compared to the initial level was noticed only in patients with high P35354 expression in tumors . P35354 inhibition in clinical cancer prevention . Colorectal cancer is an excellent model for studying cancer prevention by means of secondary ( e.g. , polypectomy to remove a precursor adenoma ) and primary ( chemoprevention ) strategies . Evidence has shown that regular users of aspirin and other nonsteroidal anti-inflammatory drugs ( NSAIDs ) have a reduction in risk of colorectal cancer . A possible mechanism of this benefit is decreased prostaglandin production , which is achieved through inhibition of cyclooxygenase ( P36551 ) activity , and possibly other pathways . Two isoforms of P36551 -- P23219 and P35354 -- have been identified . P35354 is expressed in colorectal adenomas and carcinomas , both in humans and rodents . Inhibition of P35354 has been shown to decrease the incidence of carcinogen-induced neoplasia in rats and to lower the incidence of adenomas in murine models . Several P35354 inhibitors , with the potential for less toxicity than that associated with traditional NSAIDs , are under development . This paper reviews potential chemoprevention of colorectal cancer using P35354 inhibitors in patients at increased risk , e.g. , patients with familial adenomatous polyposis , hereditary nonpolyposis colorectal cancer , and sporadic adenomas . Included are the rationale for use of such agents , results of a study showing a significant reduction in adenoma burden in familial adenomatous polyposis patients who received the selective P35354 inhibitor celecoxib ( DB00482 ) , and the design of other ongoing or planned clinical trials . Celecoxib : a novel treatment for lung cancer . Lung cancer is by far the leading cause of cancer-related deaths . Overall survival is poor and has not improved substantially over the last 50 years . Therefore , it is clear that novel and more effective treatments are needed to improve the outcome of therapy . Recent attention has been drawn to the role of cyclooxygenase ( P36551 ) -2 in the pathogenesis of cancer , and it has been considered as an attractive target for therapeutic and chemopreventive strategies in lung cancer patients . Celecoxib ( DB00482 ) , Pfizer ) , a selective P35354 inhibitor and potent anti-inflammatory agent , has been approved for the treatment of osteoarthritis and rheumatoid arthritis . This orally administered agent is generally well tolerated and has almost no gastrointestinal or renal toxicity . Phase II clinical trials suggest that P35354 inhibition by celecoxib would enhance response to cytotoxic chemotherapy or radiation therapy through interference with cellular proliferation and tumor angiogenic processes , promotion of apoptosis and immune surveillance , or other possible mechanisms . Celecoxib has shown promising antitumor efficacy in lung cancer and a large variety of solid tumors that rely on P35354 -related mechanisms for growth and survival . This article reviews the profile of celecoxib and evidence supporting its role in the therapy of lung cancer . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . Current application of selective P35354 inhibitors in cancer prevention and treatment . The multistep process of carcinogenesis , which can take many years , provides many opportunities for intervention to inhibit disease progression . Effective chemoprevention agents may reduce the risk of cancer by inhibiting the initiation stage of carcinoma through induction of apoptosis or DNA repair in cells harboring mutations , or they may act to prevent promotion of tumor growth . Similarly , chemoprevention may entail blocking cancer progression to an invasive phenotype . Over the past decade , in vitro , preclinical , and clinical data have supported the hypothesis that cyclooxygenase ( P36551 ) -2 plays a central role in oncogenesis and that treatment with P35354 inhibitors offers an effective chemoprevention strategy , as exemplified by the activity of celecoxib ( DB00482 ) in familial adenomatous polyposis . These P35354 data have contributed to initiation of clinical trials testing P35354 inhibitors for the chemoprevention of a wide variety of cancers that overexpress P35354 . Characterization of the effects of antiangiogenic agents on tumor pathophysiology . A variety of strategies have been proposed to control tumor growth and metastasis by inhibiting tumor angiogenesis . To optimally combine such antiangiogenic approaches with conventional therapy , improved methods are needed to characterize the underlying pathophysiologic changes . The objective of the current work was to demonstrate the utility of a combination of recently developed immunohistochemical and image analysis techniques in quantitating changes in tumor vasculature and hypoxia . Murine MCa-35 mammary carcinomas were frozen after administration of two P35354 inhibitors : meloxicam and celecoxib ( DB00482 ) . Total blood vessels were visualized using anti-CD31 staining , perfused vessels by intravenous injection of DiOC7 , and tumor hypoxia by EF5 uptake . Although both agents produced similar reductions in tumor volume compared with untreated tumors , varied effects on tumor vasculature and hypoxia were noted . Meloxicam reduced total vessel numbers significantly , whereas celecoxib had no effect . Both drugs substantially increased perfused vessel densities . Although mean hypoxic marker uptake was unchanged from matched controls , intratumor EF5 heterogeneities were significantly different between drugs . The results suggest that P35354 inhibitors can have varying effects on tumor pathophysiology . Successful use of these drugs to enhance radiation response will likely require optimization of drug choice , dose schedule , and direct physiologic monitoring . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . P14210 induces anoikis resistance by up-regulation of cyclooxygenase-2 expression in uterine endometrial cancer cells . P35354 ( P35354 ) has been implicated in the promotion of carcinogenesis . Although the role of P35354 in endometrial cancer remains unclear , recent experiments suggest that P35354 antagonizes cell apoptosis , increases the invasiveness of malignant cells , and promotes angiogenesis . P14210 ( P14210 ) is a mesenchymal-derived cytokine and the interaction between P14210 and its tyrosine kinase receptor , c- DB00134 proto-oncogene , is associated with tumor progression and metastasis . To investigate the molecular mechanism of P14210 -induced anoikis resistance , we analyzed the signal transduction and P35354 expression in endometrial cancer cells . Here , we show i ) the expression of P35354 protein significantly increased in a dose-dependent manner after P14210 stimulation in endometrial cancer cell lines ( O14777 -IB and RL95-2 ) , reaching 200-270 % stimulation at the highest doses of P14210 tested ( 40 ng/ml ) ; ii ) flow cytometry and TUNEL analyses revealed that P14210 significantly inhibited anoikis of RL95-2 cells ; iii ) phosphatidylinositol 3-kinase ( PI3K ) inhibitor ( LY294002 ) , but not mitogen-activated protein kinase/ P29323 kinase ( MEK ) inhibitor ( PD98059 ) , specifically blocked P14210 -mediated anoikis resistance in RL95-2 cells ; and iv ) P35354 inhibitor , Meloxicam , abrogated P14210 -mediated anoikis resistance . Our data suggest that P14210 induces anoikis resistance in endometrial cancer cells possibly through PI3K/Akt pathway-dependent up-regulation of P35354 expression . Lipoperoxidation and cyclooxygenases 1 and 2 inhibitory compounds from Iryanthera juruensis . Plants from Iryanthera genus have been traditionally used as food supplements by South American Indians . The MeOH extract of leaves of Iryanthera juruensis , one of the plants endemic to the Amazon region and consumed in Brazil , and the hexane extract from its seeds inhibited lipid peroxidation ( P22079 ) and cyclooxygenase ( P23219 and -2 ) ) enzymes in in vitro assays . Further analyses of these extracts yielded 5-deoxyflavones ( 1-5 ) from the leaf extract and sargachromenol ( 6 ) , sargaquinoic acid ( 7 ) , a novel juruenolic acid ( 8 ) , omega-arylalkanoic acids ( 9a-c ) , and the lignan guaiacin ( 10 ) from the seed extract . Compounds 3-5 inhibited P22079 by 86 % , 77 % , and 88 % at 10 ppm , respectively , and compounds 6 and 9a-c showed inhibition at 76 % and 78 % at 100 ppm , respectively . However , compounds 7 and 8 were inactive and lignan 10 exhibited P22079 inhibitory activity by 99 % at 100 ppm compared to commercial antioxidants butylated hydroxyanisole ( BHA ) , butylated hydroxytoluene ( BHT ) , and vitamin E . The flavones 1-5 also inhibited P23219 and -2 enzymes by 50-65 % at 100 ppm . DB04088 showed high but nonselective inhibition of P23219 and P35354 enzymes , when compared to aspirin and DB00482 , a nonsteroidal anti-inflammatory drug ( NSAID ) . Compounds 7 and 10 inhibited P23219 by 60 % and 65 % and P35354 by 37 % and 18 % , respectively , whereas compounds 8 and 9a-c showed little or no activity against these enzymes . Correcting human mitochondrial mutations with targeted RNA import . Mutations in the human mitochondrial genome are implicated in neuromuscular diseases , metabolic defects , and aging . An efficient and simple mechanism for neutralizing deleterious mitochondrial DNA ( mtDNA ) alterations has unfortunately remained elusive . Here , we report that a 20-ribonucleotide stem-loop sequence from the H1 RNA , the RNA component of the human RNase P enzyme , appended to a nonimported RNA directs the import of the resultant RNA fusion transcript into human mitochondria . The methodology is effective for both noncoding RNAs , such as tRNAs , and mRNAs . The RNA import component , polynucleotide phosphorylase ( Q8TCS8 ) , facilitates transfer of this hybrid RNA into the mitochondrial matrix . In addition , nucleus-encoded mRNAs for mitochondrial proteins , such as the mRNA of human mitochondrial ribosomal protein P28222 ( O15235 ) , contain regulatory sequences in their 3'-untranslated region ( UTR ) that confers localization to the mitochondrial outer membrane , which is postulated to aid in protein translocation after translation . We show that for some mitochondrial-encoded transcripts , such as P35354 , a 3'-UTR localization sequence is not required for mRNA import , whereas for corrective mitochondrial-encoded tRNAs , appending the 3'-UTR localization sequence was essential for efficient fusion-transcript translocation into mitochondria . In vivo , functional defects in mitochondrial RNA ( mtRNA ) translation and cell respiration were reversed in two human disease lines . Thus , this study indicates that a wide range of RNAs can be targeted to mitochondria by appending a targeting sequence that interacts with Q8TCS8 , with or without a mitochondrial localization sequence , providing an exciting , general approach for overcoming mitochondrial genetic disorders . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . Inflammation induces mitochondrial dysfunction and dopaminergic neurodegeneration in the nigrostriatal system . Evidence suggests that chronic inflammation , mitochondrial dysfunction , and oxidative stress play significant and perhaps synergistic roles in Parkinson 's disease ( PD ) , where the primary pathology is significant loss of the dopaminergic neurons in the substantia nigra . The use of anti-inflammatory drugs for PD treatment has been proposed , and inhibition of cyclo-oxygenase-2 ( P35354 ) or activation of peroxisome proliferator-activated receptor gamma ( P37231 ) yields neuroprotection in MPTP-induced PD . Lipopolysaccharide ( LPS ) induces inflammation-driven dopaminergic neurodegeneration . We tested the hypothesis that celecoxib ( DB00482 , P35354 inhibitor ) or pioglitazone ( Actos , P37231 agonist ) will reduce the LPS-induced inflammatory response , spare mitochondrial bioenergetics , and improve nigral dopaminergic neuronal survival . Rats were treated with vehicle , celecoxib , or pioglitazone and were intrastriatally injected with LPS . Inflammation , mitochondrial dysfunction , oxidative stress , decreased dopamine , and nigral dopaminergic neuronal loss were observed post-LPS . Celecoxib and pioglitazone provided neuroprotective properties by decreasing inflammation and restoring mitochondrial function . Pioglitazone also attenuated oxidative stress and partially restored striatal dopamine as well as demonstrated dopaminergic neuroprotection and reduced nigral microglial activation . In summary , intrastriatal LPS served as a model for inflammation-induced dopaminergic neurodegeneration , anti-inflammatory drugs provided protective properties , and pioglitazone or celecoxib may have therapeutic potential for the treatment of neuro-inflammation and PD . Celecoxib exhibits the greatest potency amongst cyclooxygenase ( P36551 ) inhibitors for growth inhibition of P35354 -negative hematopoietic and epithelial cell lines . P35354 ( P35354 ) is an important cellular target for both therapy and/or prevention of inflammatory disorders and cancer . The advent of selective P35354 inhibitors now allows a more precise and safer treatment approach . The screening of an array of cancer cell lines for growth inhibitory effects of P35354 -selective and -nonselective inhibitors , including celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) , produced two unanticipated findings . Firstly , the antiproliferative effects of celecoxib were noted to be of very similar magnitude for both hematopoietic and epithelial cancer cell lines . Most hematopoietic cell lines had no detectable P35354 expression by reverse transcription-PCR , and none expressed P35354 protein . In addition , P35354 -negative epithelial lines were found to have IC50s for celecoxib that were very similar to their P35354 + counterparts . Thus , important antiproliferative effects were observed that were independent of both the cell lineage and P35354 status . Secondly , it was also observed that P35354 inhibitor drugs , celecoxib and rofecoxib , with similar P35354 -selectivity and clinical efficacy for inflammatory indications , differed significantly in their in vitro antiproliferative effects on cancer cell lines . IC50s of 35-65 microM were observed for celecoxib across this entire panel of cell lines . Finally , no difference in the mode or degree of cytotoxicity was apparent between cell lines , because similar levels of apoptosis were observed in P35354 + and -negative cell lines after treatment with celecoxib , with correspondingly lower levels after rofecoxib treatment . These data are important in that they provide the first direct comparison of epithelial and hematopoietic cancer cell lines , as well as a direct comparison of the in vitro anticancer effects of the two clinically available P35354 inhibitors . DB00917 produced by lung cancer suppresses immune function through T-regulatory cells and can be blocked by the P35354 inhibitor DB00482 . Inhibition of delayed rectifier potassium channels and induction of arrhythmia : a novel effect of celecoxib and the mechanism underlying it . Selective inhibitors of cyclooxygenase-2 ( P35354 ) , such as rofecoxib ( Vioxx ) , celecoxib ( DB00482 ) , and valdecoxib ( Bextra ) , have been developed for treating arthritis and other musculoskeletal complaints . Selective inhibition of P35354 over P23219 results in preferential decrease in prostacyclin production over thromboxane A2 production , thus leading to less gastric effects than those seen with nonselective P36551 inhibitors such as acetylsalicylic acid ( aspirin ) . Here we show a novel effect of celecoxib via a mechanism that is independent of P35354 inhibition . The drug inhibited the delayed rectifier ( Kv2 ) potassium channels from Drosophila , rats , and humans and led to pronounced arrhythmia in Drosophila heart and arrhythmic beating of rat heart cells in culture . These effects occurred despite the genomic absence of cyclooxygenases in Drosophila and the failure of acetylsalicylic acid , a potent inhibitor of both P23219 and P35354 , to inhibit rat Kv2.1 channels . A genetically null mutant of Drosophila Shab ( Kv2 ) channels reproduced the cardiac effect of celecoxib , and the drug was unable to further enhance the effect of the mutation . These observations reveal an unanticipated effect of celecoxib on Drosophila hearts and on heart cells from rats , implicating the inhibition of Kv2 channels as the mechanism underlying this effect . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . A real time quantitative PCR analysis and correlation of P23219 and P35354 enzymes in inflamed dental pulps following administration of three different NSAIDs . Dental pain is encountered daily by clinicians . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) commonly used for pain management are traditionally cyclooxygenase-1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) inhibitors , and more recently selective P35354 inhibitors . This study was designed to identify and quantify P23219 and P35354 gene expression level in inflamed rat molar pulps after administration of three NSAIDs : DB00482 , Vioxx , and DB01050 . Fifty male Wistar rats had their first and second molar pulps exposed and sealed with Cavit for 4 days . Rats were randomly divided into the three drug groups and two control groups . RNA was isolated from the rat pulps . Real Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction assay , a relatively new PCR technique , was used to quantify P23219 and P35354 mRNA . Statistical analysis demonstrated no significant differences in P23219 and P35354 levels among the drug groups . However , Vioxx and DB01050 significantly reduced P35354 expression levels compared to inflamed ( positive control ) pulps ( p < 0.05 ) . Antitumor properties of dimethyl-celecoxib , a derivative of celecoxib that does not inhibit cyclooxygenase-2 : implications for glioma therapy . Celecoxib ( DB00482 ) appears to be unique among the class of selective P35354 inhibitors ( coxibs ) , because this particular compound exerts a second function that is independent of its celebrated ability to inhibit P35354 . This second function is the potential to inhibit cell proliferation and stimulate apoptotic cell death at much lower concentrations than any other coxibs . Intriguingly , these two functions are mediated by different moieties of the celecoxib molecule and can be separated . The author , as well as others , have generated and investigated analogs of celecoxib that retain only one of these two functions . One derivative , 2,5-dimethyl-celecoxib ( Q6UXB2 ) , which retains the antiproliferative and apoptosis-inducing function , but completely lacks the P35354 inhibitory activity , is able to mimic faithfully all of the numerous antitumor effects of celecoxib that have been investigated so far , including reduction of neovascularization and inhibition of experimental tumor growth in various in vivo tumor models . In view of the controversy that has recently arisen regarding the life-threatening side effects of this class of coxibs , it may be worthwhile to pursue further the potential benefits of drugs such as Q6UXB2 for anticancer therapy . Because Q6UXB2 is not a coxib yet potently maintains celecoxib 's antitumor potential , one may be inclined to speculate that this novel compound could potentially be advantageous in the management of P35354 -independent cancers . In this summary , the implications of recent findings with Q6UXB2 will be presented and discussed . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . P35354 independent effects of cyclooxygenase-2 inhibitors on oxidative stress and intracellular glutathione content in normal and malignant human B-cells . We recently reported that inhibition of P35354 ( Cox-2 ) reduced human B-CLL proliferation and survival . Herein , we investigated the mechanisms whereby small molecule Cox-2 selective inhibitors , SC-58125 ( a DB00482 analog ) and CAY10404 blunt survival of human B-cell lymphomas and chronic lymphocytic leukemia B-cells . SC-58125 and OSU03012 ( a DB00482 analog that lacks Cox-2 inhibitory activity ) both decreased intracellular glutathione ( DB00143 ) content in malignant human B-cells , as well as in Cox-2 deficient mouse B-cells . This new finding supports Cox-2 independent effects of SC-58125 . Interestingly , SC-58125 also significantly increased B-cell reactive oxygen species ( ROS ) production , suggesting that ROS are a pathway that reduces malignant cell survival . Addition of DB00143 ethyl ester protected B lymphomas from the increased mitochondrial membrane permeability and reduced survival induced by SC-58125 . Moreover , the SC-58125-mediated DB00143 depletion resulted in elevated steady-state levels of the glutamate cysteine ligase catalytic subunit mRNA and protein . These new findings of increased ROS and diminished DB00143 levels following SC-58125 exposure support novel mechanisms whereby a Cox-2 selective inhibitor reduces malignant B-cell survival . These observations also support the concept that certain Cox-2 selective inhibitors may have therapeutic value in combination with other drugs to kill malignant B lineage cells . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . Approach to angiogenesis inhibition based on cyclooxygenase-2 . Two cyclooxygenase ( P36551 ) isoforms have been identified : P23219 and P35354 . P23219 is the constitutively expressed form of the enzyme and is ubiquitous in its distribution . P35354 is inducible and is present in inflammatory foci , tumors , and neovasculature . Expression of P35354 appears to be important in tumor promotion , growth , and metastasis . It is up-regulated in a variety of premalignant disorders and malignancies . P36551 inhibitors have a major role in the treatment of inflammation and pain . Epidemiologic evidence in patients who take nonsteroidal anti-inflammatory drugs links P36551 inhibition with decreases in malignant esophageal , stomach , colon , lung , and breast tumors . Nonselective P36551 inhibitors have demonstrated efficacy in control of familial adenomatous polyposis , a disorder associated with the development of thousands of benign intestinal polyps . The selective P35354 inhibitor celecoxib ( DB00482 , Pharmacia ) has been shown to reduce the number of adenomatous colorectal polyps in familial adenomatous polyposis as an adjunct to usual care . Celecoxib has recently been approved for this indication and offers the potential for equivalent or greater efficacy than that seen with nonselective P36551 inhibitors but without the gastrointestinal mucosal toxicity and the inhibition of platelet function associated with those agents . Angiogenesis is a feature of both benign and malignant disease . Because P35354 is up-regulated in the neovasculature of the rheumatoid pannus and in malignant tumors and their surrounding stroma , selective P35354 inhibitors may be able to modify the progression of these disorders through the control of angiogenesis . A cyclooxygenase-2/prostaglandin E2 pathway augments activation-induced cytosine deaminase expression within replicating human B cells . Within inflammatory environments , B cells encountering foreign or self-Ag can develop tertiary lymphoid tissue expressing activation-induced cytosine deaminase ( Q9GZX7 ) . Recently , this DNA-modifying enzyme was detected in nonlymphoid cells within several inflamed tissues and strongly implicated in malignant transformation . This study examines whether a cyclooxygenase 2 ( P35354 ) pathway , often linked to inflammation , influences Q9GZX7 expression in activated B lymphocytes . In this paper , we report that dividing human B cells responding to surrogate C3d-coated Ag , P05112 , and Q9Y275 express Q9GZX7 , as well as P35354 . A progressive increase in Q9GZX7 with each division was paralleled by a division-related increase in a P35354 -linked enzyme , microsomal PGE(2) synthase-1 , and the PGE(2)R , EP2 . Cells with the greatest expression of Q9GZX7 expressed the highest levels of EP2 . Although P35354 inhibitors diminished both Q9GZX7 expression and IgG class switching , exogenous PGE(2) and butaprost , a selective EP2 agonist , augmented Q9GZX7 mRNA/protein and increased the numbers of IgG(+) progeny . Despite the latter , the proportion of IgG(+) cells within viable progeny generally declined with PGE(2) supplementation . This was not due to PGE(2)-promoted differentiation to plasma cells or to greater downstream switching . Rather , because phosphorylated ataxia telangiectasia mutated levels were increased in progeny of PGE(2)-supplemented cultures , it appears more likely that PGE(2) facilitates Q9GZX7 -dependent DNA double-strand breaks that block B cell cycle progression or promote activation-induced cell death , or both . Taken together , the results suggest that a PGE(2) feed-forward mechanism for augmenting P35354 pathway proteins promotes progressively increased levels of Q9GZX7 mRNA , protein , and function . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . Pan- Q07869 Agonist , DB01393 , Restores Angiogenesis in Hindlimb Ischemia in Normal and Diabetic Rats . Introduction. The aim of this study was to investigate the effect of bezafibrate as a pan- Q07869 agonist on angiogenesis and serum nitrite , the main metabolite of nitric oxide ( NO ) , vascular endothelial growth factor ( P15692 ) and P15692 receptor-2 ( P35968 ) concentrations in hindlimb ischemia model of normal and type I diabetic rats . Methods . 28 male Wistar rats were divided into control and diabetic groups . Then , all rats underwent unilateral hindlimb ischemia . After recovery , they were randomly assigned to one of the following experimental groups : ( 1 ) control ; ( 2 ) control + bezafibrate ( 400 mg/kg/day ) ; ( 3 ) diabetic ; ( 4 ) diabetic + beztafibrate . After three weeks , blood samples were taken and capillary density was evaluated in the gasterocnemius muscle of ischemic limb . Results . DB01393 increased capillary density and capillary/fiber ratio in ischemic leg of diabetic and control rats ( P < 0.05 ) . Serum P15692 and P35968 concentrations did not alter after bezafibrate administration , however , serum nitrite concentration was significantly higher in bezafibrate-treated groups than non-treated groups ( P < 0.05 ) . Discussion . It seems that bezafibrate , as a pan Q07869 agonist , restores angiogenesis in hindlimb ischemic diabetic animals and is useful for prevention and/or treatment of peripheral artery disease in diabetic subjects . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Detection of overexpressed P35354 in precancerous lesions of hamster pancreas and lungs by molecular imaging : implications for early diagnosis and prevention . The enzyme cyclooxygenase-2 ( P35354 ) is overexpressed in many cancers , cardiovascular disease , neurodegenerative disorders , and arthritis . Selective inhibitors of P35354 have been developed as therapeutics or preventive agents for these diseases . However , recent reports have revealed a significant increase in cardiovascular mortality in long-term users of the P35354 inhibitors Vioxx and DB00482 , emphasizing the need for noninvasive tests that allow the identification of individuals whose P35354 levels are overexpressed prior to assignment to treatment with these drugs . In this study , we have prepared a radioiodinated analogue of the selective P35354 inhibitor celecoxib , and verified its binding to the P35354 enzyme in vitro . Biodistribution studies in hamsters demonstrated significantly higher levels of radiotracer in animals treated with the tobacco carcinogen NNK in lung , pancreas , and liver . Assessment of P35354 levels by whole-body planar nuclear imaging two hours after injection of the radiotracer was suggestive of a distinct increase in P35354 in the pancreas and liver of a hamster treated for 10 weeks with NNK , in the lungs and liver of a second animal , and in the liver only , in two additional animals from the same treatment group . Immunostains showed selective overexpression of P35354 in pre-neoplastic lesions of the pancreas and lungs in only those animals that showed tracer accumulation in these organs and in the livers of all NNK-treated hamsters . Immunostains for P23219 yielded detectable reactions in the intestinal epithelium but not in pancreas , lungs , or liver , supporting the specificity of the tracer for P35354 . Our data provide proof of principle for the hypothesis that molecular imaging with radiolabeled P35354 inhibitors can be used for the noninvasive monitoring of overexpressed P35354 levels . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . P35354 inhibitors : do they have a role in emergency department prescribing ? P35354 selective inhibitors ( COXIB or CSI ) have been released with a fanfare as efficacious and safer alternatives to traditional non-steroidal anti-inflammatory drugs . They purport to offer equivalent degrees of analgesia and an improved safety profile . COXIB currently available in Australasia are celecoxib ( DB00482 ) , rofecoxib ( Vioxx ) and etoricoxib ( Arcoxia ) . This review discusses the pharmacology of these agents and reviews recent literature regarding their effectiveness and safety . It endeavours to answer the question ' Should we be using COXIB in emergency departments in Australasia ' ? Docking studies on NSAID/ P35354 isozyme complexes using contact statistics analysis . The selective inhibition of P35354 isozymes should lead to a new generation of NSAIDs with significantly reduced side effects ; e.g. celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . To obtain inhibitors with higher selectivity it has become essential to gain additional insight into the details of the interactions between P36551 isozymes-and NSAIDs . Although X-ray structures of P35354 complexed with a small number of ligands are available , experimental data are missing for two well-known selective P35354 inhibitors ( rofecoxib and nimesulide ) and docking results reported are controversial . We use a combination of a traditional docking procedure with a new computational tool ( Contact Statistics analysis ) that identifies the best orientation among a number of solutions to shed some light on this topic . Celecoxib-induced upper gastrointestinal hemorrhage and ulceration . P35354 inhibitors are a new class of nonsteroidal anti-inflammatory drugs with a reported benefit of less gastric and duodenal ulceration and hemorrhage . We describe a 67-year-old man taking a higher than usual dose of celecoxib ( DB00482 ) for osteoarthritis with resultant gastric erosions , ulceration , and a significant gastrointestinal ( GI ) hemorrhage . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . Enhanced killing of chemo-resistant breast cancer cells via controlled aggravation of ER stress . Moderate activity of the endoplasmic reticulum ( ER ) stress response system exerts anti-apoptotic function and supports tumor cell survival and chemoresistance , whereas its more severe aggravation may exceed the protective capacity of this system and turn on its pro-apoptotic module . In this study , we investigated whether the combination of two pharmacologic agents with known ability to trigger ER stress via different mechanisms would synergize and lead to enhanced tumor cell death . We combined the HIV protease inhibitor nelfinavir ( DB00220 ) and the cyclooxygenase 2 ( P35354 ) inhibitor celecoxib ( DB00482 ) and investigated their combined effect on ER stress and on the viability of breast cancer cells . We found that this drug combination aggravated ER stress and caused pronounced toxicity in human breast cancer cell lines , inclusive of variants that were highly resistant to other therapeutic treatments , such as doxorubicin , paclitaxel , or trastuzumab . The anti-tumor effects of celecoxib were mimicked at increased potency by its non-coxib analog , 2,5-dimethyl-celecoxib ( Q6UXB2 ) , but were substantially weaker in the case of unmethylated-celecoxib ( UMC ) , a derivative with superior P35354 inhibitory efficacy . We conclude that the anti-tumor effects of nelfinavir can be enhanced by celecoxib analogs in a P35354 independent fashion via the aggravation of ER stress , and such drug combinations should be considered as a beneficial adjunct to the treatment of drug-resistant breast cancers . Celecoxib and melatonin in prevention of female rat mammary carcinogenesis . The present experiment aims to evaluate tumor suppressive effects of a selective inhibitor of cyclooxygenase-2 ( P35354 ) celecoxib ( DB00482 , Pfizer ) administered alone and in combination with melatonin in the prevention of N-methyl-N-nitrosourea ( P48645 ) -induced mammary carcinogenesis in Sprague-Dawley female rats . Celecoxib was administered daily at a concentration of 1.666 g/kg diet to two groups during 20 weeks ( starting a week before first P48645 application ) . A combination of celecoxib and melatonin applied in drinking water ( 20 microg/ml drinking water ) , daily from 15:00 to 08:00 hours was administered to the second group . The anticarcinogenic effects of chemopreventive drugs were compared with control ( P48645 ) animals . Celecoxib administration decreased mammary tumor incidence ( by 24 % ) , while combination of celecoxib and melatonin decreased tumor incidence even more significantly ( -30 % ) . Significant decrease in tumor frequency per group was recorded in both groups with chemoprevention : celecoxib alone ( -54 % ) and combination of celecoxib and melatonin ( -64 % ) . Celecoxib significantly influenced tumor frequency per animal in the group with combination of both protective substances ( -52 % ) . Celecoxib administration resulted in prolonged latency by 3 % , and by 13 % in the group with combination of both protective substances . These results confirm preventive effects of celecoxib in induced rat mammary carcinogenesis . The administration of isolated P61006 had only lesser effect , but in the combination with CELE revealed some potentiating influence in mammary carcinogenesis inhibition . The present study is the first to prove efficacy of the above-mentioned celecoxib and melatonin intake . Our results point to the need for a deeper analysis of coxib efficacy in human carcinogenesis . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Gemcitabine/ DB00762 /celecoxib in pancreatic cancer . Unresectable pancreatic cancer has few therapeutic options and a dismal prognosis . P35354 ( P35354 ) expression is increased at the RNA and protein levels in most human pancreatic cancers . The purpose of this trial was to determine whether the addition of a P35354 inhibitor to chemotherapy was beneficial . To date , 11 patients with inoperable pancreatic cancer have been treated with the combination of gemcitabine ( Gemzar ) , irinotecan ( Camptosar ) , and celecoxib ( DB00482 ) at 400 mg orally twice daily . Encouraging pain relief , improvement in performance status , and decreases in CA 19-9 and carcinoembryonic antigen levels have been observed . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Microscopic modes and free energies of 3-phosphoinositide-dependent kinase-1 ( PDK1 ) binding with celecoxib and other inhibitors . Celecoxib , also known as DB00482 ( approved by FDA in 1998 ) and remembered as the fastest-selling drug in history , was used as a cyclooxygenase-2 ( P35354 ) selective inhibitor having both anti-inflammatory and anticancer activities . Most recent studies have revealed that the apoptotic activity of celecoxib ( and its derivatives ) is actually independent of the P35354 inhibitory activity and that celecoxib also inhibits the kinase activity of 3-phosphoinositide-dependent protein kinase-1 ( PDK1 ) , suggesting that the well-known anticancer activity of celecoxib is not due to the inhibition of P35354 , but possibly is due to the inhibition of PDK1 . It is highly desirable to develop new celecoxib derivatives as PDK1-specifc inhibitors to avoid the side effects of P35354 inhibitors . To understand how PDK1 binds with celecoxib and its derivatives , we have performed extensive molecular docking and combined molecular dynamics ( MD ) simulations and molecular mechanics/Poisson-Boltzmann surface area ( MM-PBSA ) binding free energy calculations on eight representative PDK1 inhibitors , leading to the finding of a new , more favorable binding mode which is remarkably different from the previously proposed binding mode . Based on the determined most stable binding structures , the calculated binding free energies are all in good agreement with the corresponding experimental data , and the biological activity data available for celecoxib and its derivatives can be better interpreted . The obtained new insights , concerning both the binding mode and computational protocol , will be valuable not only for future rational design of novel , more potent PDK1-specific inhibitors as promising anticancer therapeutics , but also for rational design of drugs targeting other proteins . P35354 inhibitors and DB00482 : safe or suspect ? Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . Effect of celecoxib on capecitabine-induced hand-foot syndrome and antitumor activity . We hypothesized that hand-foot syndrome is an inflammatory phenomenon mediated by the overexpression of cyclooxygenase 2 ( P35354 ) . Therefore , a specific P35354 inhibitor such as celecoxib ( DB00482 ) could attenuate both the incidence and severity of hand-foot syndrome . We undertook a retrospective study comparing the incidences of hand-foot syndrome in 67 patients with metastatic colorectal cancer who took capecitabine ( DB01101 ) with or without celecoxib . Surprisingly , celecoxib seemed to attenuate capecitabine-induced diarrhea as well . DB01101 /celecoxib was also associated with increased tumor response , proportion of stable disease ( 62.5 % vs 22.8 % , P = .001 ) , and increase in median time to tumor progression ( 6 vs 3 months , P = .002 ) compared with capecitabine alone , despite the fact that patients on capecitabine/celecoxib had less favorable disease characteristics ( age , performance status , and prior chemotherapies ) . Overexpression of P35354 , implicated in promoting angiogenesis , enhanced tumor invasiveness , evasion of apoptosis , and immune suppression , is a bona fide molecular target for many solid tumors , including colorectal cancer . Combining capecitabine with celecoxib in the treatment of colorectal cancer has strong preclinical rationales . A prospective study is being designed to evaluate capecitabine and celecoxib with or without epidermal growth factor receptor antagonist ZD1839 in the frontline treatment of metastatic colorectal cancer . These regimens under study are orally based and may significantly impact quality of life in the frontline treatment of metastatic colorectal cancer . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Dual carbonic anhydrase -- cyclooxygenase-2 inhibitors . Cyclooxygenase is a key enzyme responsible for metabolisation of arachidonic acid into prostaglandins and thromboxane . This enzyme is the target of non steroidal anti-inflammatory drugs ( NSAIDs ) , used against inflammation and pain . The inducible P35354 was associated with inflammatory conditions , whereas the constitutive form ( P23219 ) was responsible for the beneficial effects of the PGs . This observation led to the development of P35354 inhibitors or " coxibs " of which rofecoxib ( Vioxx ) characterized by a methylsulfone moiety and the sulfonamides celecoxib ( DB00482 ) and valdecoxib ( Bextra ) . Initially described as P35354 " selective " inhibitors , recent reports revealed a nanomolar inhibition activity of the sulfonamide P35354 inhibitors for several carbonic anhydrase ( CA ) isoforms , confirmed by X-ray crystal structures for the adducts of celecoxib and valdecoxib with isozyme CA II . This dual activity may help to explain differences in clinical observation between sulfonamide and methylsulfone P35354 inhibitors . Moreover , the inhibition of CA isozymes , critical for the development and invasion of cancer cells , such as CA II , IX and XII , may constitute an important mechanism of antitumor action of such sulfonamide compounds . P35354 inhibitors and cancer therapeutics : potential roles for inhibitors of P35354 in combination with cytotoxic therapy : reports from a symposium held in conjunction with the Radiation Therapy Oncology Group June 2001 Meeting . Tumor growth and angiogenesis are interdependent . Cyclooxygenase ( P36551 ) catalyzes the synthesis of prostaglandins ( PGs ) from arachidonic acid . Nonsteroidal antiinflammatory drugs ( NSAIDs ) inhibit P36551 -mediated synthesis of PGs . Although P23219 is constitutively expressed in a wide range of tissues , P35354 is cytokine inducible . Enhanced P35354 expression has been attributed a key role in the development of inflammation and related processes observed in pathologically altered disease states . Two specific P35354 inhibitors , namely , rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) , both oral agents and FDA approved , have been shown preclinically and clinically to have efficacy comparable to that of NSAIDs for relief of pain and inflammation in osteoarthritis , with decreased risk of gastrointestinal damage . Significant preclinical evidence strongly supports the potential role for these inhibitors for the treatment of cancer . On June 29 , 2001 , the Radiation Therapy Oncology Group ( www.rtog.org ) , a National Cancer Institute-sponsored cooperative group , held a 1-day symposium focusing on the potential role of inhibitors of P35354 in the treatment of cancer . This issue of the American Journal of Clinical Oncology contains the summary of those presentations . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE .

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Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . 17(E)-picolinylidene androstane derivatives as potential inhibitors of prostate cancer cell growth : antiproliferative activity and molecular docking studies . We report a rapid and efficient synthesis of A-ring modified 17α-picolyl and 17(E)-picolinylidene androstane derivatives from dehydroepiandrosterone . Compounds were validated spectroscopically and structurally characterized by X-ray crystallography . Virtual screening by molecular docking against clinical targets of steroidal anticancer drugs ( ERα , AR , P11511 and P05093 ) suggests that 17(E)-picolinylidene , but not 17α-picolyl androstanes could specifically interact with P05093 ( 17α-hydroxylase ) with similar geometry and affinity as DB05812 , a 17-pyridinyl androstane drug clinically used in the treatment of prostate cancer . In addition , several 17(E)-picolinylidene androstanes demonstrated selective antiproliferative activity against PC3 prostate cancer cells , which correlates with DB05812 antiproliferative activity and predicted P05093 binding affinities . Based on these preliminary results , 17(E)-picolinylidene androstane derivatives could be a promising starting point for the development of new compounds for the treatment of prostate cancer . Particle size of latex beads dictates IL-1β production mechanism . Macrophages ( Mϕ ) are well documented to produce IL-1β through various signaling pathways in response to small particles such as silica , asbestos and urea crystals , in the presence of lipopolysaccharide ( LPS ) . However , it has not been clear to what extent particle size affects the response . To investigate this point , we stimulated bone marrow-derived macrophages ( BMDM ) with size-defined latex beads ( LxB ) . Although both nano-sized ( 20 nm ) and micro-sized ( 1,000 nm ) LxB induced IL-1β production , only the nano-sized particles formed large intracellular vacuoles . In contrast , 100 nm LxB did not induce either of the responses . The same cellular responses were also observed in primary microglia cells . Although K(+) efflux and Q96P20 activation in BMDM were crucial in response to both 20 and 1,000 nm LxB , only IL-1β production by 20 nm LxB was sensitive to cathepsin B and Q99572 , a receptor for DB00171 . The response by 1,000 nm LxB relied on a robust production of reactive oxygen species ( ROS ) , since IL-1β production was remarkably reduced by ROS inhibitors such as diphenylene iodonium ( DPI ) and DB06151 ( Q9C000 ) . In contrast , IL-1β production by 20 nm LxB was augmented by Q9C000 and in BMDM deficient in thioredoxin-binding protein-2 ( P20226 -2 ) , a negative regulator of the ROS scavenger thioredoxin . These results suggest that the cells responded differently in their secretion of IL-1β depending on particle size , and that there is a range within which neither pathway works . P10275 levels are upregulated by Akt in prostate cancer . Multiple lines of evidence suggest a functional link between the androgen receptor ( AR ) and the serine/threonine kinase Akt in the development and progression of prostate cancer . To investigate the impact of Akt activity on AR homeostasis , we treated androgen-dependent LNCaP and LAPC-4 prostate cancer cells with Akt inhibitor . Akt inhibition decreased AR expression , suggesting that Akt activity was required for regulation of AR protein levels . However , while androgen-independent LNCaP-abl cells also showed diminished AR protein levels in response to Akt inhibition , treatment of androgen-independent LNCaP-AI cells failed to alter AR protein levels upon similar treatment , suggesting that AR protein levels in these androgen-independent prostate cells were regulated by mechanisms independent of Akt activation . Regulation of AR , downstream of activated Akt , also was observed in vivo when examining transgenic mice that overexpress constitutively active mutant myristoylated (myr)-Akt1 in the prostate . Transgenic mice expressing activated myr-Akt1 exhibited higher levels of AR mRNA and protein . Expression of activated myr-Akt1 did not alter prostate cell growth and no significant size differences between prostate tissues derived from transgenic animals were observed when comparing transgenic mice with wild-type mice . Still , transgenic mice overexpressing Akt exhibited higher levels of γ P16104 and phosphorylated Chk2 in prostate tissue . These changes in markers associated with oncogene-induced senescence confirmed significant altered signaling in the transgenic mouse model . Overall , results presented here suggest that AR levels are regulated by the Akt pathway . Androgen synthesis inhibitors in the treatment of castration-resistant prostate cancer . Suppression of gonadal testosterone synthesis represents the standard first line therapy for treatment of metastatic prostate cancer . However , in the majority of patients who develop castration-resistant prostate cancer ( CRPC ) , it is possible to detect persistent activation of the androgen receptor ( AR ) through androgens produced in the adrenal gland or within the tumor itself . DB05812 acetate was developed as an irreversible inhibitor of the dual functional cytochrome P450 enzyme P05093 with activity as a 17α-hydroxylase and 17,20-lyase . P05093 is necessary for production of nongonadal androgens from cholesterol . Regulatory approval of abiraterone in 2011 , based on a phase III trial showing a significant improvement in overall survival ( OS ) with abiraterone and prednisone versus prednisone , represented proof of principle that targeting AR is essential for improving outcomes in men with CRPC . Inhibition of 17α-hydroxylase by abiraterone results in accumulation of upstream mineralocorticoids due to loss of cortisol-mediated suppression of pituitary adrenocorticotropic hormone ( DB01285 ) , providing a rationale for development of P05093 inhibitors with increased specificity for 17,20-lyase ( orteronel , galeterone and VT-464 ) that can potentially be administered without exogenous corticosteroids . In this article , we review the development of abiraterone and other P05093 inhibitors ; recent studies with abiraterone that inform our understanding of clinical parameters such as drug effects on quality-of-life , potential early predictors of response , and optimal sequencing of abiraterone with respect to other agents ; and results of translational studies providing insights into resistance mechanisms to P05093 inhibitors leading to clinical trials with drug combinations designed to prolong abiraterone benefit or restore abiraterone activity . P05093 inhibitors in castration-resistant prostate cancer . The majority of prostate cancer ( PCa ) cases are diagnosed as a localized disease . Definitive treatment , active surveillance or watchful waiting are employed as therapeutic paradigms . The current standard of care for the treatment of metastatic PCa is either medical or surgical castration . Once PCa progresses in spite of castrate androgen levels it is termed ' castration-resistant prostate cancer ' ( CRPC ) . Patients may even exhibit rising PSA levels with possible bone , lymph node or solid organ metastases . In 2010 , the only agent approved for the treatment of CRPC was docetaxel , a chemotherapeutic agent . It is now known that cells from patients with CRPC express androgen receptors ( AR ) and remain continuously influenced by androgens . As such , treatments with novel hormonal agents that specifically target the biochemical conversion of cholesterol to testosterone have come to the forefront . The use of cytochrome P450c17 ( P05093 ) inhibitor underlies one of the most recent advances in the treatment of CRPC . DB05812 acetate ( AA ) was the first P05093 inhibitor approved in the United States . This review will discuss CRPC in general with a specific focus on AA and novel P05093 inhibitors . AA clinical trials will be reviewed along with other novel adjunct treatments that may enhance the effectiveness of abiraterone therapy . Furthermore , the most recently identified P05093 inhibitors Orteronel , Galeterone , VT-464 , and CFG920 will also be explored . Novel P05093 inhibitors : synthesis , biological evaluation , structure-activity relationships and modelling of methoxy- and hydroxy-substituted methyleneimidazolyl biphenyls . Recently , the steroidal P05093 inhibitor DB05812 entered phase II clinical trial for the treatment of androgen-dependent prostate cancer . As 17alpha-hydroxylase-17,20-lyase ( P05093 ) catalyzes the last step in androgen biosynthesis , inhibition of this target should affect not only testicular but also adrenal androgen formation . Therefore P05093 inhibitors should be advantageous over existing therapies , for example with DB00644 analogues . However , steroidal drugs are known for side effects which are due to affinities for steroid receptors . Therefore we decided to synthesize non-steroidal compounds mimicking the natural P05093 substrates pregnenolone and progesterone . The synthesis and biological evaluation of a series of 15 novel and highly active non-steroidal P05093 inhibitors are reported . The compounds were prepared via Suzuki-cross-coupling , Grignard reaction and CDI-assisted S(N)t-reaction with imidazole and their inhibitory activity was examined with recombinant human P05093 expressed in Escherichia coli . Promising compounds were further tested for their selectivity against the hepatic enzyme P08684 and the glucocorticoid-forming enzyme P15538 . All compounds turned out to be potent P05093 inhibitors . The most active compounds 7 and 8 were much more active than Ketoconazole showing activity comparable to DB05812 ( IC(50) values of 90 and 52nM vs. 72nM ) . Most compounds also showed higher selectivities than Ketoconazole , but turned out to be less selective than DB05812 . Docking studies using our P05093 protein model were performed with selected compounds to study the interactions between the inhibitors and the amino acid residues of the active site . Mammalian Q99572 receptor pharmacology : comparison of recombinant mouse , rat and human Q99572 receptors . BACKGROUND AND PURPOSE : Acute activation of Q99572 receptors rapidly opens a non-selective cation channel . Sustained Q99572 receptor activation leads to the formation of cytolytic pores , mediated by downstream recruitment of hemichannels to the cell surface . Species- and single-nucleotide polymorphism-mediated differences in Q99572 receptor activation have been reported that complicate understanding of the physiological role of Q99572 receptors . Studies were conducted to determine pharmacological differences between human , rat and mouse Q99572 receptors . EXPERIMENTAL APPROACH : Receptor-mediated changes in calcium influx and Yo-Pro uptake were compared between recombinant mouse , rat and human Q99572 receptors . For mouse Q99572 receptors , wild-type ( BALB/c ) and a reported loss of function ( C57BL/6 ) Q99572 receptor were also compared . KEY RESULTS : BzATP [ 2,3-O-(4-benzoylbenzoyl)- DB00171 ] was more potent than DB00171 in stimulating calcium influx and Yo-Pro uptake at rat , human , BALB/c and C57BL/6 mouse Q99572 receptors . Two selective Q99572 receptor antagonists , A-740003 and A-438079 , potently blocked Q99572 receptor activation across mammalian species . Several reported P51575 receptor antagonists [ e.g. P59665 2159 ( 4- [ ( 4-formyl-5-hydroxy-6-methyl-3- [ (phosphonooxy)methyl } -2-pyridinyl ) azo ] -benzoic acid ) , PPNDS and NF279 ] blocked Q99572 receptors . NF279 fully blocked human Q99572 receptors , but only partially blocked BALB/c Q99572 receptors and was inactive at C57BL/6 Q99572 receptors . CONCLUSIONS AND IMPLICATIONS : These data provide new insights into Q99572 receptor antagonist pharmacology across mammalian species . Q99572 receptor pharmacology in a widely used knockout background mouse strain ( C57BL/6 ) was similar to wild-type mouse Q99572 receptors . Several structurally novel , selective and competitive Q99572 receptor antagonists show less species differences compared with earlier non-selective antagonists . DB05812 acetate : in metastatic castration-resistant prostate cancer . Oral abiraterone acetate , in combination with prednisone/prednisolone , is used to treat patients with metastatic castration-resistant prostate cancer ( CRPC ) who have previously received docetaxel-containing chemotherapy . DB05812 acetate was developed to specifically inhibit cytochrome P450 (CYP)17A1 , which is an essential enzyme in the biosynthesis of testosterone . In a pivotal phase III trial in patients with metastatic CRPC who have previously received docetaxel-containing chemotherapy , abiraterone acetate 1000 mg once daily plus prednisone 5 mg twice daily significantly prolonged overall survival compared with placebo plus prednisone . In this trial , abiraterone acetate plus prednisone was significantly more effective than placebo plus prednisone in prolonging the time to prostate-specific antigen ( PSA ) progression and in prolonging progression-free survival . Significantly more abiraterone acetate plus prednisone recipients than placebo plus prednisone recipients were considered to be responders , when assessed by PSA levels or radiographic imaging . Treatment with abiraterone acetate plus prednisone in the phase III trial was associated with an acceptable tolerability profile , which was generally similar to that of the placebo plus prednisone group . However , adverse events of special interest ( e.g. cardiac disorders and liver-function test abnormalities and adverse events resulting from elevated mineralocorticoid levels because of P05093 inhibition [ i.e. fluid retention and oedema , hypokalaemia , hypertension ] ) occurred in significantly more abiraterone acetate plus prednisone than in placebo plus prednisone recipients . HIV endocytosis after dendritic cell to T cell viral transfer leads to productive virus infection . Contacts between HIV-producing T cells and primary P01730 + T cells may induce the uptake of HIV by target cells that are endocytosed into trypsin-resistant compartments . We have now compared the mechanism of virus transmission from T cell-to-T cell versus infected dendritic cells ( DCs ) -to-T cell . In cocultures of HIV-1-infected DCs with primary P01730 + T cells , virus transmission to the target cells was resistant to trypsin treatment and could only be prevented by the anti-SUgp120 antibody IgGb12 but not by P50750 -779 , C34 or DB00495 . Importantly , upon stimulation of purified HIV-1-loaded P01730 + T cells with PHA/ P60568 , cells became productively infected as measured by intracellular CAp24 staining and antigen determination in the cell supernatant . These results suggest that the viral endocytic transfer may represent a escape mechanism in the presence of drugs targeting HIV-1 entry or the host immune system . Interactions of abiraterone , eplerenone , and prednisolone with wild-type and mutant androgen receptor : a rationale for increasing abiraterone exposure or combining with MDV3100 . Prostate cancer progression can be associated with androgen receptor ( AR ) mutations acquired following treatment with castration and/or an antiandrogen . DB05812 , a rationally designed inhibitor of P05093 recently approved for the treatment of docetaxel-treated castration-resistant prostate cancer ( CRPC ) , is often effective , but requires coadministration with glucocorticoids to curtail side effects . Here , we hypothesized that progressive disease on abiraterone may occur secondary to glucocorticoid-induced activation of mutated AR . We found that prednisolone plasma levels in patients with CRPC were sufficiently high to activate mutant AR . P08235 antagonists , such as spironolactone and eplerenone that are used to treat side effects related to mineralocorticoid excess , can also bind to and activate signaling through wild-type or mutant AR . DB05812 inhibited in vitro proliferation and AR-regulated gene expression of AR-positive prostate cancer cells , which could be explained by AR antagonism in addition to inhibition of steroidogenesis . In fact , activation of mutant AR by eplerenone was inhibited by MDV3100 , bicalutamide , or greater concentrations of abiraterone . Therefore , an increase in abiraterone exposure could reverse resistance secondary to activation of AR by residual ligands or coadministered drugs . Together , our findings provide a strong rationale for clinical evaluation of combined P05093 inhibition and AR antagonism . P05093 inhibition as a hormonal strategy for prostate cancer . P10275 ( AR ) signaling has a key role in the pathogenesis of prostate cancer . AR gene amplification , AR overexpression , and activating mutations in the AR occur more frequently as castration-resistant prostate cancer ( CRPC ) evolves , with intratumoral androgen levels remaining sufficient for AR activation despite castration . The source of these androgens might be either adrenal or intratumoral . AR signaling , therefore , remains a valid treatment target for patients with CRPC . P05093 is a key enzyme for androgen biosynthesis . The imidazole antifungal agent ketoconazole weakly and nonspecifically inhibits P05093 , but remains unlicensed for this indication . Chemists at the Cancer Research UK Centre for Cancer Therapeutics have designed a novel , selective , irreversible inhibitor of P05093 called abiraterone , which is more than 20 times more potent than ketoconazole . DB05812 acetate , a prodrug , has undergone phase I assessment , and is rapidly progressing from phase II to phase III trials , in view of its high level of antitumor activity . This agent is safe and well tolerated , and activity profiles suggest that approximately 50 % of CRPC remains AR-ligand driven . Other P05093 inhibitors with alternative mechanisms of action , for example VN/124-1 , are in preclinical development . The rationale for and implications of P05093 inhibition and the P05093 -targeting agents in development are discussed in this Review . Chronic exposure to environmental levels of tribromophenol impairs zebrafish reproduction . Tribromophenol ( 2,4,6- P20226 ) is ubiquitously found in aquatic environments and biota . In this study , we exposed zebrafish embryos ( F(0) ; 2 " " days post-fertilization , dpf ) to environmental concentration ( 0.3 microg/L ) and a higher concentration ( 3.0 microg/L ) of P20226 and assessed the impact of chronic exposure ( 120 dpf ) on reproduction . P20226 exposure did not cause a significant increase in the malformation and reduction in the survival in the F(0)-generation fish . After P20226 exposure , the plasma testosterone and estradiol levels significantly increased in males and decreased in females . The transcription of steroidogenic genes ( 3beta-HSD , 17beta-HSD , P05093 , CYP19A , CYP19B ) was significantly upregulated in the brain and testes in males and downregulated in the brain and ovary in females . P20226 exposure significantly downregulated and upregulated the expression of VTG in the liver of female and male fish , respectively . Meanwhile , P20226 exposure altered the sex ratio toward a male-dominant state . The F(1)-generation larvae exhibited increased malformation , reduced survival , and retarded growth , suggesting that P20226 in the aquatic environment has significant adverse effects on fish population . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . The 5α-androstanedione pathway to dihydrotestosterone in castration-resistant prostate cancer . The survival and progression of prostate cancer are generally dependent on expression of the androgen receptor ( AR ) , as well as the availability of endogenous AR agonists . Originating from the gonads , testosterone is released into circulation and is converted by steroid-5α-reductase in prostate cancer to 5α-dihydrotestosterone ( DB02901 ) , potently activating AR and driving tumor progression . Advanced prostate cancer is initially treated with gonadal testosterone depletion , which suppresses this cascade of events and typically leads to a treatment response . Eventually , resistance to testosterone deprivation occurs with " castration-resistant " prostate cancer ( CRPC ) and is driven by the intratumoral synthesis of DB02901 . The generation of DB02901 occurs in large part from adrenal 19-carbon precursor steroids , which are dependent on expression of P05093 . Although the path from adrenal precursor steroids to DB02901 was generally thought to require 5α-reduction of testosterone , recent data suggest that it instead involves conversion from Δ-androstenedione by steroid-5α-reductase isoenzyme-1 to 5α-androstanedione , followed by subsequent conversion to DB02901 . The 5α-androstanedione pathway to DB02901 therefore bypasses testosterone entirely . DB05812 acetate effectively inhibits P05093 , blocks the synthesis of androgens , and extends the survival of men with CRPC . Further progress in the hormonal treatment of CRPC is dependent on an understanding of the mechanisms that underlie CRPC and resistance to abiraterone acetate . P19957 kinase p110β : a therapeutic target in advanced prostate cancers . Prostate cancers in the castration-resistant stage are life-threatening because they are not curable in clinic . The novel androgen receptor inhibitor Xandi ( DB08899 ) and the new P05093 inhibitor Zytiga ( DB05812 ) prolonged patient survival only a few months in advanced prostate cancers . Therefore , novel therapeutic agents for advanced prostate cancers are urgently needed . P19957 kinases are major intracellular signaling molecules that regulate multiple signal pathways related to cellular metabolism , cytokinesis , growth and survival . Accumulating evidence in the literature indicates that some isoforms of this kinase family are oncogenic and abnormally expressed in various human cancers , including prostate cancers . Recent extensive studies from our group and others showed that P19957 kinase p110β is aberrantly overexpressed in advanced prostate cancers and is critical for prostate cancer development and progression as demonstrated in cell-based and animal models . Importantly , novel p110β-specific inhibitors have been developed and are currently been testing in clinical trials . In this article , we will briefly summarize recent developments in this regard . Is abiraterone acetate well tolerated and effective in the treatment of castration-resistant prostate cancer ? This Practice Point commentary discusses the findings of the first phase I trial to evaluate abiraterone acetate ( an inhibitor of the androgen-regulating enzyme P05093 ) in the treatment of castration-resistant prostate cancer . This open-label , dose-escalation study by Attard et al. showed that abiraterone was well tolerated but often induced a syndrome of secondary mineralocorticoid excess that improved with eplerenone ( a mineralocorticoid receptor antagonist ) . DB05812 is a potent suppressor of adrenal androgen synthesis , and produced lasting prostate-specific antigen responses in approximately half of the patients . A few patients had partial regression of distant metastases . Although promising , these results should be interpreted with caution owing to the small sample size and because the study was not primarily designed to examine drug efficacy . Multi-institutional , prospective trials should provide additional information on the tolerability and activity of this compound and further define the population most likely to benefit from this endocrine approach . DB05812 in heavily pretreated patients with metastatic castrate-resistant prostate cancer . The aim of this study was to evaluate the activity and tolerability of abiraterone acetate in patients with metastatic castrate-resistant prostate cancer treated previously with more than three lines of chemotherapy . Patients received 1 g of abiraterone acetate ( administered as four 250 mg tablets ) orally once daily with prednisone at a dose of 5 mg orally twice daily . The primary endpoint was prostate-specific antigen ( PSA ) response . From August 2011 to January 2013 , 36 patients were enrolled . PSA response was observed in 22 patients ( 61.1 % , 95 % confidence interval : 0.41-0.81 ) . The median time to PSA progression was 7.3 months and after a median follow-up of 10.1 months , all patients were alive . The treatment was generally well tolerated ; side effects secondary to mineralocorticoid excess resulting from blockade of P05093 were largely controlled with prednisone . DB05812 acetate seems to be an effective and well-tolerated treatment option for patients with metastatic castrate-resistant prostate cancer irrespective of the number of chemotherapy lines administered previously . Association of the polymorphisms of genes involved in androgen metabolism and signaling pathways with familial prostate cancer risk in a Japanese population . BACKGROUND : Androgen plays a central role in the normal and malignant development of prostate glands . Genetic polymorphisms of genes involved in androgen metabolism and signaling might be associated with the risk of prostate cancer . METHODS : One hundred and two patients with prostate cancer with a family history and 117 healthy age- and residence-matched male controls were enrolled . Genotypes of the CAG repeat length of androgen receptor ( AR ) , P05093 , 5alpha-reductase type II ( P31213 ) , P13051 -glucuronosyltransferase ( P78381 ) 2B15 , PSA promoter genes were analyzed . RESULTS : For single polymorphisms , the presence of Y alleles showed a significantly lower risk of prostate cancer in comparison with the D/D genotype in P54855 ( odds ratio [OR]=0.41 , 95 % confidence interval [CI]=1.40-4.28 , p=0.0015 ) , and the presence of A2 alleles showed a weak tendency to decrease prostate cancer risk in comparison with the A1/A1 genotype in P05093 ( OR=0.69 , 95 % CI=0.39-1.23 , p=0.21 ) . The stratification of cases according to clinical stage and pathological grade showed that the A2/A2 genotype was significantly associated with localized stage cancer in comparison with metastatic stage cancer ( OR=5.18 , 95 % CI=1.49-17.95 , p=0.007 ) . The combination of P54855 and P05093 genotypes could identify higher risk subjects even in subjects with low-risk P54855 genotypes , i.e. , Y/Y+D/Y genotypes ( OR=1.97 , 95 % CI=0.92-4.22 , p=0.079 ) . CONCLUSION : Genetic polymorphisms of the genes involved in androgen metabolism and signaling were significantly associated with familial prostate cancer risk . Single nucleotide polymorphisms of low-penetrance genes could be targets to understand genetic susceptibility to familial prostate cancer . Limited in vitro efficacy of P05093 inhibition on human castration resistant prostate cancer . Although accumulating evidence indicates high expression of P05093 (P45017A1) allows castration resistant prostate cancer ( CRPC ) to maintain high intratumoral androgen levels , the potential P45017A1 activity has not been characterized yet . The aim of this study was to examine the potential P05093 activity including 17α-hydroxylase and 17,20-lyase activities in human CRPC and the effect of a CYP17A inhibitor . We used three human CRPC cell lines : C4-2 and C4-2AT6 which was established from C4-2 under androgen ablation conditions for 6months , and PC3 . To ascertain the potential P05093 activity , we cultured with the steroid precursors : (13)C-[2,3,4]-progesterone ( 13C-Prog ) , and analyzed the sequential biosynthesis (13)C-[2,3,4]-17-hydroxyprogesterone ( 13C-17OHP ) and (13)C-[2,3,4]-androstenedione(13C-Adione) by liquid chromatography/mass spectrometry (LC/MS/MS).The C4-2AT6 cells showed significantly higher P05093 expression than C4-2 cells ( p < 0.001 ) . LC/MS/MS analysis enabled us to detect the 13C-17-OHP and 13C-A-dione in these cell lines . The concentration ratio of 13C-Adione/13C-17OHP ( Adione-17OHP ratio ) , which is thought to reflect the differences between 17-hydroxylase and 17,20-lyase activities , was then determined . The Adione-17OHP ratio in C4-2AT6 cells was significantly higher than that of C4-2 cells ( p < 0.001 ) . DB05812 were able to inhibit the CYP17A activities , although abiraterone did not have anti-proliferative effects on C4-2 and C4-2AT6 cells at clinically achievable concentrations of < 1000nM in vitro . The present study clearly demonstrates CRPC have the dual activities of P05093 mediated by 17-hydroxylase activity and 17,20-lyase activity . DB05812 does n't have an in vitro anti-proliferative efficacy in CRPC cells , suggesting limited efficacy in vitro . Effect of abiraterone acetate plus prednisone on the QT interval in patients with metastatic castration-resistant prostate cancer . PURPOSE : DB05812 is the active metabolite of the pro-drug abiraterone acetate ( AA ) and a selective inhibitor of P05093 , a key enzyme in testosterone synthesis , and improves overall survival in postdocetaxel metastatic castration-resistant prostate cancer ( mCRPC ) . This open-label , single-arm phase 1b study was conducted to assess the effect of AA and abiraterone on the QT interval . METHODS : The study was conducted in 33 patients with mCRPC . Patients received AA 1,000 mg orally once daily + prednisone 5 mg orally twice daily . Electrocardiograms ( ECGs ) were collected in triplicate using 12-lead Holter monitoring . Baseline ECGs were obtained on Cycle 1 Day-1 . Serial ECG recordings and time-matched pharmacokinetic ( PK ) blood samples were collected over 24 h on Cycle 1 Day 1 and Cycle 2 Day 1 . Serial PK blood samples were also collected over 24 h on Cycle 1 Day 8 . RESULTS : After AA administration , the upper bound of the 2-sided 90 % confidence interval ( CI ) for the mean baseline-adjusted QTcF change was < 10 ms ; no patients discontinued due to QTc prolongation or adverse events . No apparent relationship between change in QTcF and abiraterone plasma concentrations was observed [ estimated slope ( 90 % CI ) : 0.0031 ( -0.0040 , 0.0102 ) ] . CONCLUSIONS : There is no significant effect of AA plus prednisone on the QT/QTc interval in patients with mCRPC . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . Synthesis , biological evaluation , and molecular modeling studies of methylene imidazole substituted biaryls as inhibitors of human 17alpha-hydroxylase-17,20-lyase ( P05093 ) -- part II : Core rigidification and influence of substituents at the methylene bridge . Thirty-five novel substituted imidazolyl methylene biphenyls have been synthesized as P05093 inhibitors for the potential treatment of prostate cancer . Their activities have been tested with recombinant human P05093 expressed in Escherichia coli . Promising compounds were tested for selectivity against P15538 , P19099 , and hepatic CYP enzymes 3A4 , 1A2 , 2B6 and 2D6 . The core rigidified compounds ( 30-35 ) were the most active ones , being much more potent than Ketoconazole and reaching the activity of DB05812 . However , they were not very selective . Another rather potent and more selective inhibitor ( compound 23 , IC(50)=345 nM ) was further examined in rats regarding plasma testosterone levels and pharmacokinetic properties . Compared to the reference DB05812 , 23 was more active in vivo , showed a longer plasma half-life ( 10h ) and a higher bioavailability . Using our P05093 homology protein model , docking studies with selected compounds were performed to study possible interactions between inhibitors and amino acid residues of the active site . [ DB01373 signaling mediated by nicotine receptors in neurons ] . DB00184 has many acute and chronic pharmacological effects . DB00184 treatment activates neuronal nicotinic acetylcholine receptors ( nAChR ) in peripheral and central nervous systems leading to depolarization and elevation of intracellular calcium levels , which are considered to cause stimulation of neurotransmitter release , synaptic transmission , intracellular signal transduction and gene expression . Multiple subtypes of nAChRs display different sensitivity to nicotinic agonists and antagonists . Each of these subtypes has a unique distribution in peripheral and central nervous systems . Although presynaptic nAChRs have been extensively studied to modulate the release of neurotransmitters , the functional importance of nAChRs in somata is not sufficiently characterized . To clarify the mechanisms of calcium signaling and its stimulation of gene expression via nAChRs in somata , we have investigated nAChR-mediating calcium signaling mechanisms including phosphorylation of Q8NFH3 /44 Q96HU1 kinase ( P29323 ) , CREB and Akt in PC12h cells . DB00184 transiently activates phosphorylation of P29323 - , CREB and Akt . DB00184 induces the activation of both P19957 kinase/Act and P29323 /CREB pathways via common pathways including non-alpha 7-nAChRs , L-type VSCC , P62158 kinase and P00533 in PC12h cells , but Src family tyrosine kinases only participate in the pathway to activate Akt . Based on these results , we discuss nAChR signaling mechanisms in neurons . Interleukins 1α and 1β as regulators of steroidogenesis in human NCI-H295R adrenocortical cells . Inflammatory cytokines interleukin-1 ( IL-1 ) and tumor necrosis factor-α ( P01375 -α ) regulate the activity of the hypothalamo-pituitary-adrenal ( Q9Y251 ) axis at several levels . Although hypothalamic P06850 secretion may be the primary mechanism by which these cytokines activate the Q9Y251 axis , IL-1 expression is increased within the adrenal glands in models for systemic inflammation , and IL-1 may augment adrenal glucocorticoid production . Our aim was to investigate the direct effects of IL-1α and IL-1β on adrenal steroidogenesis and expression of three key steroidogenic genes in human adrenocortical cells using the NCI-H295R cell line as a model . mRNAs encoding receptors for IL-1 , P01375 -α , and leukemia inhibitory factor ( P15018 ) were detectable in the cell line ( Affymetrix microarray analysis ) . Both IL-1α and IL-1β increased cortisol , androstenedione , dehydroepiandrosterone and DB05804 production , and the accumulation of mRNAs for steroidogenic acute regulatory protein ( STAR ) , 17α-hydroxylase/17,20-lyase ( P05093 ) and 3β-hydroxysteroid dehydrogenase 2 ( P26439 ) in these cells ( P < 0.05 for all ) . Both ILs augmented P01375 -α- and P15018 -induced STAR and P05093 mRNA accumulation , and P01375 -α-induced cortisol production ( P < 0.05 for all ) . Both ILs also increased the apoptotic index of the cells ( P < 0.05 ) , which was efficiently neutralized by their specific antibodies . The IL-induced changes in the STAR , P26439 , and P05093 protein levels were not as evident as those in the respective mRNA levels . In conclusion , the combined effect of inflammatory cytokines at the adrenal level in acute or chronic inflammatory states could significantly stimulate glucocorticoid production , and thus explain the observed discrepancy between the cortisol and DB01285 concentrations sometimes seen in sepsis and chronic inflammatory states . Beyond castration and chemotherapy : novel approaches to targeting androgen-driven pathways . In castrate-resistant prostate cancer , beyond chemotherapy , existing guidelines suggest only supportive care . However , recent evidence suggests that continued targeting of androgen-dependent pathways may be an efficacious approach . Clinical data is now available for two mechanistically distinct agents ( abiraterone and MDV3100 ) that both ultimately target these pathways . DB05812 is a potent and irreversible inhibitor of P05093 , a critical enzyme in androgen biosynthesis . Phase II studies indicate substantial declines in PSA amongst castrate-resistant patients treated with abiraterone , both prior to and following cytotoxic chemotherapy . In contrast to abiraterone , MDV3100 is a direct inhibitor of the androgen receptor , binding the receptor irreversibly with substantially higher affinity as compared to bicalutamide . A recent phase I/II trial of MDV3100 in castrate-resistant prostate cancer demonstrated tolerability of the agent with activity at the lowest dose level . On the basis of these compelling data , both abiraterone and MDV3100 will be examined in the phase III setting . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . P38398 -associated breast and ovarian cancer risks in Poland : no association with commonly studied polymorphisms . Polymorphisms in genes involved in DNA repair , steroid hormone biosynthesis/metabolism/signaling , folate metabolism as well as cell growth are prime candidates for possible associations with breast and ovarian cancer risk in women with an inherited predisposition . We investigated 29 polymorphisms in 20 genes encoding key proteins of the above four biological pathways for their breast and ovarian cancer risk modifying effect in Polish women harboring P38398 founder mutations . Of the analyzed genes , P18074 , P18887 , O43543 , O43542 and Lig4 participate in DNA repair , P04637 in cell cycle check point control , Q9Y6Q9 , AR , P21964 , P05108 , P05093 , P11511 , Q8NA42 and P06401 in steroid hormone biosynthesis/metabolism/signaling , P04818 in folate metabolism and P04626 , P05231 , Q07954 , P01137 and P36897 affect cell growth . Using validated methods , we genotyped 319 breast cancer cases , 146 ovarian cancer cases and 290 unaffected controls , all of whom harbored one of three causative mutations in P38398 . Our results revealed no association of any of the investigated polymorphisms with P38398 -associated breast or ovarian cancer risk . Thus , it appears that these polymorphisms do not influence disease risk in Polish women carrying one of the three common P38398 founder mutations . DB05812 inhibits 1α,25-dihydroxyvitamin D3 metabolism by P08684 in human liver and intestine in vitro . The chemopreventive and therapeutic effects of vitamin D3 are exerted through its dihydroxylated metabolite , 1α,25-dihydroxyvitamin D3 [ 1α,25(OH)2D3 ] . Inactivation of 1α,25(OH)2D3 by cytochrome P450 3A4 ( P08684 ) may be an important determinant of its serum and tissue levels . DB05812 , a steroidogenesis inhibitor used in late stage prostate cancer treatment , is a P05093 inhibitor . The purpose of this study was to assess the potential of abiraterone to block hepatic and intestinal inactivation of biologically active vitamin D3in vitro and to evaluate if abiraterone can alter P08684 marker substrate activities . Biotransformation reactions were initiated with NADPH regenerating solutions following initial preincubation of pooled human hepatic or intestinal microsomal protein or human recombinant P08684 supersomes with 1α,25(OH)2D3 , midazolam or triazolam for 10min at 37°C . Formation of hydroxylated metabolites of 1α,25(OH)2D3 , midazolam or triazolam was analyzed by liquid chromatography-mass spectrometry method . Co-incubation of 1α,25(OH)2D3 with abiraterone at varying concentrations ( 0.2-100μM ) led to up to ∼85 % inhibition of formation of hydroxylated metabolites of 1α,25(OH)2D3 thus preventing inactivation of active vitamin D3 . The IC50 values for individual metabolites of 1α,25(OH)2D3 ranged from 0.4 to 2.2μM in human liver microsomes or human intestinal microsomes . The mechanism of P08684 -mediated inhibition of 1α,25(OH)2D3 by abiraterone was competitive ( apparent Ki 2.8-4.3μM ) . Similar inhibitory effects were also observed upon inclusion of abiraterone into midazolam or triazolam hydroxylation assays . In summary , our results suggest that abiraterone inhibits the P08684 -mediated inactivation of active vitamin D3 in human liver and intestine , potentially providing additional anti-cancer benefits to prostate cancer patients . This article is part of a Special Issue entitled ' 16th Vitamin D Workshop ' . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . DB05812 acetate : a promising drug for the treatment of castration-resistant prostate cancer . DB05812 acetate ( CB7630 ) , a pregnenolone analog , is an orally administered small molecule that irreversibly inhibits a rate-limiting enzyme in androgen biosynthesis , P05093 , and blocks the synthesis of androgens in the testes , adrenal glands and prostate without causing adrenal insufficiency . In clinical studies , abiraterone acetate is well tolerated and shows promising clinical activity in castration-resistant prostate cancer . The recommended Phase II dose of abiraterone acetate is 1000 mg orally daily in combination with prednisone 5 mg twice daily . Side effects are minimal and mostly associated with secondary mineralocorticoid excess , owing to a compensatory increase in upstream steroids , such as deoxycorticosterone and corticosterone . These include hypertension , hypokalemia and edema and are easily manageable with a selective mineralocorticoid antagonist , such as eplerenone , or low-dose corticosteroids . Currently , abiraterone acetate is being tested in a Phase III trial for men with progressive castration-resistant prostate cancer who are chemotherapy naive . A Phase III trial for patients following prior chemotherapy has been completed and is awaiting analysis . Synthesis , biological evaluation , and molecular modeling of abiraterone analogues : novel P05093 inhibitors for the treatment of prostate cancer . DB05812 , a steroidal cytochrome P450 17alpha-hydroxylase-17,20-lyase inhibitor ( P05093 ) , is currently undergoing phase II clinical trials as a potential drug for the treatment of androgen-dependent prostate cancer . Since steroidal compounds often show side effects attributable to their structure , we have tried to replace the sterane scaffold by nonsteroidal core structures . The design and synthesis of 20 new abiraterone mimetics are described . Their activities have been tested with recombinant human P05093 expressed in E. coli . Promising compounds were further evaluated for selectivity against P15538 , P19099 , and the hepatic P08684 . Compounds 19 and 20 showed comparable activity to abiraterone ( IC50 values of 144 and 64 nM vs 72 nM ) and similar or even better selectivity against the other CYP enzymes . Selected compounds were also docked into our homology model , and the same binding modes as for abiraterone were found . Changing paradigms in management of metastatic Castration Resistant Prostate Cancer ( mCRPC ) . Recently , the standard of care for metastatic Castration Resistant Prostate Cancer ( mCRPC ) has changed considerably . Persistent androgen receptor ( AR ) signaling has been identified as a target for novel therapies and reengages the fact that AR continues to be the primary target responsible for metastatic prostate cancer . P10275 gene amplification and over expression have been found to result in a higher concentration of androgen receptors on tumor cells , making them extremely sensitive to low levels of circulating androgens . Additionally , prostate cancer cells are able to maintain dihydrotestosterone ( DB02901 ) concentration in excess of serum concentrations to support tumor growth . For many years ketoconazole was the only P05093 inhibitor that was used to treat mCRPC . However , significant toxicities limit its use . Newly approved chemotherapeutic agents such as DB05812 ( an oral selective inhibitor of CYP17A ) , which blocks androgen biosynthesis both within and outside the prostate cancer cells ) , and enzalutamide ( blocks AR signaling ) have improved overall survival . There are also ongoing phase III trials for Orteronel ( P50750 - 700 ) , ARN- 509 and Galeterone ( TOK-001 ) , which targets androgen signaling . In this review , we will present the rationale for the newly approved hormonal treatments , their indications and complications , and we will discuss ongoing trials that are being done to improve the efficacy of the approved agents . Finally , we will talk about the potential upcoming hormonal treatments for mCRPC . [ Mechanisms of resistance to P05093 inhibitors in castrate resistant prostate cancer ] . INTRODUCTION : DB05812 acetate has increased the overall survival of patients with metastatic castration-resistant prostate cancer . However , despite an initial response to treatment , many patients develop resistance to the drug . In this paper we present different hypotheses that may explain the emergence of resistance . METHOD : This review was conducted from the PubMed database . The most relevant articles were selected and analyzed . RESULTS : The molecular mechanisms of resistance to abiraterone acetate remain largely elusive . We detailed some of them including the reactivation of the androgen receptor through alternative biosynthesis of androgens , over expression or mutation of the androgen receptor gene , or the action of co-activators . The over expression of P05093 or the alteration of other genes ' expression involved in steroidogenesis could also contribute to the resistance . CONCLUSION : Some of the molecular mechanisms involved in the resistance to abiraterone acetate were detailed . Better understanding of these mechanisms is a key step to allow the emergence of new therapeutic options and personalized treatments of castration resistant prostate cancer . Recent progress in pharmaceutical therapies for castration-resistant prostate cancer . Since 2010 , six drugs have been approved for the treatment of castration-resistant prostate cancer , i.e. , P05093 inhibitor DB05812 , androgen receptor antagonist DB08899 , cytotoxic agent DB06772 , vaccine Sipuleucel-T , antibody DB06643 against receptor activator of nuclear factor kappa B ligand and radiopharmaceutical Alpharadin . All these drugs demonstrate improvement on overall survival , expect for DB06643 , which increases the bone mineral density of patients under androgen deprivation therapy and prolongs bone-metastasis-free survival . Besides further P05093 inhibitors ( Orteronel , Galeterone , VT-464 and CFG920 ) , androgen receptor antagonists ( ARN-509 , ODM-201 , AZD-3514 and EZN-4176 ) and vaccine Prostvac , more drug candidates with various mechanisms or new indications of launched drugs are currently under evaluation in different stages of clinical trials , including various kinase inhibitors and platinum complexes . Some novel strategies have also been proposed aimed at further potentiation of antitumor effects or reduction of side effects and complications related to treatments . Under these flourishing circumstances , more investigations should be performed on the optimal combination or the sequence of treatments needed to delay or reverse possible resistance and thus maximize the clinical benefits for the patients . Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha-reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha-reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer(PCa) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or= +/-1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR; P24385 ) , and signal transduction ( P04626 ; V- P62158 ; Q07889 ) whereas , underexpressed genes ( KLK3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology . Antitumour activity of docetaxel following treatment with the P05093 inhibitor abiraterone : clinical evidence for cross-resistance ? BACKGROUND : DB05812 and docetaxel are both approved treatments for men with metastatic castration-resistant prostate cancer ( mCRPC ) . DB05812 pre-docetaxel is currently undergoing evaluation in a phase III study . In vitro studies indicate that taxanes may act by disrupting androgen receptor signalling . We hypothesised that prior abiraterone exposure would adversely impact docetaxel efficacy . PATIENTS AND METHODS : We retrospectively evaluated activity of docetaxel in mCRPC patients previously treated with abiraterone , using Prostate Cancer Working Group and radiological criteria . RESULTS : Of the 54 patients treated with abiraterone , 35 subsequently received docetaxel . DB01248 resulted in a prostate-specific antigen ( PSA ) decline of ≥50 % in nine patients [ 26 % , 95 % confidence interval ( CI ) 13 % to 43 % ] , with a median time to PSA progression of 4.6 months ( 95 % CI 4.2 % to 5.9 % ) . PSA declines ≥30 % were achieved by 13 patients ( 37 % , 95 % CI 22 % to 55 % ) . The median overall survival was 12.5 months ( 95 % CI 10.6-19.4 ) . All patients who failed to achieve a PSA fall on abiraterone and were deemed abiraterone-refractory were also docetaxel-refractory ( N = 8 ) . In the 24 patients with radiologically evaluable disease , partial responses were reported in four patients ( 11 % ) , none of whom were abiraterone-refractory . CONCLUSION : The activity of docetaxel post-abiraterone appears lower than anticipated and no responses to docetaxel were observed in abiraterone-refractory patients . DB05812 inhibits 3β-hydroxysteroid dehydrogenase : a rationale for increasing drug exposure in castration-resistant prostate cancer . PURPOSE : Treatment with abiraterone ( abi ) acetate prolongs survival in castration-resistant prostate cancer ( CRPC ) . Resistance to abi invariably occurs , probably due in part to upregulation of steroidogenic enzymes and/or other mechanisms that sustain dihydrotestosterone ( DB02901 ) synthesis , which raises the possibility of reversing resistance by concomitant inhibition of other required steroidogenic enzymes . On the basis of the 3β-hydroxyl , Δ(5)-structure , we hypothesized that abi also inhibits 3β-hydroxysteroid dehydrogenase/isomerase ( 3βHSD ) , which is absolutely required for DB02901 synthesis in CRPC , regardless of origins or routes of synthesis . EXPERIMENTAL DESIGN : We tested the effects of abi on 3βHSD activity , androgen receptor localization , expression of androgen receptor-responsive genes , and CRPC growth in vivo . RESULTS : Abi inhibits recombinant 3βHSD activity in vitro and endogenous 3βHSD activity in LNCaP and LAPC4 cells , including conversion of [ (3)H ] -dehydroepiandrosterone ( DB01708 ) to Δ(4)-androstenedione , androgen receptor nuclear translocation , expression of androgen receptor-responsive genes , and xenograft growth in orchiectomized mice supplemented with DB01708 . Abi also blocks conversion of Δ(5)- DB01524 to testosterone by 3βHSD . Abi inhibits 3βHSD1 and 3βHSD2 enzymatic activity in vitro ; blocks conversion from DB01708 to androstenedione and DB02901 with an IC(50) value of less than 1 μmol/L in CRPC cell lines ; inhibits androgen receptor nuclear translocation ; expression of O15393 , prostate-specific antigen , and Q13451 ; and decreases CRPC xenograft growth in DB01708 -supplemented mice . CONCLUSIONS : We conclude that abi inhibits 3βHSD-mediated conversion of DB01708 to active androgens in CRPC . This second mode of action might be exploited to reverse resistance to P05093 inhibition at the standard abi dose by dose-escalation or simply by administration with food to increase drug exposure . Recent findings in the genetics of blood pressure and hypertension traits . We provide an overview of ongoing discovery efforts in the genetics of blood pressure ( BP ) and hypertension ( HTN ) traits . Two large genome-wide association meta-analyses of individuals of European descent were recently published , revealing ~13 new loci for BP traits . Only two of these loci harbor genes in a pathway known to affect BP ( P05093 and P01160 / P16860 ) . Functional variants in these loci are still unknown . Few genome-wide association studies ( GWAS ) of complex diseases have been published from non-European populations . The study of populations with different evolutionary history and linkage disequilibrium ( LD ) structure , such as individuals of African ancestry , may provide an opportunity to further narrow these regions to identify the causal gene(s) . Several collaborative efforts toward discovery of low-frequency variants and copy number variation for BP traits are currently underway . As evidence for new loci for complex diseases accumulates the assessment of the epidemiologic architecture of these variants in populations assumes higher priority . The impact of public health-relevant contexts such as diet , physical activity , psychosocial factors , and aging has not been examined for most common variants associated with BP . Use of prednisone with abiraterone acetate in metastatic castration-resistant prostate cancer . DB05812 acetate , a prodrug of the P05093 inhibitor abiraterone that blocks androgen biosynthesis , is approved for treatment of patients with metastatic castration-resistant prostate cancer ( mCRPC ) in combination with prednisone or prednisolone 5 mg twice daily . This review evaluates the basis for the effects of prednisone on mineralocorticoid-related adverse events that arise because of P05093 inhibition with abiraterone . Coadministration with the recommended dose of glucocorticoid compensates for abiraterone-induced reductions in serum cortisol and blocks the compensatory increase in adrenocorticotropic hormone seen with abiraterone . Consequently , 5 mg prednisone twice daily serves as a glucocorticoid replacement therapy when coadministered with abiraterone acetate , analogous to use of glucocorticoid replacement therapy for certain endocrine disorders . We searched PubMed to identify safety concerns regarding glucocorticoid use , placing a focus on longitudinal studies in autoimmune and inflammatory diseases and cancer . In general , glucocorticoid-related adverse events , including bone loss , immunosuppression , hyperglycemia , mood and cognitive alterations , and myopathy , appear dose related and tend to occur at doses and/or treatment durations greater than the low dose of glucocorticoid approved in combination with abiraterone acetate for the treatment of mCRPC . Although glucocorticoids are often used to manage tumor-related symptoms or to prevent treatment-related toxicity , available evidence suggests that prednisone and dexamethasone might also offer modest therapeutic benefit in mCRPC . Given recent improvements in survival achieved for mCRPC with novel agents in combination with prednisone , the risks of these recommended glucocorticoid doses must be balanced with the benefits shown for these regimens . Antitumor activity with P05093 blockade indicates that castration-resistant prostate cancer frequently remains hormone driven . DB05812 acetate is a potent , selective , and orally bioavailable small molecule inhibitor of P05093 , an enzyme that catalyzes two key serial reactions ( 17 alpha hydroxylase and 17,20 lyase ) in androgen and estrogen biosynthesis . Clinical trials have confirmed that specific inhibition of P05093 is safe and results in clinically important antitumor activity in up to 70 % of castrate patients with advanced prostate cancer resistant to currently available endocrine therapies . These clinical data indicate that castration-resistant prostate cancer frequently remains hormone dependent and has confirmed that this disease should no longer be described as " hormone resistant or refractory " . Biomarker studies , including the analysis of ETS gene fusion status , on patients treated with abiraterone acetate may allow enrichment of patients with a sensitive phenotype in future studies of therapeutics targeting P05093 . Clinical and biochemical consequences of P05093 inhibition with abiraterone given with and without exogenous glucocorticoids in castrate men with advanced prostate cancer . CONTEXT : DB05812 acetate is a small-molecule cytochrome P450 17A1 ( P05093 ) inhibitor that is active in castration-resistant prostate cancer . OBJECTIVE : Our objective was to determine the impact of abiraterone with and without dexamethasone treatment on in vivo steroidogenesis . DESIGN AND METHODS : We treated 42 castrate , castration-resistant prostate cancer patients with continuous , daily abiraterone acetate and prospectively collected blood and urine before and during abiraterone treatment and after addition of dexamethasone 0.5 mg daily . RESULTS : Treatment with single-agent abiraterone acetate was associated with accumulation of steroids with mineralocorticoid properties upstream of P05093 . This resulted in side effects , including hypertension , hypokalemia , and fluid overload , in 38 of 42 patients that were generally treated effectively with eplerenone . Importantly , serum and urinary androgens were suppressed by more than 90 % from baseline . Urinary metabolites of 17-hydroxypregnenolone and 17-hydroxyprogesterone downstream of 17α-hydroxylase remained unchanged . However , 3α5α-17-hydroxypregnanolone , which can be converted via the backdoor pathway toward 5α-dihydrotestosterone , increased significantly and correlated with levels of the major 5α-dihydrotestosterone metabolite androsterone . In contrast , urinary metabolites of 11-deoxycortisol and active glucocorticoids declined significantly . Addition of dexamethasone to abiraterone acetate significantly suppressed DB01285 and endogenous steroids , including 3α5α-17-hydroxypregnanolone . CONCLUSION : P05093 inhibition with abiraterone acetate is characterized by significant suppression of androgen and cortisol synthesis . The latter is associated with a rise in DB01285 that causes raised mineralocorticoids , leading to side effects and incomplete 17α-hydroxylase inhibition . Concomitant inhibition of 17,20-lyase results in diversion of 17-hydroxyprogesterone metabolites toward androgen synthesis via the backdoor pathway . Addition of dexamethasone reverses toxicity and could further suppress androgens by preventing a rise in substrates of backdoor androgen synthesis . Hormonal therapy for prostate cancer : toward further unraveling of androgen receptor function . Prostate cancer is a major cause of cancer-related death in men . Prostate cancer is an androgen-responsive tumor and the treatment of advanced prostate cancer involves hormonal therapy . First-line treatment for advanced prostate cancer is androgen deprivation therapy ( ADT ) , usually with agents that suppress gonadotropins through a pituitary mechanism . DB00644 agonists and antagonists both suppress gonadal release of testosterone , although their activity profiles vary . ADT down-regulates androgen receptor ( AR ) transcriptional activity in the tumor but the response in metastatic disease is transient and tumors progress as castration-resistant prostate cancer ( CRPC ) . Although serum testosterone concentrations decline dramatically with ADT , CRPC growth remains largely dependent on AR activity . Secondary hormonal therapies are then often employed to further dampen AR-driven transcription . These secondary hormonal therapies either further deplete adrenal or intratumoral androgen synthesis , or directly and competitively antagonize AR . New hormonal agents with both of these mechanisms are in clinical trials and show promising activity in patients with CRPC . DB05812 acetate is an inhibitor of P05093 , which is an enzyme required for the synthesis of all androgens and estrogens . MDV3100 is an AR antagonist that has a higher affinity for AR than any other AR antagonist in clinic use . In phase I and phase II clinical trials , both agents have significant activity . These agents and the promise of the development of others provide hope that more effective hormonal therapies may soon be offered to patients , which will improve clinical outcomes . Endothelial cell transforming growth factor-β receptor activation causes tacrolimus-induced renal arteriolar hyalinosis . Arteriolar hyalinosis is a common histological finding in renal transplant recipients treated with the calcineurin inhibitor tacrolimus ; however , the pathophysiologic mechanisms remain unknown . In addition to increasing transforming growth factor ( TGF ) -β levels , tacrolimus inhibits calcineurin by binding to FK506-binding protein 12 ( P62942 ) . P62942 alone also inhibits TGF-β receptor activation . Here we tested whether tacrolimus binding to P62942 removes an inhibition of the TGF-β receptor , allowing ligand binding , ultimately leading to receptor activation and arteriolar hyalinosis . We found that specific deletion of P62942 from endothelial cells was sufficient to activate endothelial TGF-β receptors and induce renal arteriolar hyalinosis in these knockout mice , similar to that induced by tacrolimus . DB00864 -treated and knockout mice exhibited significantly increased levels of aortic TGF-β receptor activation as evidenced by Q15796 /3 phosphorylation , along with increased collagen and fibronectin expression compared to controls . Treatment of isolated mouse aortas with tacrolimus increased TGF-β receptor activation and collagen and fibronectin expression . These effects were independent of calcineurin , absent in endothelial denuded aortic rings , and could be prevented by the small molecule TGF-β receptor inhibitor SB-505124 . Thus , endothelial cell TGF-β receptor activation is sufficient to cause vascular remodeling and renal arteriolar hyalinosis . Inhibition of the androgen receptor by mineralocorticoids at levels physiologically achieved in serum in patients treated with abiraterone acetate . BACKGROUND : DB05812 acetate ( AA ) , a highly potent P05093 inhibitor , has demonstrated marked clinical benefit in patients with metastatic castration-resistant prostate cancer ( CRPC ) . Phase I trials of AA without prednisone showed significant elevation of serum mineralocorticoid concentrations . The aim of this study was to elucidate the biological significance of elevated mineralocorticoid levels on androgen receptor ( AR ) activity in prostate cancer ( PC ) cells . METHODS : Fluorescence resonance energy transfer ( FRET ) assay was used to assess the effect of mineralocorticoids on androgen-induced conformational change of the AR . LAPC4 , LNCaP and LN-AR cells that were cultured and treated with androgens were exposed to mineralocorticoids at varying concentrations , including levels measured in the serum of AA-treated patients in a phase I trial . AR-dependent transcriptional activity and cell growth were measured in these cell lines to determine the biological impact of mineralocorticoids on PC cells . RESULTS : Corticosterone ( CS ) and deoxycorticosterone ( DOC ) inhibited androgen-induced conformational change of the AR in the FRET assay . CS inhibited AR-dependent transcriptional activity and cell growth at concentrations comparable to those measured in the serum of AA-treated patients . DOC inhibited AR transcriptional activity and cell growth at 10-fold greater concentrations than measured in the serum of AA-treated patients . CONCLUSIONS : Mineralocorticoids directly inhibit androgen-induced conformational change of the AR . CS inhibits AR transcriptional activity and PC cell growth at concentrations found in the serum of patients treated with AA . Further investigation of the potential therapeutic implications of mineralocorticoids in AA-treated CRPC patients is warranted . DB05812 acetate , a novel adrenal inhibitor in metastatic castration-resistant prostate cancer . The androgen receptor remains the key player in patients with castration-resistant prostate cancer ( CRPC ) . Available agents capable of blocking early adrenal androgen production have limited activity and can lead to significant toxicities . DB05812 acetate , a pregnenolone analog , is a small molecule that irreversibly inhibits P05093 , a rate-limiting enzyme in androgen biosynthesis . Several clinical trials have demonstrated the safety and efficacy of this compound in men with metastatic CRPC . Recently , a randomized phase 3 trial evaluating abiraterone acetate in docetaxel-refractory CRPC patients demonstrated a survival improvement over placebo-treated patients ( 14.8 vs 10.9 months ; HR 0.646 ; P < 0.0001 ) . A similar trial in the pre-chemotherapy setting has completed accrual and is undergoing analysis . Here we review the rationale and clinical development of abiraterone acetate in men with CRPC . DB05812 acetate : a review of its use in patients with metastatic castration-resistant prostate cancer . DB05812 acetate ( Zytiga(®) ) is an orally administered , selective inhibitor of the 17α-hydroxylase and C17,20-lyase enzymatic activities of cytochrome P450 ( CYP ) 17 . P05093 is required for androgen biosynthesis , with androgen receptor signalling crucial in the progression from primary to metastatic prostate cancer . DB05812 acetate is approved in the European Union and the US , in combination with prednisone or prednisolone , for the treatment of men with metastatic castration-resistant prostate cancer ( CRPC ) . When administered in combination with prednisone in a placebo-controlled , multinational phase III study , abiraterone acetate significantly prolonged overall survival and radiographic progression-free survival ( rPFS ) in men with metastatic CRPC who had previously received docetaxel . In men with metastatic CRPC who had not previously received chemotherapy participating in a placebo-controlled , multinational phase III study , there was a strong trend towards an overall survival benefit , a significant prolongation in rPFS and significant delays in clinical decline , the need for chemotherapy and the onset of pain observed . Given the nature of the therapy , the overall tolerability profile of abiraterone acetate , in combination with prednisone , was acceptable in men with metastatic CRPC . DB05812 acetate is associated with hypokalaemia , hypertension , and fluid retention or oedema , secondary to its mechanism of action , and with cardiac adverse events and hepatotoxicity ; however , in the phase III studies the incidences of the most frequently reported grade 3 or 4 adverse events of special interest were relatively low . Although the final overall survival data in men with metastatic CRPC who have not previously received chemotherapy are awaited , current evidence indicates that abiraterone acetate is a useful option for the treatment of metastatic CRPC . Human heterochromatin protein 1 isoforms regulate androgen receptor signaling in prostate cancer . P10275 ( AR ) signaling is critical for the tumorigenesis and development of prostate cancer , as well as the progression to castration-resistant prostate cancer . We previously showed that the heterochromatin protein 1 ( P59665 ) β isoform plays a critical role in transactivation of AR signaling as an AR coactivator that promotes prostate cancer cell proliferation . However , the roles of other P59665 isoforms , HP1α and HP1γ , in AR expression and prostate cancer remain unclear . Here , we found that knockdown of HP1γ , but not HP1α , reduced AR expression and cell proliferation by inducing cell cycle arrest at P55008 phase in LNCaP cells . Conversely , overexpression of full-length HP1α and its C-terminal deletion mutant increased AR expression and cell growth , whereas overexpression of HP1γ had no effect . Similarly , HP1α overexpression promoted 22Rv1 cell growth , whereas HP1γ knockdown reduced the proliferation of CxR cells , a castration-resistant LNCaP derivative . Taken together , P59665 isoforms distinctly augment AR signaling and cell growth in prostate cancer . Therefore , silencing of HP1β and HP1γ may be a promising therapeutic strategy for treatment of prostate cancer . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . A-ring modified steroidal azoles retaining similar potent and slowly reversible P05093 inhibition as abiraterone . DB05812 acetate is a potent inhibitor of human cytochrome P450c17 ( P05093 , 17α-hydroxylase/17,20-lyase ) and is clinically used in combination with prednisone for the treatment of castration-resistant prostate cancer . Although many studies have documented the potency of abiraterone ( Abi ) in a variety of in vitro and in vivo systems for several species , the exact potency of Abi for human P05093 enzyme has not yet been determined , and the structural requirements for high-potency steroidal azole inhibitors are not established . We synthesized 4 Abi analogs differing in the A-B ring substitution patterns : 3α-hydroxy-Δ(4)-Abi ( 13 ) , 3-keto-Δ(4)-Abi ( 11 ) , 3-keto-5α-Abi ( 6 ) , and 3α-hydroxy-5α-Abi ( 5 ) . We measured the spectral binding constants ( Ks ) using purified and modified human P05093 along with the determination constants ( Ki ) applying a native human P05093 enzyme in yeast microsomes for these compounds as well as for ketoconazole . For Abi , 3-keto-Δ(4)-Abi , 3-keto-5α-Abi , and 3α-hydroxy-5α-Abi , the type 2 spectral changes gave the best fit for a quadratic equation , since in these experiments Ks values were 0.1-2.6nM , much lower than that for ketoconazole and 3α-hydroxy-Δ(4)-Abi ( Ks values were 140 and 1660nM , respectively ) . Inhibition experiments showed mixed inhibition patterns with Ki values of 7-80nM . Abi dissociation from the P05093 -Abi complex was incomplete and slow ; the t1/2 for dissociation was 1.8h , with 55 % of complex remaining after 5h . We conclude that Abi and the 3 related steroidal azoles ( 3-keto-Δ(4)-Abi , 3-keto-5α-Abi , and 3α-hydroxy-5α-Abi ) , which also mimic natural substrates , are extraordinarily potent inhibitors of human P05093 , whereas the 3α-hydroxy-Δ(4)-Abi is moderately potent and comparable to ketoconazole . [ Roles of medical oncologists in the new era of CRPC therapy in Japan ] . Currently , the standard therapy for advanced prostate cancer is endocrine therapy ( luteinizing hormone-releasing hormone [LH-RH]agonists alone or LH-RHagonists plus antiandrogens ) . However , most patients eventually become resistant to these therapies as well as castration therapy . New endocrine therapies for castration-resistant prostate cancer(CRPC)have been developed . DB05812 , a P05093 inhibitor , and enzalutamide , a novel androgen receptor antagonist , have been shown to improve the overall survival , and they are set to be approved in Japan soon . Moreover , docetaxel and cabazitaxel have been established as first- and second-line chemotherapeutic drugs , respectively . Although there is currently no established molecular target drug for CRPC , some drugs such as cabozantinib seem to be effective , and they may be used in the future . In these situations of new drug development , the contribution of medical oncologists is predictable . While medical oncologists can not play central roles in all aspects of drug therapy for urological malignancies in Japan , they must play roles in certain aspects such as new drug development starting from phase I trials , improving multidisciplinary care for adverse events , and promoting translational research . DB05812 acetate , a first-in-class P05093 inhibitor , establishes a new treatment paradigm in castration-resistant prostate cancer . P05093 inhibitors -- abiraterone , C17,20-lyase inhibitors and multi-targeting agents . As the first in class steroid 17α-hydroxylase/C17,20-lyase ( P05093 ) inhibitor , abiraterone acetate ( of which the active metabolite is abiraterone ) has been shown to improve overall survival in patients with castration-resistant prostate cancer ( CRPC ) -- in those who are chemotherapy-naive and those previously treated with docetaxel . Furthermore , the clinical success of abiraterone demonstrated that CRPC , which has previously been regarded as an androgen-independent disease , is still driven , at least in part , by androgens . More importantly , abiraterone is a ' promiscuous ' drug that interacts with a number of targets , which dictate its clinical benefits and adverse effects profile . Besides P05093 inhibition , abiraterone acts as an antagonist to the androgen receptor and inhibits 3β-hydroxysteroid dehydrogenase -- two effects that potentially contribute to its antitumour effects . However , the inhibition of the 17α-hydroxylase activity of P05093 , P15538 and a panel of hepatic CYP enzymes leads to adverse effects and toxicities that include secondary mineralocorticoid excess . DB05812 is also associated with increased incidence of cardiac disorders . Under such circumstances , development of new P05093 inhibitors as an additional line of defence is urgently needed . To achieve enhanced clinical benefits , new strategies are being explored that include selective inhibition of the C17,20-lyase activity of P05093 and multi-targeting strategies that affect androgen synthesis and signalling at different points . Some of these strategies-including the drugs orteronel , VT-464 and galeterone -- are supported by preclinical data and are being explored in the clinic . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . New agents and strategies for the hormonal treatment of castration-resistant prostate cancer . IMPORTANCE OF THE FIELD : Hormonal therapy with medical or surgical castration is the mainstay of systemic therapy for advanced prostate cancer . Depletion of gonadal testosterone in circulation is typically initially effective , although responses are transient and metastatic disease progresses as castration-resistant prostate cancer ( CRPC ) . AREAS COVERED IN THIS REVIEW : CRPC is accompanied by a gain of function in the androgen receptor ( AR ) , which may occur at the level of AR itself or through intratumoral repletion of androgens that in turn stimulate AR . Investigational drugs in clinical trials have promising activity in CRPC . DB05812 acetate is a P05093 inhibitor that blocks the synthesis of adrenal androgens . MDV3100 is a nonsteroidal AR antagonist with a greater binding affinity than other AR antagonists currently in clinical use . Insights into the mechanisms of intratumoral steroidogenesis in CRPC have defined other potential targets . Metabolism from DB01708 to testosterone and dihydrotestosterone requires 3-hydroxyl oxidation and Delta(5) isomerization to Delta(4) by 3beta-hydroxysteroid dehydrogenase ( 3betaHSD ) and 17-keto reduction by 17beta-hydroxysteroid dehydrogenase ( 17betaHSD ) -3 or -5 . AR activation in CRPC by intratumoral steroids requires these enzymatic steps . Investigation into specific inhibitors of 3betaHSD and 17betaHSD are required to determine their efficacy and potential roles in the treatment of CPRC . WHAT THE READER WILL GAIN : Readers will gain an understanding of the biology of CRPC , new investigational hormonal agents and novel approaches to the treatment of CRPC . TAKE HOME MESSAGE : Intratumoral androgens drive CRPC progression . New investigational hormonal agents that inhibit intratumoral androgens are highly active in the treatment of CRPC . Alternative strategies hold the promise for the development of other agents with novel mechanisms of action . Battle of the kinases : integration of adrenal responses to DB02527 , DG and Ca2+ at the level of steroidogenic cytochromes P450 and 3betaHSD expression in H295R cells . While DB01285 receptors ( activating the protein kinase A pathway ) are expressed throughout the human/bovine/ovine zona glomerulosa ( zg ) and zona fasciculata ( zf ) , there are clear zonal differences in AII Type-1 receptor levels ( activating protein kinase C/Ca2+ ) , as well as resting membrane potential . Thus zg is most responsive to AII and K+ ( Ca2+ signalling ) , while zf is less responsive to AII with no K+ response . Zonal function in turn requires differential expression of P05093 /3betaHSD and P19099 / P15538 . We have used the H295R cell to study how differential activation of kinase A , kinase C and Ca2+/calmodulin ( P62158 ) kinases may alter the relative expression of the steroidogenic P450s and 3betaHSDII . While P05108 , P05093 , 3betaHSDII , P08686 , and P15538 are all induced by increases in DB02527 , studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively ( P08686 , 3betaHSDII ) or negatively ( P05093 , P05108 ) by protein kinase C . Thus adrenal 3betaHSDII and P08686 expression is high in zg and zf , but P05093 is not expressed in the zg where AII activation of kinase C is highest . In turn both K+ and AII-induced elevation of Ca2+ strongly induces P19099 but not P15538 , consistent with preferential expression of P19099 in the zg . We conclude that differential signaling through kinase C and P62158 kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones . DB05812 acetate : redefining hormone treatment for advanced prostate cancer . Prostate cancer has long since been recognised as being hormonally driven via androgen receptor signalling . DB05812 acetate ( AA ) is a rationally designed P05093 inhibitor that blocks the conversion of androgens from non-gonadal precursors effectively , thus reducing testosterone to undetectable levels . AA has recently been proved to extend survival for men with metastatic castration-resistant prostate cancer who have progressive disease after first-line chemotherapy treatment . In addition , it is currently being tested in a Phase III trial in the pre-chemotherapy setting . This paper will review the preclinical discovery and clinical development of AA and will outline the strategy of parallel translational research . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB05812 acetate : oral androgen biosynthesis inhibitor for treatment of castration-resistant prostate cancer . Prostate cancer is the second leading cause of cancer death in men in the US and Europe . The treatment of advanced-stage prostate cancer has been androgen deprivation . Medical castration leads to decreased production of testosterone and dihydrotestosterone by the testes , but adrenal glands and even prostate cancer tissue continue to produce androgens , which eventually leads to continued prostate cancer growth despite castrate level of androgens . This stage is known as castrate-resistant prostate cancer ( CRPC ) , which continues to be a challenge to treat . Addition of androgen antagonists to hormonal deprivation has been successful in lowering the prostate-specific antigen levels further , but has not actually translated into life-prolonging options . The results of several contemporary studies have continued to demonstrate activation of the androgen receptor as being the key factor in the continued growth of prostate cancer . Blockade of androgen production by nongonadal sources has led to clinical benefit in this setting . One such agent is abiraterone acetate , which significantly reduces androgen production by blocking the enzyme , cytochrome P450 17 alpha-hydroxylase ( P05093 ) . This has provided physicians with another treatment option for patients with CRPC . The landscape for prostate cancer treatment has changed with the approval of cabazitaxel , sipuleucel-T and abiraterone . Here we provide an overview of abiraterone acetate , its mechanism of action , and its potential place for therapy in CRPC . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . Retreatment of men with metastatic castrate-resistant prostate cancer with abiraterone . BACKGROUND : DB05812 acetate ( AA ) , oral P05093 inhibitor , is an active agent in the treatment of metastatic castrate-resistant prostate cancer ( mCRPC ) . METHODS : We ( R.L.A and N.A ) retrospectively evaluated outcome in 12 men who were re-treated with AA following prior treatment with AA at the Princess Margaret Cancer Centre . RESULTS : All men were heavily pre-treated for mCRPC with a median of four prior lines of therapy , one of which was AA ( given either pre- or post-chemotherapy ) . Eleven out of 12 ( 92 % ) men stopped their first treatment course of AA due to progression and one stopped for financial reasons . Seven men had a PSA decrease ≥50 % following their first AA treatment , of which three ( 46 % ) had a PSA decrease ≥50 % to AA re-treatment . The responses to AA re-treatment were generally short-lived with a median biochemical progression-free survival of 2.3 months and median treatment duration of 3.2 months . No PSA responses to AA re-treatment were seen in five men who did not have an initial PSA response to AA . CONCLUSIONS : Our data suggest that AA re-challenge may have limited benefit in select men with mCRPC , and warrants further formal research . Inhibin removes the inhibitory effects of activin on steroid enzyme expression and androgen production by normal ovarian thecal cells . Activin and inhibin are important local modulators of theca cell steroidogenesis in the ovary . Using a serum-free primary theca cell culture system , this study investigated the effects of inhibin on theca cell androgen production and expression of steroidogenic enzymes . Androstenedione secretion from theca cells cultured in media containing activin , inhibin and follistatin was assessed by RIA over 144 h . Activin ( 1-100 ng/ml ) suppressed androstenedione production . Inhibin ( 1-100 ng/ml ) blocked the suppressive effects of added activin , but increased androstenedione production when added alone , suggesting it was blocking endogenous activin produced by theca cells . Addition of SB-431542 ( activin receptor inhibitor ) and follistatin ( 500 ng/ml ) increased androstenedione production , supporting this concept . Infection of theca cells with adenoviruses expressing inhibitory O43541 or 7 increased androstenedione secretion , confirming that the suppressive effects of activin required activation of the Q15796 /3 pathway . Activin decreased the expression levels of steroidogenic acute regulatory protein ( STAR ) , whereas STAR expression was increased by inhibin and SB-431542 , alone and in combination . P05108 was unaffected . The expression of P05093 encoding 17α-hydroxylase was unaffected by activin but increased by inhibin and SB-431542 , and when added in combination the effect was further enhanced . The expression of 3β-hydroxysteroid dehydrogenase ( 3β-HSD ) was significantly decreased by activin , while inhibin alone and in combination with SB-431542 both potently increased the expression of 3β-HSD . In conclusion , activin suppressed theca cell androstenedione production by decreasing the expression of STAR and 3β-HSD . Inhibin and other blockers of activin action reversed this effect , supporting the concept that endogenous thecal activin modulates androgen production in theca cells . [ DB05812 acetate(ZYTIGA®)-development and literature review ] . DB05812 acetate(AA)has been approved in more than 80 countries for the treatment of patients with metastatic castration-resistant prostate cancer(mCRPC) . In July 2013 , a marketing approval application for AA was submitted to the Japanese Ministry of Health , Labour , and Welfare . AA is a selective inhibitor of P05093 , a crucial enzyme for androgen biosynthesis . AA exerts its anti-tumor activity by directly inhibiting androgen production at all three sources , i. e. , the testes , adrenal glands , and tumor itself . Data from international phase III studies and phase I and II studies in Japan have indicated that AA improves the overall survival and quality of life(QoL)of patients with mCRPC . Herein , we have summarized the development of AA and the results of important international and local clinical trials in Japan . In addition , the effect of food on AA bioavailability , concomitant steroid use , and liver function test abnormalities have been discussed regarding the appropriate use of AA .

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interacts_with DB08870?

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Serum macrophage migration inhibitory factor ( MIF ) levels after allogeneic hematopoietic stem cell transplantation . P14174 ( MIF ) may play an important role in the pathogenesis of acute graft-versus-host disease ( aGVHD ) after allogeneic hematopoietic stem cell transplantation ( allo-HSCT ) , as MIF plays an important role to regulate the production of tumor necrosis factor-alpha ( P01375 ) , one of the inflammatory cytokines which induces and exacerbates aGVHD . We examined the association between serum MIF levels and aGVHD vs. chronic GVHD ( cGVHD ) in allo- P09683 patients in this study . We found a significant increase in the peak serum MIF ( 14.46 ng +/- 1.47 ng/ml ) at onset in patients that developed aGVHD ( n = 23 , P = 0.009 ) . We also found that mean serum MIF levels in patients who developed extensive type cGVHD within 6 months ( 12.58 +/- 2.18 ng/ml , n = 13 ) were significantly higher than MIF levels before allo-HSCT ( 7.86 +/- 1.17 ng/ml , n = 19 , P = 0.04 ) . Therefore , we speculated that serum MIF levels increase during the active phase of both aGVHD and cGVHD . Enhanced vasoactive intestinal peptide-induced prolactin secretion from anterior pituitary cells of incubating turkeys ( Meleagris gallopavo ) . During incubation , female turkeys exhibit elevated circulating prolactin ( PRL ) which may be the result of enhanced pituitary responsiveness to vasoactive intestinal peptide ( P01282 ) . This hypothesis was tested by comparison of spontaneous and porcine P01282 -induced PRL secretion from anterior pituitary cells of hens in various reproductive conditions . The effect of P01282 and luteinizing hormone releasing hormone ( P01148 ) , alone and in combination , on luteinizing hormone ( LH ) secretion was also examined . Incubation with pVIP ( 10(-10) to 10(-6) M ) significantly stimulated PRL secretion at all incubation times tested ( 1-5 hr ) . This increase was greatest in cells from incubating hens , with those from laying , photorefractory , and quiescent ( nonphotostimulated ) hens secreting successively less PRL . These responses were obtained when spontaneous PRL secretions were compared . P01282 induced approximately a similar 1.5-fold increase in LH secretion , in all reproductive groups . Also , P01282 enhanced P01148 -induced LH secretion ( 1.2- to 1.6-fold ; P less than 0.0001 ) . It is concluded that PRL secretion in vitro by pituitary cells from turkey hens in various reproductive stages reflects the circulating levels of PRL at these stages . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . G- P04141 downregulates natural killer cell-mediated cytotoxicity in donors for hematopoietic P09683 . In G- P04141 -mobilized hematopoietic P09683 ( HSCT ) , natural killer ( NK ) cells have a critical role in GVHD and GVL effects . However , regulation of NK cell response to G- P04141 remains unclear . This study assayed G- P04141 effects in both HSCT donors and NK-92MI cells . The donors who received G- P04141 had significantly decreased NK cell cytotoxicity . Levels of phosphatidylinositol 3-kinase ( PI3K ) and phosphorylated (p)-Akt , but not mammalian target of rapamycin ( P42345 ) , were downregulated in NK cells from G- P04141 -injected donors . G- P04141 also decreased cytotoxicity without affecting viability and NF-κB of NK-92MI cells . PI3K and p- P29323 expression were also decreased in G- P04141 -treated NK-92MI cells , and their inhibitors , wortmannin and PD98059 , respectively , both enhanced the downregulation of cytotoxicity . These effects were accompanied by decreased expression of a cytotoxicity-related gene , triosephosphate isomerase ( P60174 ) . Wortmannin , but not PD98059 , enhanced the downregulation of P60174 in G- P04141 -treated NK-92MI cells , indicating a correlation between PI3K and P60174 . We conclude that G- P04141 -impaired NK cell cytotoxicity may accompany PI3K/Akt signaling . The effect is transient and NK cells may recover after G- P04141 clearance , suggesting that G- P04141 -mobilized HSCT may benefit both acute GVHD prevention and late-phase GVL promotion in HSCT recipients . IGF-IR tyrosine kinase interacts with P06748 - Q9UM73 oncogene to induce survival of T-cell Q9UM73 + anaplastic large-cell lymphoma cells . Type I insulin-like growth factor receptor ( IGF-IR ) tyrosine kinase plays important roles in the pathogenesis of several malignancies . Although it promotes the growth of stimulated hematopoietic cells , a direct role of IGF-IR in malignant lymphoma has not been identified . Q9UM73 -positive anaplastic large-cell lymphoma ( Q9UM73 (+) ALCL ) is a unique type of T-cell lymphoma . Approximately 85 % of Q9UM73 (+) ALCL cases harbor the translocation t(2;5)(p23;q35) , which generates the chimeric oncogene P06748 - Q9UM73 . In the present study , we explored a possible role of IGF-IR in Q9UM73 (+) ALCL . Our results demonstrate that IGF-IR and P05019 are widely expressed in Q9UM73 (+) ALCL cell lines and primary tumors . Importantly , we identified novel reciprocal functional interactions between IGF-IR and P06748 - Q9UM73 . Antagonism of IGF-IR decreased the viability , induced apoptosis and cell-cycle arrest , and decreased proliferation and colony formation of Q9UM73 (+) ALCL cell lines . These effects could be explained by alterations of cell survival regulatory proteins downstream of IGF-IR signaling . Our findings improve current understanding of the biology of IGF-IR and P06748 - Q9UM73 and have significant therapeutic implications as they identify IGF-IR signaling as a potential therapeutic target in Q9UM73 (+) ALCL and possibly other types of malignant lymphoma . Suppression of experimental autoimmune encephalomyelitis using peptide mimics of P10747 . The P33681 : P10747 / P16410 costimulatory pathway plays a critical role in regulating the immune response and thus provides an ideal target for therapeutic manipulation of autoimmune disease . Previous studies have shown that blockade of P10747 signaling by mAbs can both prevent and exacerbate experimental autoimmune encephalomyelitis ( EAE ) . In this study , we have designed two P10747 peptide mimics that selectively block P33681 : P10747 interactions . By surface plasmon resonance , both the end group-blocked P10747 peptide ( EL- P10747 ) and its retro-inverso isomer ( RI- P10747 ) compete effectively with the extracellular domain of P10747 for binding to P33681 -1 . Both the P10747 peptide mimics inhibited expansion of encephalitogenic T cells in vitro . A single administration of EL- P10747 or RI- P10747 peptide significantly reduced disease severity in EAE . Importantly , we show that either P10747 peptide mimic administered during acute disease dramatically improved clinical signs of EAE , suppressing ongoing disease . The ratio of P33681 : P42081 expression was significantly lower on P01730 (+) and F4/80(+) spleen cells in P10747 peptide-treated mice . Peripheral deletion of Ag-specific P01730 (+) T cells occurs following in vivo blockade of P10747 with synthetic P10747 peptides . An assessment of the effect of human herpesvirus-6 replication on active cytomegalovirus infection after allogeneic stem cell transplantation . Human herpesvirus-6 ( HHV-6 ) may enhance cytomegalovirus ( CMV ) replication in allogeneic stem cell transplant ( allo- P09683 ) recipients either through direct or indirect mechanisms . Definitive evidence supporting this hypothesis are lacking . We investigated the effect of HHV-6 replication on active CMV infection in 68 allo- P09683 recipients . Analysis of plasma HHV-6 and CMV DNAemia was performed by real-time PCR . Enumeration of pp65 and IE-1 CMV-specific IFNgamma CD8(+) and P01730 (+)T cells was performed by intracellular cytokine staining . HHV-6 DNAemia occurred in 39.8 % of patients , and was significantly associated with subsequent CMV DNAemia in univariate ( P=.01 ) , but not in multivariate analysis ( P=.65 ) . The peak of HHV-6 DNAemia was not predictive of the development of CMV DNAemia . Timing and kinetics of active CMV infection were comparable in patients either with or without a preceding episode of HHV-6 DNAemia . The occurrence of HHV-6 DNAemia had no impact on CMV-specific T cell immunity reconstitution early after transplant . The receipt of a graft from an HLA-mismatched donor was independently associated with HHV-6 ( P=.009 ) and CMV reactivation ( P=.04 ) . The data favor the hypothesis that a state of severe immunosuppression leads to HHV-6 and CMV coactivation , but argue against a role of HHV-6 in predisposing to the development of CMV DNAemia or influencing the course of active CMV infection . Immunoreceptor tyrosine-based inhibitory motifs on activating molecules . Immunoreceptor tyrosine-based inhibitory motifs ( ITIMs ) have the restricted consensus sequence V/I/xYxxL/V , but may be more broadly defined by the sequence V/I/L/SxYxxL/V/I/S . If one includes the ITIM of P16410 , then the sequence becomes psixYxxpsi , where psi represents amino acids with nonpolar side chains . Aside from their presence in various inhibitory molecules , ITIMs are also found on many activating receptors and pathways . ITIMs with the restricted consensus sequence occur on IL-4Ralpha , IL-3Rbeta type II , P40189 cytokineR , P48357 ( leptinR ) , P15018 -Rbeta P19438 , Q99062 , PDGF-R , Blk , Ctk/Ntk , Lsk , Zap-70 , P31749 /RACalpha , P17252 , P05771 , P05129 , PKC-delta , PKC-zeta , PKC-epsilon , PKC-eta , PKC-phi , PKC-mu , calmodulin-dependent kinase IIdelta , SLP-76-associated protein , O15117 , Shc binding protein , RasGRF2 , Q13972 homologue , Jak2 , Jak3 , PLCbeta1 , and PLCbeta3 . If ITIMs are defined by a broader consensus sequence , the list of ITIMs on activating molecules becomes even larger . In some instances , these ITIMs have been shown to associate with inhibitory phosphatases . Whether these ITIMs on activating receptors/pathways are necessary and sufficient for negative control of activating events and for immunologic tolerance is not yet known . In some instances , ITIMs on coinhibitory receptors are also required for appropriate negative regulation . By studying events leading to negative control during activation and to immunologic tolerance , it should be possible to discern the balance between antigen receptor-based negative events and coinhibition . DB08870 : its role in the treatment of anaplastic large cell and Hodgkin 's lymphoma . DB08870 is being developed in a joint collaboration between Seattle Genetics and Millennium : The Takeda Oncology Company . In August 2011 , it was approved by the FDA for the treatment of patients with Hodgkin 's lymphoma ( HL ) and anaplastic large cell lymphoma ( ALCL ) . DB08870 is an antibody-drug conjugate that specifically targets the P01375 receptor superfamily member 8 ( P28908 ) antigen on the surface of cancer cells to induce cell death . DB08870 has shown efficacy in inducing apoptosis in HL and ALCL cell lines that express P28908 and reducing tumor size in preclinical models . DB08870 is under clinical evaluation for the treatment of relapsed or refractory HL and ALCL in both adults and children . It is being investigated for use as a combination agent with pre-existing frontline chemotherapies and as a stand-alone salvage therapy for use prior to autologous stem cell transplant . Treatment with brentuximab vedotin is generally well tolerated although it is associated with grade 1-2 adverse reactions such as neutropenia and there have been reports of grade 3-4 serious adverse events . In particular its use with chemotherapy regimens that include bleomycin is contraindicated because of adverse pulmonary effects . Regulation of P33681 .1 costimulatory molecule is mediated by the IFN regulatory factor-7 through the activation of JNK in lipopolysaccharide-stimulated human monocytic cells . The engagement of P10747 or P16410 with P33681 .1 provides the essential second costimulatory signal that regulates the development of immune responses , including T cell activation , differentiation , and induction of peripheral tolerance . The signaling molecules and the transcription factors involved in P33681 .1 regulation are poorly understood . In this study we investigated the role of MAPKs in the regulation of LPS-induced P33681 .1 expression in human monocytes and the promonocytic THP-1 cells . Our results show that LPS-induced P33681 .1 expression in monocytic cells did not involve the activation of either p38 or ERKs . Using the JNK-specific inhibitor SP600125 , small interfering RNAs specific for P45983 and P45984 , and agents such as dexamethasone that inhibit JNK activation , we determined that LPS-induced P33681 .1 expression was regulated by JNK MAPK in both monocytes and THP-1 cells . In addition , we identified a distinct P33681 .1-responsive element corresponding to the IFN regulatory factor-7 ( Q92985 ) binding site in the P33681 .1 promoter responsible for the regulation of LPS-induced P33681 .1 transcription . Furthermore , SP600125 and dexamethasone inhibited LPS-induced Q92985 activity . Taken together , these results suggest that LPS-induced P33681 .1 transcription in human monocytic cells may be regulated by JNK-mediated activation of the Q92985 transcription factor . Hypoxia-microRNA-16 downregulation induces P15692 expression in anaplastic lymphoma kinase ( Q9UM73 ) -positive anaplastic large-cell lymphomas . The anaplastic lymphoma kinase ( Q9UM73 ) , tyrosine kinase oncogene is implicated in a wide variety of cancers . In this study we used conditional onco- Q9UM73 ( P06748 - Q9UM73 and P06753 - Q9UM73 ) mouse MEF cell lines ( Q9UM73 + fibroblasts ) and transgenic models ( Q9UM73 + B-lymphoma ) to investigate the involvement and regulation of angiogenesis in Q9UM73 tumor development . First , we observed that Q9UM73 expression leads to downregulation of miR-16 and increased Vascular Endothelial Growth Factor ( P15692 ) levels . Second , we found that modification of miR-16 levels in P06753 - Q9UM73 MEF cells greatly affected P15692 levels . Third , we demonstrated that miR-16 directly interacts with P15692 mRNA at the 3'-untranslated region and that the regulation of P15692 by miR-16 occurs at the translational level . Fourth , we showed that expression of both the Q9UM73 oncogene and hypoxia-induced factor 1α ( HIF1α ) is a prerequisite for miR-16 downregulation . Fifth , in vivo , miR-16 gain resulted in reduced angiogenesis and tumor growth . Finally , we highlighted an inverse correlation between the levels of miR-16 and P15692 in human P06748 - Q9UM73 + Anaplastic Large Cell Lymphomas ( ALCL ) . Altogether , our results demonstrate , for the first time , the involvement of angiogenesis in Q9UM73 + ALCL and strongly suggest an important role for hypoxia-miR-16 in regulating P15692 translation . Mutated regions of nucleophosmin 1 elicit both P01730 (+) and CD8(+) T-cell responses in patients with acute myeloid leukemia . Mutations in the nucleophosmin gene ( P06748 (mut) ) are one of the most frequent molecular alterations in acute myeloid leukemia ( AML ) , and immune responses may contribute to the favorable prognosis of AML patients with P06748 (mut) . In the present study , we were able to demonstrate both P01730 (+) and CD8(+) T-cell responses against P06748 (mut) . Ten peptides derived from wild-type P06748 and P06748 (mut) were subjected to ELISPOT analysis in 33 healthy volunteers and 27 AML patients . Tetramer assays against the most interesting epitopes were performed and Cr(51)-release assays were used to show the cytotoxicity of peptide-specific T cells . Moreover , HLA-DR-binding epitopes were used to test the role of P01730 (+) T cells in P06748 immunogenicity . Two epitopes ( epitopes # 1 and # 3 ) derived from P06748 (mut) induced CD8(+) T-cell responses . A total of 33 % of the P06748 (mut) AML patients showed immune responses against epitope # 1 and 44 % against epitope # 3 . Specific lysis of leukemic blasts was detected . To obtain robust immune responses against tumor cells , the activation of P01730 (+) T cells is crucial . Therefore , overlapping ( OL ) peptides were analyzed in ELISPOT assays and OL8 was able to activate both CD8(+) and P01730 (+) T cells . The results of the present study show that P06748 (mut) induces specific T-cell responses of P01730 (+) and CD8(+) T cells and therefore is a promising target for specific immunotherapies in AML . VIPhyb , an antagonist of vasoactive intestinal peptide receptor , enhances cellular antiviral immunity in murine cytomegalovirus infected mice . Vasoactive intestinal peptide ( P01282 ) is a neuropeptide hormone that suppresses Th1-mediated cellular immunity . We previously reported that P01282 -knockout ( P01282 -KO ) mice have enhanced cellular immune responses and increased survival following murine cytomegalovirus ( mCMV ) infection in C57BL/6 mice . In this study , we tested whether treatment with a P01282 receptor antagonistic peptide protects C57BL/6 and BALB/c mice from mCMV-infection . One week of daily subcutaneous injections of VIPhyb was non-toxic and did not alter frequencies of immune cell subsets in non-infected mice . VIPhyb administration to mCMV-infected C57BL/6 and BALB/c mice markedly enhanced survival , viral clearance , and reduced liver and lung pathology compared with saline-treated controls . The numbers of effector/memory CD8+ T-cells and mature NK cells were increased in VIPhyb-treated mice compared with PBS-treated groups . Pharmacological blockade of P01282 -receptor binding or genetic blockade of P01282 -signaling prevented the up-regulation of Q9NZQ7 and P18621 expression on DC and activated CD8+ T-cells , respectively , in mCMV-infected mice , and enhanced P33681 , P42081 , and MHC-II expression on conventional and plasmacytoid DC . VIPhyb-treatment increased type-I IFN synthesis , numbers of IFN-γ- and P01375 -α-expressing NK cells and T-cells , and the numbers of mCMV-M45 epitope-peptide-MHC-I tetramer CD8+ T-cells following mCMV infection . P01282 -treatment lowered the percentage of Treg cells in spleens compared with PBS-treated WT mice following mCMV infection , while significantly decreasing levels of serum P15692 induced by mCMV-infection . The mice in all treated groups exhibited similar levels of anti-mCMV antibody titers . Short-term administration of a P01282 -receptor antagonist represents a novel approach to enhance innate and adaptive cellular immunity in a murine model of CMV infection . The role of atorvastatin in regulating the immune response leading to vascular damage in a model of Kawasaki disease . Superantigens have been implicated in a number of diseases including Kawasaki disease ( KD ) , a multi-system vasculitis resulting in coronary artery aneurysms . We have characterized a murine disease model in which coronary arteritis is induced by a novel superantigen found in Lactobacillus casei cell wall extract ( LCWE ) . Using this animal model of KD , we have identified three pathogenic steps leading to coronary artery aneurysm formation . These steps include T cell activation and proliferation , production of the proinflammatory cytokine tumour necrosis factor ( P01375 ) -α and up-regulation of matrix metalloproteinase 9 ( P14780 ) , an elastolytic protease . In addition to their cholesterol-lowering effects , 3-hydroxy-3-methylglutaryl ( HMG ) coenzyme A ( DB01992 ) reductase inhibitors ( statins ) have pleotropic immunomodulatory properties . Thus , we examined the effect of atorvastatin in modulating each of these three critical pathogenic processes leading to aneurysm formation in the disease model . DB01076 inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner . This inhibition was also observed for production of soluble mediators of inflammation including interleukin ( IL ) -2 and P01375 -α . The inhibitory effect on proliferation was rescued completely by mevalonic acid , confirming that the mechanism responsible for this inhibitory activity on immune activation was inhibition of P04035 . Similarly , P01375 -α-induced P14780 production was reduced in a dose-dependent manner in response to atorvastatin . Inhibition of extracellular-regulated kinase ( P29323 ) phosphorylation appears to be the mechanism responsible for inhibition of P14780 production . In conclusion , atorvastatin is able to inhibit critical steps known to be important in the development of coronary aneurysms , suggesting that statins may have therapeutic benefit in patients with KD . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Inhibitors of DB00171 -binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages . DB00171 -binding cassette ( DB01048 ) transporters are a large family of proteins whose role is to translocate various substances across biological membranes . They include the Tangier disease protein ABC1 , sulfonylurea receptors ( Q09428 ) , multidrug resistance protein ( MDR ) , and cystic fibrosis transmembrane regulator ( P13569 ) . In the current study , we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide ( LPS ) and/or interferon ( IFN ) -gamma-induced interleukin ( IL ) -12 p40 and tumor necrosis factor ( P01375 ) -alpha production , nitric oxide formation , as well as major histocompatibility complex II up-regulation in macrophages . The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production . However , glibenclamide failed to affect the production of P01375 . The selective ABC1 inhibitors 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production . On the other hand , both the MDR inhibitor verapamil and P13569 blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40 . Furthermore , selective inhibitors and activators of SURs were without effect . In agreement with the pharmacological data , macrophages expressed mRNA for ABC1 , but not SURs or P13569 . Intracellular levels of IL-12 p40 were decreased by glibenclamide , suggesting that glibenclamide does not affect IL-12 p40 secretion . The effect of glibenclamide did not involve an interference with the activation of the p38 and Q8NFH3 /44 mitogen-activated protein kinases or c-Jun kinase . DB01016 also suppressed P01579 -induced up-regulation of major histocompatibility complex II . Taken together , our results indicate that ABC proteins regulate LPS and/or P01579 -induced macrophage activation . Results of a pivotal phase II study of brentuximab vedotin for patients with relapsed or refractory Hodgkin 's lymphoma . PURPOSE : DB08870 is an antibody-drug conjugate ( ADC ) that selectively delivers monomethyl auristatin E , an antimicrotubule agent , into P28908 -expressing cells . In phase I studies , brentuximab vedotin demonstrated significant activity with a favorable safety profile in patients with relapsed or refractory P28908 -positive lymphomas . PATIENTS AND METHODS : In this multinational , open-label , phase II study , the efficacy and safety of brentuximab vedotin were evaluated in patients with relapsed or refractory Hodgkin 's lymphoma ( HL ) after autologous stem-cell transplantation ( auto- P09683 ) . Patients had histologically documented P28908 -positive HL by central pathology review . A total of 102 patients were treated with brentuximab vedotin 1.8 mg/kg by intravenous infusion every 3 weeks . In the absence of disease progression or prohibitive toxicity , patients received a maximum of 16 cycles . The primary end point was the overall objective response rate ( ORR ) determined by an independent radiology review facility . RESULTS : The ORR was 75 % with complete remission ( CR ) in 34 % of patients . The median progression-free survival time for all patients was 5.6 months , and the median duration of response for those in CR was 20.5 months . After a median observation time of more than 1.5 years , 31 patients were alive and free of documented progressive disease . The most common treatment-related adverse events were peripheral sensory neuropathy , nausea , fatigue , neutropenia , and diarrhea . CONCLUSION : The ADC brentuximab vedotin was associated with manageable toxicity and induced objective responses in 75 % of patients with relapsed or refractory HL after auto- P09683 . Durable CRs approaching 2 years were observed , supporting study in earlier lines of therapy . DB00644 agonists reduce the migratory and the invasive behavior of androgen-independent prostate cancer cells by interfering with the activity of P05019 . Androgen-independent prostate carcinoma is characterized by a high proliferation rate and by a strong metastatic behavior . We have previously shown that DB00644 agonists exert a direct and specific inhibitory action on the proliferation of androgen-independent prostate cancer cells ( DU 145 ) . These compounds mainly act by interfering with the mitogenic activity of growth factors , such as the insulin-like growth factor-I ( P05019 ) . The present experiments were performed to clarify whether DB00644 agonists might also affect the migratory and the invasive behavior of androgen-independent prostate cancer cells and to define their mechanism of action . First we showed that the DB00644 agonist DB00007 reduces the migration of DU 145 cells towards a chemoattractant and their ability to invade a reconstituted basement membrane . Experiments were then performed to clarify whether the DB00644 agonist might act by interfering with the pro-metastatic activity of P05019 . We found that , in androgen-independent prostate cancer cells , DB00007 : a ) interferes with the P05019 system ( receptor protein expression and tyrosine-phosphorylation ) ; b ) abrogates the P05019 -induced phosphorylation of Akt ( a kinase previously shown by us to mediate the pro-metastatic activity of P05019 in prostate cancer cells ) ; c ) counteracts the migration and the invasive activity of the cells stimulated by P05019 ; d ) abolishes the effects of P05019 on cell morphology , on actin cytoskeleton organization and on alphavbeta3 integrin expression/cellular localization . These data indicate that DB00644 agonists , in addition to their well known antiproliferative effect , can also exert a significant inhibitory activity on the migratory and invasive behavior of androgen-independent prostate cancer cells , expressing the P30968 . DB00644 agonists act by interfering with the pro-metastatic activity of the growth factor P05019 . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells . Antibody-drug conjugates ( ADC ) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer . DB08870 represents a first-in-class ADC directed against P28908 (+) malignancies . We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response . In this study , we demonstrate that the dolastatin family of microtubule inhibitors , from which the cytotoxic component of brentuximab vedotin is derived , comprises potent inducers of phenotypic and functional dendritic cell ( DC ) maturation . In addition to the direct cytotoxic effect on tumor cells , dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes . Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells . Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins , the antitumor effect was far less pronounced in immunocompromised mice . We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the P18621 - Q9NZQ7 and P16410 coinhibitory pathways . Ultimately , treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients . Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies , such as brentuximab vedotin , with immune-based therapies . Circulating endothelial progenitor cells decreased in patients with sclerodermatous chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a common late complication of allogeneic stem cell transplantation ( allo- P09683 ) . Some cGVHD patients develop skin lesions , and the skin lesions in sclerodermatous cGVHD ( s-cGVHD ) patients resemble those in progressive systemic sclerosis ( PSS ) , which is characterized by impaired production of circulating endothelial progenitor cells ( EPCs ) . We investigated , retrospectively , whether low EPC production may promote the development of sclerodermatous lesions in cGVHD . Peripheral blood ( PB ) was obtained from 14 healthy volunteers and 27 allo- P09683 patients . Five patients developed s-cGVHD . P28906 (+) cells were purified by using the magnetic cell-sorting separation system , and the P28906 (+)/CD133(+)/vascular endothelial growth factor ( P15692 ) receptor-2(+) EPCs were quantified . The endothelial cell colony-formation potential was evaluated . Serum P15692 and basic fibroblast growth factor ( b-FGF ) concentrations were measured by ELISA . The s-cGVHD patients had significantly lower median circulating EPCs frequencies than non-s-cGVHD patients or control ( 145 of 20 mL [ interquartial range-IQR 107-193 ] versus 1083.5 [ IQR 669.3-2151 ] ; P = .0023 , and versus 1530.5 [ IQR 961.3-2158 ] ; P = .0012 , respectively ) . They also had impaired median endothelial-forming ability compared to non-s-cGVHD patients or controls ( 3.8 [ IQR 1.0-4.3 ] versus 12.8 [ IQR 8.8-28.8 ] , and versus 26.4 [ IQR 23.6-30.6 ] , respectively ; P = .0012 ) . Their P15692 and b-FGF serum levels were also higher than in controls . In conclusion , s-cGVHD patients show findings consistent with those seen in PSS with impaired vasculogenesis that may limit blood perfusion and may contribute to the development of sclerodermatous lesions . DB01016 exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803 . DB01016 , a blocker of DB00171 -sensitive potassium ( K( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC-803 cells expressed the K( DB00171 ) channels composed of Kir6.2 and Q09428 subunits . DB01016 induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC-803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion ( O2- ) generation , and caused mitochondrial respiration dysfunction in MGC-803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2-terminal kinase ( JNK ) in MGC-803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 -/- or P45984 -/- MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer . DB08870 in anaplastic large cell lymphoma . INTRODUCTION : DB08870 , a novel anti- P28908 antibody-drug conjugate , delivers a cytotoxic agent into P28908 (+) cells . P28908 expression is characteristic of anaplastic large cell lymphoma ( ALCL ) and Hodgkin lymphoma ( HL ) . AREAS COVERED : We reviewed data on brentuximab vedotin , focusing on ALCL and discuss pharmacology , clinical trials leading to approval and future research directions . Systemic ALCL , 3 % of adult Q9NZ71 , is characterized by large anaplastic P28908 (+) cells . The fusion protein P06748 - Q9UM73 , when present in systemic ALCL , confers better prognosis , although even Q9UM73 - patients with IPI score ≥ 3 are high-risk . For patients with systemic ALCL , 25 - 45 % relapse after frontline therapy , and > 50 % of patients will relapse following high-dose chemotherapy with autologous stem-cell support . There has been no standard therapy for relapsed/refractory systemic ALCL . DB08870 , combines a monoclonal antibody targeted to P28908 with a microtubule disrupting agent and was recently approved for treatment of patients with systemic ALCL that is refractory or relapsed after at least one multiagent chemotherapy regimen . EXPERT OPINION : DB08870 provides targeted therapy to P28908 (+) lymphomas , including ALCL and HL , with high response rates and manageable toxicity , predominantly myelosuppression and peripheral neuropathy . P06748 - Q9UM73 oncogenic kinase promotes cell-cycle progression through activation of JNK/cJun signaling in anaplastic large-cell lymphoma . Anaplastic large-cell lymphoma ( ALCL ) frequently carries the t(2;5)(p23;q35) , resulting in aberrant expression of nucleophosmin-anaplastic lymphoma kinase ( P06748 - Q9UM73 ) . We show that in 293T and Jurkat cells , forced expression of active P06748 - Q9UM73 , but not kinase-dead mutant P06748 - Q9UM73 ( 210K > R ) , induced JNK and cJun phosphorylation , and this was linked to a dramatic increase in AP-1 transcriptional activity . Conversely , inhibition of Q9UM73 activity in P06748 - Q9UM73 (+) ALCL cells resulted in a concentration-dependent dephosphorylation of JNK and cJun and decreased AP-1 DNA-binding . In addition , JNK physically binds P06748 - Q9UM73 and is highly activated in cultured and primary P06748 - Q9UM73 (+) ALCL cells . cJun phosphorylation in P06748 - Q9UM73 (+) ALCL cells is mediated by JNKs , as shown by selective knocking down of P45983 and P45984 genes using siRNA . Inhibition of JNK activity using SP600125 decreased cJun phosphorylation and AP-1 transcriptional activity and this was associated with decreased cell proliferation and G2/M cell-cycle arrest in a dose-dependent manner . Silencing of the cJun gene by siRNA led to a decreased S-phase cell-cycle fraction associated with upregulation of P38936 and downregulation of cyclin D3 and cyclin A . Taken together , these findings reveal a novel function of P06748 - Q9UM73 , phosphorylation and activation of JNK and cJun , which may contribute to uncontrolled cell-cycle progression and oncogenesis . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Prolonged effects of tumor necrosis factor-alpha on anterior pituitary hormone release . We examined the chronic ( 72 h ) effects of 30 ng/ml recombinant murine tumor necrosis factor ( P01375 ) -alpha on release of immunoreactive growth hormone ( GH ) , prolactin ( PRL ) , thyrotropin ( DB00024 ) , and DB00024 glycosylation , as assessed by lectin binding , in cultured rat anterior pituitary cells . In cultured cells from adult female rats , P01375 significantly suppressed basal and GH-releasing hormone ( P01148 ) -stimulated GH release . P01375 also suppressed basal PRL release and completely abolished the PRL response to TRH ( 0.1-10 nM ) . Whereas P01375 reduced basal DB00024 release , it significantly enhanced the maximal DB00024 response to TRH . P01375 did not affect the concanavalin A and lentil lectin binding of DB00024 accumulated in the medium during the 4-day culture , but significantly decreased the lentil lectin binding of DB00024 released in response to acute TRH stimulation . P01375 significantly enhanced the inhibitory effect of somatostatin on stimulated PRL release , but not on GH or DB00024 release . Compared to cell cultures from adult female rats , in anterior pituitary cell cultures from 12-day-old rats the effects of prolonged exposure to P01375 on hormone release were diminished or absent . Pituitary hormone release was unaffected by acute ( 3 h ) exposure to P01375 . These results demonstrate a direct effect of P01375 on anterior pituitary hormone release , which is cell-type specific and age dependent . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . DB08870 followed by allogeneic transplantation as salvage regimen in patients with relapsed and/or refractory Hodgkin 's lymphoma . Patients with relapsed or refractory Hodgkin lymphoma ( RR-HL ) have poor outcomes . DB08870 ( BV ) , an antibody-drug conjugate comprising an anti- P28908 antibody conjugated to the potent anti-microtubule agent , monomethyl auristatin E , induces high tumour responses with moderate adverse effects . In a retrospective study , we describe objective response rates and subsequent allogeneic stem cell transplantation ( allo- P09683 ) in patients with RR-HL treated by BV in a named patient program in two French institutions . Twenty-four adult patients with histologically proven P28908 (+) RR-HL treated with BV were included from July 2009 to November 2012 . Response to BV treatment was evaluated after four cycles . Eleven patients were in complete response ( 45.8 % ) , while five patients were in partial response ( 20.8 % ) , with an overall response rate of 66.6 % . Eight patients failed to respond to BV ( 33.3 % ) . All of the responding patients could receive consolidation treatment after BV : three patients underwent autologous stem cell transplantation ( auto- P09683 ) , three patients received a tandem auto- P09683 /allo- P09683 , nine patients received allo- P09683 and one patient was treated with donor lymphocyte infusion . We found no treatment-related mortality at day 100 among the 12 patients who underwent BV following by allogeneic transplantation . With a median follow-up of 20 months ( range 10.5-43.2 ) , none of them relapsed or died . BV followed by allo- P09683 represents an effective salvage regimen in patients with RR-HL . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain .

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interacts_with DB00013?

multiple_choice

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . Prognostic significance of urokinase plasminogen activator receptor and its cleaved forms in blood from patients with non-small cell lung cancer . DB00013 plasminogen activator ( uPA ) cleaves its three-domain cell surface receptor , Q03405 , liberating domain I [ Q03405 (I) ] and leaving the cleaved Q03405 (II-III) on the cell surface . Both intact and cleaved Q03405 can be shed from the cell surface . Q03405 (I) was previously shown to be a prognostic factor in lung tumour extracts . Here we analyse Q03405 forms in blood from patients with non-small cell lung cancer ( NSCLC ) . Preoperatively sampled plasma/serum from 32 patients with NSCLC was analysed . Three time-resolved fluoroimmunoassays ( TR-FIAs ) measuring intact Q03405 (I-III) ( TR-FIA 1 ) , Q03405 (I-III) + Q03405 (II-III) ( TR-FIA 2 ) and Q03405 (I) ( TR-FIA 3 ) were applied . The Spearman rank correlations between plasma and serum levels of Q03405 (I-III) , Q03405 (I-III) + Q03405 (II-III) , and Q03405 (I) were 0.89 , 0.94 and 0.68 respectively . Survival analysis demonstrated that high levels of all Q03405 forms were associated with shorter survival . Adjusted for histological subtype high plasma Q03405 (I-III) and Q03405 (I) levels as well as serum Q03405 (I) levels were significantly associated with shorter OS ( hazards ratios = 4.3 , 2.8 and 3.8 respectively ) . High blood levels of intact Q03405 and its cleaved forms are associated with poor prognosis in NSCLC . Increased P05231 levels in pituitary-deficient patients are independently related to their carotid intima-media thickness . OBJECTIVE : Increased cardiovascular morbidity and mortality has been observed in patients with pituitary deficiency and untreated growth hormone deficiency ( GHD ) . We investigated peripheral inflammatory and fibrinolytic markers and their associations with arterial intima-media thickness ( IMT ) in GHD . DESIGN : Cross-sectional study . PATIENTS : Thirty-four patients with GHD , but without cardiovascular disease , were compared to healthy controls matched for age , sex , body mass index ( BMI ) and smoking habits . MEASUREMENTS : IMT of the common carotid artery , P02741 ( CRP ) , interleukin-6 ( P05231 ) , fibrinogen , plasminogen activator inhibitor-1 ( P05121 ) activity and tissue plasminogen activator antigen ( tPA-ag ) were measured . RESULTS : Median P05231 concentrations were increased by 208 % and 248 % in GHD patients compared to BMI-matched and nonobese controls , respectively . Median CRP and tPA-ag levels were increased by 237 % and 167 % in patients compared to nonobese controls , but not significantly different compared to BMI-matched controls . Plasma levels of fibrinogen and P05121 activity did not differ between groups . Age , low-density lipoprotein ( LDL ) cholesterol , tPA-ag and P05231 were positively correlated , and P05019 was negatively correlated to IMT in the patient group , but only age and P05231 were independently related to IMT . CONCLUSIONS : P05231 concentrations were increased in GHD patients compared to controls and independently related to IMT in patients . This finding may help to explain the variance in IMT and the increased vascular morbidity and mortality in hypopituitary patients with GHD . Increased serum osteoprotegerin in patients with primary adrenal insufficiency receiving conventional hydrocortisone substitution . Patients treated for primary adrenal insufficiency ( P05121 ) are at risk of steroid over-replacement , which may affect their skeleton . The study was aimed to investigate the effect of steroid substitution on serum osteoprotegerin and receptor activator of nuclear factor kappa-beta ligand ( O14788 ) levels in relation to bone mineral density ( BMD ) in P05121 . Eighty patients ( mean age 47.2±14.5 years , mean hydrocortisone dose 0.49±0.14 mg/kg/day ) and 63 healthy subjects were included . Serum osteoprotegerin , O14788 , 25-hydroxyvitamin D₃ , calcium , phosphate , alkaline phosphatase , intact parathormone , and dehydroepiandrosterone-sulfate levels were evaluated in patients and controls . BMD was assessed in affected subjects using dual-energy X-ray absorptiometry . Mean osteoprotegerin concentration in P05121 patients appeared significantly higher vs. controls ( p=0.002 ) , while O14788 levels were similar ( p=0.430 ) . Serum osteoprotegerin increased with age ( p < 0.001 ) , but showed no correlation with daily hydrocortisone dose . O00300 was negatively correlated with serum dehydroepiandrosterone-sulfate ( p=0.008 ) and with BMD at the lumbar spine ( p < 0.001 ) and femoral neck ( p=0.003 ) . O14788 correlated negatively with P05121 duration ( p=0.029 ) and positively with daily hydrocortisone dose ( p=0.018 ) . Lumbar spine osteoporosis and osteopenia were found in 12 and 31 patients , respectively , whereas in femoral neck : in 5 and 33 individuals . Patients with osteoporosis displayed higher osteoprotegerin levels , but after the age-adjustment the correlation was lost . In conclusion , increased osteoprotegerin in P05121 might reflect a compensatory response to enhanced bone resorption due to exogenous steroid excess and/or result from a deficit in adrenal androgens . O14788 levels remain within normal range on standard steroid replacement . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Comparison of effects of clodronate and zoledronic acid on the repair of maxilla surgical wounds - histomorphometric , receptor activator of nuclear factor-kB ligand , osteoprotegerin , P04275 , and caspase-3 evaluation . BACKGROUND : The aim of this study was to compare clodronate and zoledronic acid regarding their influence on the repair of surgical wounds in maxillae ( soft tissue wound and tooth extraction ) and their relation to osteonecrosis . MATERIAL AND METHODS : Thirty-four Wistar rats were allocated into three groups according to the treatment received : ( i ) 12 animals treated with zoledronic acid , ( ii ) 12 animals treated with clodronate and ( iii ) 10 animals that were given saline solution . All animals were subjected to tooth extractions and surgically induced soft tissue injury . Histological analysis of the wound sites was performed by means of hematoxylin-eosin ( H & E ) staining and immunohistochemical staining for receptor activator of nuclear factor-kB ligand ( O14788 ) , osteoprotegerin ( O00300 ) , P04275 , and caspase-3 . RESULTS : The zoledronic acid group showed higher incidence of non-vital bone than did the clodronate group at the tooth extraction site . At the soft tissue wound site , there were no significant differences in non-vital bone between the test groups . O14788 , O00300 , P04275 , and caspase-3 did not show significant differences between the groups for both sites of surgical procedures . CONCLUSION : Both of the bisphosphonates zoledronic acid and clodronate are capable of inducing maxillary osteonecrosis . Immunohistochemical analysis suggests that the involvement of soft tissues as the initiator of osteonecrosis development is less probable than has been pointed out . Comparison of expression profiles induced by dust mite in airway epithelia reveals a common pathway . BACKGROUND : Airway epithelial cells have shown to be active participants in the defense against pathogens by producing signaling and other regulatory molecules in response to the encounter . METHODS : In previous manuscripts , we have studied the effect of house dust mite ( HDM ) extract on both an epithelial cell-line ( H292 ) and primary nasal epithelial cell . When we compare these responses we conclude that the H292 cells more closely resemble nasal epithelium of healthy controls ( share 107 probe-sets ) than of allergic individuals ( share 17 probe-sets ) . RESULTS : Interestingly , probably because of an absent intraindividual variation between samples , more probe-sets ( 8280 ) change expression significantly in H292 than in either healthy ( 555 ) or allergic ( 401 ) epithelium . CONCLUSIONS : A direct comparison of all the responses in these epithelial cells reveals a core-response to HDM of just 29 genes . These genes ( P78556 , P10145 , P19875 , P09341 , IL-1B , P15514 , P21580 , Q99075 , P35354 , P12643 , P01130 , Q03405 , P00749 , Q00653 , P19838 , P05412 , P18847 , P18146 , O15118 , Q8IUC6 , P29317 , P29279 , P28562 , O43609 , TLR-3 , complement factor P01024 , Q9Y6Y0 , SerpinB3 , and Q9Y617 ) have described links with allergy or inflammation and may even describe the well-established relationship between viral infections and allergic exacerbations or allergy development . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Expression of urokinase , plasminogen activator inhibitors and urokinase receptor in pregnant rhesus monkey uterus during early placentation . We have investigated plasmin mediated proteolysis associated with trophoblast invasion during early stages of pregnancy in the rhesus monkey . In situ hybridization and immunocytochemical localization were used to define the cellular and tissue distribution of urokinase plasminogen activator ( uPA ) , plasminogen activator inhibitor type 1 ( P05121 ) and 2 ( P05120 ) and urokinase receptor in early monkey placenta and uterus . Our results indicate : ( 1 ) uPA is expressed in proliferating and invasive cytotrophoblast located in chorionic villi as well as in extravillous trophoblast associated with uterine arterioles . This raises the possibility that urokinase may play an important role in trophoblast invasion . ( 2 ) P05121 mRNA is specifically localized in two areas where invasive trophoblast cells encounter maternal tissue directly . The extravillous cytotrophoblast cells at the maternofetal junction express P05121 mRNA . The invasive endovascular trophoblast cells within the uterine arterioles also express P05121 mRNA . The location sensitive expression of P05121 mRNA at the maternofetal junction may imply a protective function of this protease inhibitor that might be induced through interaction with decidual cells . ( 3 ) DB00013 receptor antigen has also been found at the maternofetal junction and in endovascular trophoblast cells of the invaded maternal blood vessel . ( 4 ) P05120 immunoreactivity is found in association with cytotrophoblast cells in anchoring choronic villi suggesting its association with early placentation . In conclusion , we propose that the plasmin/plasminogen activator system may not only regulate extracellular matrix degradation , but also modify migration and invasive behaviour of extravillous trophoblast cells , during early placentation . The role of the urokinase receptor in extracellular matrix degradation by HT29 human colon carcinoma cells . DB00013 ( u-PA ) and the urokinase receptor ( u-PAR ) , are thought to play a critical role in the invasive and metastatic properties of cancer cells . The HT29 human colon-carcinoma cell line was selected to evaluate these aspects . HT29 cells express u-PA receptors ( 100,000 sites/cell , KD = 1.5 nM ) , but no PA activity and therefore are unable to generate plasmin in the presence of plasminogen . These cells have been transfected with a human u-PA cDNA to investigate whether secreted u-PA would enhance in vitro extracellular matrix degradation , and whether the binding of u-PA to the cell surface is determinant . Five clones were selected for stable expression of high PA activity . These clones were capable of marked plasminogen-dependent degradation of R22 smooth-muscle-cell-derived extracellular matrix , whereas the parental cell line contributed to an insignificant breakdown only . DB06692 , polyclonal anti-u-PA IgG , recombinant P05120 , and co-culture with human P05121 -I-producing mouse L cells significantly inhibited this degradation . Furthermore , a peptide displacing u-PA from its receptor as well as 2 different polyclonal anti-u-PA receptor IgGs decreased the breakdown after 24 hr by as much as 70 % and 81 % , respectively . These results show that the binding of u-PA to its receptor plays an important role in in vitro matrix breakdown by HT29 u-PA transfectants . Uptake of particulate vaccine adjuvants by dendritic cells activates the Q96P20 inflammasome . Many currently used and candidate vaccine adjuvants are particulate in nature , but their mechanism of action is not well understood . Here , we show that particulate adjuvants , including biodegradable poly(lactide-co-glycolide) ( P00747 ) and polystyrene microparticles , dramatically enhance secretion of interleukin-1beta ( IL-1beta ) by dendritic cells ( DCs ) . The ability of particulates to promote IL-1beta secretion and caspase 1 activation required particle uptake by DCs and Q96P20 . Uptake of microparticles induced lysosomal damage , whereas particle-mediated enhancement of IL-1beta secretion required phagosomal acidification and the lysosomal cysteine protease cathepsin B , suggesting a role for lysosomal damage in inflammasome activation . Although the presence of a Toll-like receptor ( TLR ) agonist was required to induce IL-1beta production in vitro , injection of the adjuvants in the absence of TLR agonists induced IL-1beta production at the injection site , indicating that endogenous factors can synergize with particulates to promote inflammasome activation . The enhancement of antigen-specific antibody production by P00747 microparticles was independent of Q96P20 . However , the ability of P00747 microparticles to promote antigen-specific P05231 production by T cells and the recruitment and activation of a population of CD11b(+)Gr1(-) cells required Q96P20 . Our data demonstrate that uptake of microparticulate adjuvants by DCs activates the Q96P20 inflammasome , and this contributes to their enhancing effects on innate and antigen-specific cellular immunity . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . P04626 mRNA status contributes to the discrepancy between gene amplification and protein overexpression in gastric cancer . BACKGROUND : Human epidermal growth factor receptor 2 ( P04626 ) is an important proto-oncogene of prognostic use in gastric cancer ( GC ) . Fluorescence in-situ hybridization ( Q5TCZ1 ) and immunohistochemistry ( IHC ) are the main clinical methods of detection of P04626 , but consistency between the methods is poor and the cause of the discrepancy is unclear . AIM : To investigate the involvement of P04626 mRNA status in the disparity between gene amplification and protein overexpression . METHODS : We investigated P04626 gene , mRNA , and protein profiles in gastric precancer and cancer tissues by use of the molecular approaches Q5TCZ1 , real-time polymerase chain reaction , and IHC . The relationships between P04626 and matrix metalloproteinase 9 ( P14780 ) and O15105 expression were analyzed and the involvement of P04626 in the interaction between tumor cells and lymphocytes was investigated by coculturing GC cell lines with peripheral blood mononuclear cells ( PBMCs ) . RESULTS : P04626 protein expression was significantly increased in cancer compared with precancer ( P = 0.003 ) , and the corresponding mRNA levels were significantly lower in precancer and cancer tissues than in normal tissues ( κ = 0.290 , P = 0.025 ) . P04626 mRNA levels were significantly higher in tumor than in peritumor tissue ( P = 0.028 ) , and were positively correlated with P14780 and O15105 mRNA levels in tumor tissues . P04626 mRNA expression in GC cell lines was increased by coculture with PBMCs . CONCLUSIONS : Different P04626 mRNA profiles , possibly in relation to contact between tumor cells and lymphocytes , might help to explain the discrepancy between gene amplification and protein overexpression results . The plasminogen activator urokinase and its inhibitor P05120 in endometrial cancer . Invasion and metastasis of malignant cells require the disruption of the extracellular matrix , degradation of basement membranes , and intrusion into connective tissue and vascular and lymphatic spaces . Several studies have indicated a role for urokinase ( u-PA ) in proteolysis of the extracellular matrix and hence in stromal invasion and metastasis . Many malignant cells are known to secrete u-PA . P00747 activator inhibitor-type 2 ( P05120 ) is an inhibitor of u-PA and is present in several neoplastic cell lines and malignant ascites . We measured u-PA and P05120 antigen in tissue homogenates of normal and malignant endometrium from 21 postmenopausal patients . Enzyme-linked immunoassays which measure the bound and unbound , single-and two-chain form of the activator and bound and unbound form of the inhibitor were used . DB00013 was present in four of seven normal ( range , 0.15-0.5 ; median , 0.15 ng/mg protein ) and in significantly higher concentrations in all malignant endometrial homogenates ( range , 0.41-9.2 ; median , 3.4 ng/mg protein ) , P < 0.001 . P05120 was detectable in four of seven normal endometrial homogenates at low concentrations ( range , 1.1-3.1 ; median , 1.1 ng/mg protein ) and in all malignant tissue homogenates at significantly higher levels ( range , 1.6-27.3 ; median , 4.9 ng/mg protein ) , P < 0.01 . Levels of endometrial P05120 were higher in stages IC or greater compared to those in stages IA and 1B cancers ( P < 0.05 ) . P05120 may be useful as a prognostic marker in endometrial cancer . Increased levels of Candida albicans mannan-specific T-cell-derived antigen binding molecules in patients with invasive candidiasis . In addition to cytokines , P01730 + T cells have been found to secrete soluble , T-cell-derived antigen binding molecules ( TABMs ) . These antigen-specific immunoproteins are thought to have immunoregulatory properties in the suppression of cell-mediated immunity ( CMI ) because they often associate with interleukin-10 ( P22301 ) and transforming growth factor beta . Decreased CMI causes susceptibility to infections caused by organisms which are normally nonpathogenic . In this situation , e.g. , Candida albicans saprophytism may develop into invasive candidiasis . The difficult diagnosis of invasive candidiasis is based on the findings obtained from blood cultures and with tissue biopsy specimens , with some additional diagnostic value gained by the detection of Candida albicans mannan antigenemia and antimannan antibodies . In the present study , Candida albicans mannan-specific TABM ( P62158 -TABM ) levels in the sera of patients with invasive candidiasis ( n = 11 ) , Candida colonization ( n = 11 ) and noncolonization ( n = 10 ) , recurrent vulvovaginal candidiasis ( n = 30 ) , and atopic eczema dermatitis syndrome ( n = 59 ) and healthy controls ( n = 30 ) were analyzed . For 14 participants , the effect of mannan stimulation on TABM production and gamma interferon ( P01579 ) and P05112 mRNA expression by peripheral blood lymphocytes was also studied . It was demonstrated that P62158 -TABM production was the highest in patients with invasive candidiasis and that P62158 -TABM levels could distinguish Candida-colonized patients from noncolonized patients . In addition , the P62158 -TABM level was directly related to mRNA expression for P05112 but not P01579 . These results reinforce the view that TABMs are associated with decreased CMI , immunoregulation , and the T-helper cell 2-type immune response . DB00013 plasminogen activator and its inhibitor , P05121 , as prognostic markers in breast cancer : from pilot to level 1 evidence studies . BACKGROUND : For optimum management of patients with cancer , accurate assessment of prognosis is essential . The primary determinant of outcome in malignancy is the formation of distant metastases . DB00013 plasminogen activator ( uPA ) is a serine protease causally involved in invasion and metastasis . CONTENT : Data from model systems show that uPA is unequivocally involved in cancer dissemination . Consistent with its role in metastasis , multiple independent groups have shown that high uPA concentrations in primary breast cancers correlate with poor prognosis . For determining outcome , the prognostic impact of uPA was both independent of traditionally used factors and prognostic in patients with axillary node-negative disease . Paradoxically , high concentrations of plasminogen activator inhibitor ( P05121 ) , an endogenous inhibitor of uPA , also correlate with poor prognosis in patients with breast cancer , including the subgroup with node-negative disease . The prognostic value of uPA/ P05121 in axillary node-negative breast cancer patients was recently confirmed in both a prospective randomized trial and a pooled analysis , i.e. , two different level 1 evidence ( LOE-1 ) studies . CONCLUSIONS : uPA and P05121 are among the first biological prognostic factors to have their clinical value validated using LOE-1 evidence studies . Determination of these analytes may help identify low-risk node-negative breast cancer patients for whom adjuvant chemotherapy is unnecessary . DB00013 /urokinase receptor and vitronectin/alpha(v)beta(3) integrin induce chemotaxis and cytoskeleton reorganization through different signaling pathways . P04004 ( VN ) and pro-urokinase ( pro-uPA ) stimulated migration of rat smooth muscle cells in a dose-dependent and additive way , and induced motile-type changes in cell morphology together with a complete reorganization of the actin filaments and of the microtubules . All these effects were inhibited by pertussis toxin , or by antibodies directed against the urokinase receptor ( Q03405 ) or against the VN receptor alpha(v)beta(3) suggesting that an association between the two receptors is required to mediate both signals . Investigation of the signaling pathways showed that increasing the intracellular DB02527 resulted in a selective inhibition of VN-induced cell migration . On the other hand , PD 98059 , an inhibitor of MEK , differentially inhibited the pro-uPA- but not the VN-induced cell migration . Phosphorylation and nuclear translocation of Erk by pro-uPA was directly observed . We conclude that the signaling pathways of pro-uPA and VN must be at least in part different . MiR-221/-222 differentiate prognostic groups in advanced breast cancers and influence cell invasion . BACKGROUND : MiR-221/-222 are frequently overexpressed in breast cancer and are associated with increased malignancy . The specific modification of microRNAs ( miRNAs ) expression could be a promising strategy in breast cancer therapy , leading to the suppression of tumourigenic processes in tumour cells . METHODS : MiR-221/-222 expressions were analysed in 86 breast cancer tissues by quantitative RT-PCR and tested for correlation with immunohistochemistry data and clinical follow-up . In vitro assays were conducted using human breast cancer cell lines with lentiviral overexpression of miR-221/-222 . RESULTS : In tumour tissues , miR-221/-222 were associated with the occurrence of distant metastases . In particular , high levels of miR-221 were revealed to have a high prognostic impact for the identification of significantly different groups with advanced tumours . MiR-221/-222 overexpression strongly increased cell proliferation and invasion in vitro . Following miR-221/-222 overexpression an increased Q03405 expression and cell invasion were observed . CONCLUSION : This study demonstrates a significant role for highly expressed miR-221/-222 in advanced breast cancers allowing for the identification of significantly different prognostic groups , particularly for P04626 -positive and lymph-node-positive breast cancers . Considering that miR-221/-222 are strongly involved in cell invasion , these miRNAs may be promising markers for breast cancer prognosis and therapy . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . P00747 activator inhibitor-1 and vitronectin expression level and stoichiometry regulate vascular smooth muscle cell migration through physiological collagen matrices . BACKGROUND : Vascular smooth muscle cell ( VSMC ) migration is a critical process in arterial remodeling . Purified plasminogen activator inhibitor-1 ( P05121 ) is reported to both promote and inhibit VSMC migration on two-dimensional ( D ) surfaces . OBJECTIVE : To determine the effects of P05121 and vitronectin ( VN ) expressed by VSMC themselves on migration through physiological collagen matrices . METHODS : We studied migration of wild-type ( WT ) , P05121 -deficient , VN-deficient , P05121 /VN doubly-deficient ( DKO ) and P05121 -transgenic ( Tg ) VSMC through three-D collagen gels . RESULTS : WT VSMC migrated significantly slower than P05121 - and VN-deficient VSMC , but significantly faster than DKO VSMC . Experiments with recombinant P05121 suggested that basal VSMC P05121 expression inhibits migration by binding VN , which is secreted by VSMC and binds collagen . However , P05121 -over-expressing Tg VSMC migrated faster than WT VSMC . Reconstitution experiments with recombinant P05121 mutants suggested that the pro-migratory effect of P05121 over-expression required its anti-plasminogen activator ( PA ) and P01130 -related protein ( Q14764 ) binding functions , but not VN binding . While promoting VSMC migration in the absence of P05121 , VN inhibited the pro-migratory effect of active P05121 . CONCLUSIONS : In isolation , VN and P05121 are each pro-migratory . However , via formation of a high-affinity , non-motogenic complex , P05121 and VN each buffers the other 's pro-migratory effect . The level of P05121 expression by VSMC and the concentration of VN in extracellular matrix are critical determinants of whether P05121 and VN promote or inhibit migration . These findings help to rectify previously conflicting reports and suggest that P05121 /VN stoichiometry plays an important role in VSMC migration and vascular remodeling . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . The effect of Plasmodium falciparum infection on expression of monocyte surface molecules . Plasmodium falciparum infection may result in severe malaria in susceptible individuals . The pathogenesis of severe disease is probably a combination of the sequestration of infected erythrocytes and overstimulation of the immune response . Monocytes are a key source of many of the pro-inflammatory agents implicated but also are found sequestered in blood vessels . However , little is known about the monocyte phenotype in malaria disease . Flow cytometry was performed on fresh whole blood to determine surface expression of four receptors during acute severe and non-severe malaria and again during convalescence when uninfected . Three hundred and fifty-six children with P. falciparum infection were studied and were found to show increased expression of intercellular adhesion molecule-1 ( P05362 ) , urokinase plasminogen activator receptor ( Q03405 ) , CD23 and chemokine receptor 5 ( P51681 ) ( P < 0.001 ) during acute disease compared with convalescent levels . Using multivariate analysis , it was found that large increases in expression of P05362 ( odds ratio ( OR ) 2.44 , 95 % CI 1.80-3.32 ) and Q03405 ( OR 3.14 , 95 % CI 1.93-5.09 ) but small increases in expression of CD23 ( OR 0.82 , 95 % CI 0.68-0.96 ) were independently associated with severe malaria . These results give an insight into the cellular processes occurring in severe malaria and suggest that pathology is based on a complex repertoire of pro- and anti-inflammatory processes . Elevated urokinase-specific surface receptor expression is maintained through its interaction with urokinase plasminogen activator . DB00013 plasminogen activator ( uPA ) and its receptor ( Q03405 ) are overexpressed in various neoplasms , and play a key role in tumor progression and metastasis . In this study , we examined uPA and Q03405 expression in a variety of human breast cancer cell lines and found that lines with elevated uPA expression also exhibited high Q03405 expression , suggesting the possibility that uPA and Q03405 are regulated in concert . To test this possibility , we introduced antisense uPA RNA and antisense Q03405 RNA in MDA-MB-231 and BT-549 lines that express high levels of uPA and Q03405 . Antisense uPA RNA not only downregulated uPA expression , but also greatly reduced Q03405 expression in both lines . However , antisense Q03405 RNA-reduced Q03405 expression with no apparent inhibitory effect on the levels of uPA . These results indicate that expression of Q03405 requires uPA but not vice versa . With a panel of uPA and Q03405 monoclonal antibodies ( mAbs ) , we observed that the mAbs disrupting uPA and Q03405 interaction , rather than mAb inhibiting uPA protease activity , reduced Q03405 expression . Moreover , adding soluble single chain uPA ( scuPA ) to MDA-MB-231 or BT-549 cells expressing antisense uPA mRNA-restored Q03405 expression . These findings suggest that uPA dictates Q03405 expression and that uPA binding to Q03405 transmits signals for Q03405 expression . Finally , we provided evidence that Fyn , a Src family kinase , is involved in uPA-induced Q03405 expression . Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . DB00013 plasminogen activator stimulates function of active forms of stromelysin and gelatinases ( P08253 and P14780 ) in cirrhotic tissue . BACKGROUND : The authors ' previous data support the notion that adenoviral-driven urokinase plasminogen activator ( u-PA ) expression results in reversion of experimental liver cirrhosis . The specific aim of the present study was to decipher the mechanisms involved in the regulation by endogenous/gene-delivered u-PA of matrix metalloproteinases ( MMP ) and related proteins engaged in degradation of excessive hepatic connective tissue . METHODS : Tissue slices from cirrhotic rat livers were incubated with u-PA-rich supernatants from 24-h-cultured hepatic stellate cells ( P19526 ) . P08253 , -9 and tissue inhibitor of metalloproteinases-1 ( P01033 ) were detected by western blot and biologic activity . The P19526 that discontinued u-PA production were transfected with the adenovector Adu-PA and serum-free supernatants evaluated for proteolytic activity by P08254 , P08253 and P14780 . Collagen I , transforming growth factor-beta1 ( TGF-beta1 ) , plasminogen activator inhibitor-1 ( P05121 ) and P01033 mRNA levels were also evaluated . RESULTS AND CONCLUSION : Endogenous u-PA from cultured P19526 significantly induced the active forms of P08253 ( 68 kDa ) and P14780 ( 78 kDa ) in cirrhotic tissue slices . The P01033 molecular forms demonstrated that u-PA pushed the presence of ' free ' P01033 ( not complexed with MMP ; 71 % ) in cirrhotic tissue . When non-producing u-PA- P19526 were transfected with adenoviral vector coding for the functional human protein u-PA ( Adhu-PA ) , an overactivation of P08254 , P08253 and P14780 ( 800 % , 48 % and 100 % , respectively ) was found as compared with P19526 transfected with control adenovirus encoding green fluorescent protein ( Ad-GFP ) . Finally , gene expression of collagen I , TGF-beta1 , P05121 and P01033 were downregulated by Adhu-PA action as well . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . Innate immune responses to Q9NP99 activation : overlap , divergence , and positive and negative cross-talk with bacterial lipopolysaccharide . Q9NP99 ( triggering receptor expressed on myeloid cells-1 ) is an orphan immunoreceptor expressed on monocytes , macrophages , and neutrophils . Q9NP99 associates with and signals via the adapter protein O43914 / O43914 , which contains an ITAM . Q9NP99 activation by receptor cross-linking has been shown to be proinflammatory and to amplify some cellular responses to TLR ligands such as bacterial LPS . To investigate the cellular consequences of Q9NP99 activation , we have characterized global gene expression changes in human monocytes in response to Q9NP99 cross-linking in comparison to and combined with LPS . Both Q9NP99 activation and LPS up-regulate chemokines , cytokines , matrix metalloproteases , and PTGS/ P35354 , consistent with a core inflammatory response . However , other immunomodulatory factors are selectively induced , including P10451 and P09603 ( i.e. , P09603 ) by Q9NP99 activation and IL-23 and P09919 ( i.e. , DB00099 ) by LPS . Additionally , cross-talk between Q9NP99 activation and LPS occurs on multiple levels . Although synergy in GM- P04141 protein production is reflected in commensurate mRNA abundance , comparable synergy in IL-1beta protein production is not . Q9NP99 activation also attenuates the induction of some LPS target genes , including those that encode IL-12 cytokine family subunits . Where tested , positive Q9NP99 outputs are greatly reduced by the PI3K inhibitor wortmannin , whereas this attenuation is largely PI3K independent . These experiments provide a detailed analysis of the cellular consequences of Q9NP99 activation and highlight the complexity in signal integration between ITAM- and TLR-mediated signaling . Insights in the molecular mechanisms of the anti-angiogenic effect of an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase . 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors ( statins ) reduce the risk of coronary event by cholesterollowering dependent and independent mechanisms . We have already described that the inhibitory effect of cerivastatin on angiogenesis contribute to the cholesterol-independent beneficial effect and was due to the inhibition of the cell signaling cascade RhoA/ Q05397 /Akt . In this study , new insights in the molecular mechanism of action were provided . It indicates an inhibition of exposure of alpha V beta 3 integrin on cell membrane and a modification of gene expression . The inhibition of angiogenesis could be related to 1 ) an increase in genes involved in the inhibition of cell proliferation ( p19(INK4) , P38936 (Waf/Cip1),Wnt-5a ) , the inhibition of cell migration ( Rho-GDI 1 , alpha E-catenin ) and 2 ) a downregulation of genes involved in angiogenesis ( P05121 , P04004 , HoxD3 , Notch4 ) or in cell invasion ( Semaphorin E ) . In addition , DNA repair protein genes ( P40692 , P18887 ) were increased . This study may indicate new biological interest of genes involved in angiogenesis control . Restoration of flow following haemodialysis catheter thrombus . Analysis of rt-PA infusion in tunnelled dialysis catheters . PROBLEM : DB00013 and streptokinase are commonly used thrombolytic agents for obstructed central venous catheters . Although proven to be efficacious , these agents have the potential to induce fibrin breakdown and streptokinase can not be used repeatedly because of its allergenic nature . Published evidence suggests that DB00013 is safe and effective ( > 70 % efficacy for catheter installation ) and that P00750 ( rt-PA ) can achieve as much as 98 % success . OBJECTIVE : To describe our experience with and our protocol for the use of rt-PA as an alternative agent for catheter thrombolysis . DESIGN : Investigation of a cohort of haemodialysis patients with tunnelled central venous catheter ( SPCVC ) placed between December 2001 to August 2003 and who developed catheter thrombus ( female , n = 8 : male , n = 12 ) . Each patient was given an infusion of between 1 and 2 mg rt-PA/h for 4 h . The dose was dependent on partial or total line obstruction . The technical success of rt-PA is defined as returning catheter blood flow to > 250 mL/min for a 4-h period . FINDINGS : Twenty patients required 57 infusions in 38 lumens between 01/01/02 to 01/09/03 . For completely blocked lines rt-PA was infused at 2 mg/h for 4 h achieving 85 % success rate . For inadequate flow ( < 250 mL/min ) rt-PA was infused at 1 mg/h for 4 h achieving an 88 % success rate . CONCLUSION : Rt-PA administered at 2 mg/h for blocked lines effectively restores haemodialysis catheter patency , and at 1 mg/h for sluggish lines is also effective in restoring blood flow through catheters . DB00013 plasminogen activator receptor ( Q03405 ) and plasminogen activator inhibitor-1 ( P05121 ) are potential predictive biomarkers in early stage oral squamous cell carcinomas ( OSCC ) . Oral squamous cell carcinoma ( OSCC ) is often associated with metastatic disease and a poor 5 year survival rate . Patients diagnosed with small tumours generally have a more favourable outcome , but some of these small tumours are aggressive and lead to early death . To avoid harmful overtreatment of patients with favourable prognosis , there is a need for predictive biomarkers that can be used for treatment stratification . In this study we assessed the possibility to use components of the plasminogen activator ( PA ) system as prognostic markers for OSCC outcome and compared this to the commonly used biomarker Ki-67 . A tissue-micro-array ( TMA ) based immunohistochemical analysis of primary tumour tissue obtained from a North Norwegian cohort of 115 patients diagnosed with OSCC was conducted . The expression of the biomarkers was compared with clinicopathological variables and disease specific death . The statistical analyses revealed that low expression of Q03405 ( p = 0.031 ) and P05121 ( p = 0.021 ) in the tumour cells was significantly associated with low disease specific death in patients with small tumours and no lymph node metastasis ( T1N0 ) . The commonly used biomarker , Ki-67 , was not associated with disease specific death in any of the groups of patients analysed . The conclusion is that Q03405 and P05121 are potential predictive biomarkers in early stage tumours and that this warrants further studies on a larger cohort of patients . DB00013 plasminogen activator ( uPA ) and its receptor ( Q03405 ) in gestational tissues ; Measurements and clinical implications . BACKGROUND : DB00013 plasminogen activator ( uPA ) and urokinase plasminogen activator receptor ( Q03405 ) are central molecules for uPA/ Q03405 /plasmin-dependent proteolysis , which is thought to play a significant role in the development of pregnancy , as well as its many complications . OBJECTIVE : To measure the levels of uPA and Q03405 in the placenta and myometrium , as well as in the foetal membranes and amniotic fluid . STUDY DESIGN : The study group consisted of 35 women with normal course of pregnancy , but with complications arising during delivery , which led to Caesarean section . Samples of placenta , myometrium , foetal membranes , amniotic fluid and blood were obtained at the time of operation . Tissue extracts were prepared . Measurements were made by the ELISA method . RESULTS : uPA and Q03405 concentration in gestational tissues , including amniotic fluid , is 100-200 times higher than in plasma . Among tissues , the highest uPA level was found in placenta ( 1.32 +/- 0.48 ng/mg of protein ) , and the highest Q03405 level in foetal membranes ( 3.33 +/- 1.20 ng/mg of protein ) . CONCLUSIONS : uPA and Q03405 are present in all gestational tissues , in some in relatively high concentrations . Our results support the modern clinical hypothesis that fibrinolytic system can participate in mechanisms of such obstetric complications as pre-term pre-mature rupture of foetal membranes and placental abruption . DB00013 plasminogen activator receptor promotes macrophage infiltration into the vascular wall of ApoE deficient mice . The urokinase plasminogen activator receptor ( Q03405 ) regulates macrophage adhesion and migration by binding directly to matrix proteins and signaling through integrin complexes . In this study , we examined the role of Q03405 on macrophage infiltration into the vascular wall . Stable murine macrophage ( Raw264.7 ) cell lines expressing high levels of human Q03405 , human urokinase plasminogen activator ( uPA ) , or both were established using expression vectors driven by the human P34810 promoter . Stimulation with human uPA specifically induced phosphorylation of early response regulated kinase ( P29323 ) in cells expressing human Q03405 but not in sham transfected cells . The human Q03405 expressing Raw264.7 cells showed increased adhesion to both human uPA and vitronectin ( Vn ) . Raw264.7 cells expressing human Q03405 or both human Q03405 and uPA , but not uPA alone , were detected in the aortic wall of ApoE(-/-) mice , and no cells were detected in that of age-matched C57BL/6J mice after intravenous infusion of the cells . Blocking of Mac-1/ P05362 interaction by anti-alphaM antibody ( M1/70 ) significantly reduced the infiltration of huPAR-expressing Raw264.1 cells into aorta of ApoE(-/-) mice . Treatment of C57BL/6J mice with angiotensin II resulted in infiltration of Raw264.7 cells expressing human Q03405 . These data demonstrate that Q03405 plays a key role in promoting macrophage infiltration into the arterial wall of ApoE(-/-) mice . Knock-down of calcitonin receptor expression induces apoptosis and growth arrest of prostate cancer cells . P01258 ( CT ) and its receptor ( CTR ) are expressed only in basal epithelium of benign prostate and in whole epithelium of malignant prostates . Also , CT and CTR mRNA levels in prostate cancers increase with an increase in tumor grade . We tested the role of the CT/CTR autocrine axis on the tumorigenicity of prostate cancer cells . We enforced the expression of CTR in CT-positive/CTR-deficient PC-3 cells . In contrast , we knocked down CTR expression in CT/CTR-positive PC-3M cells . The effect of CTR modulation on the oncogenicity was evaluated by the rate of cell proliferation , invasion , colony formation and in vivo growth in nude mice . Up-regulation of CTR in PC-3 cells and its down-regulation in PC-3M cells significantly altered their tumorigenicity . Intratumorally administered CTR RNAi in preexisting PC-3M xenografts markedly attenuated their further growth . This treatment also led to a remarkable decrease in endothelial cell populations in the tumors and increase in apoptotic , P12004 -negative cell populations . Tumors receiving CTR RNAi treatment displayed markedly lower levels of urokinase-type plasminogen activator , phospho-Akt and survivin , suggesting CTR activates uPA- Q03405 axis and P19957 -kinase-Akt-survivin pathway . These results suggest an important role for CT-CTR autocrine axis in the progression of localized prostate tumor to a metastatic phenotype , and offer a potential therapeutic option for invasive cancers . Differential expression of the urokinase receptor in fibroblasts from normal and fibrotic human lungs . Binding of urokinase-type plasminogen activator ( uPA ) to a specific receptor ( Q03405 ) on human lung fibroblasts enables it to regulate cellular proteolysis and remodeling of the extracellular matrix . Binding studies with radiolabeled uPA indicated that both normal and fibrotic lung fibroblasts express the receptor , but cells from fibrotic tissues bound significantly more uPA ( P < 0.001 ) . Phorbol myristate acetate , lipopolysaccharide , transforming growth factor-beta ( TGF-beta ) , and tumor necrosis factor-alpha ( P01375 ) increased uPA binding and plasminogen activation at the cell surface , with a greater maximal effect on fibrotic than on normal fibroblasts . Excess unlabeled uPA , specific antibody , or antisense oligonucleotides inhibited uPA binding . Ribonuclease ( RNase ) protection assays showed higher levels of Q03405 messenger ribonuleic acid ( mRNA ) in each of the five fibrotic cell lines than in normal fibroblasts . uPA was mitogenic for normal as well as fibrotic fibroblasts , indicating that receptor binding concurrently localizes cellular proteolytic activity and stimulates mitogenesis . Morphometry and immunohistochemical analysis showed that Q03405 , as well as uPA , was increased in fibroblasts in fibrotic lung tissue . Increased expression of Q03405 by fibrotic lung fibroblasts and enhanced urokinase binding induced by proinflammatory cytokines suggest a novel mechanism by which fibroblast-mediated matrix remodeling and proliferation may be regulated in interstitial lung diseases . P07996 and transforming growth factor beta-1 upregulate plasminogen activator inhibitor type 1 in pancreatic cancer . Controlled degradation of the extracellular matrix by proteases is crucial in tumor cell invasion . We have shown that thrombospondin-1 ( P07996 -1 ) , through activation of transforming growth factor beta-1 ( TGF-beta1 ) , regulates the plasminogen/plasmin protease system in breast cancer . To determine whether this occurred in other epithelial neoplasms , we studied the role of P07996 -1 and TGF-beta1 in the regulation of the plasminogen/plasmin system in pancreatic cancer . ASPC-1 and COLO-357 pancreatic cancer cells were treated with P07996 -1 or TGF-beta1 at varying concentrations . The P07996 -1 and TGF-beta1-treated cells were also treated with either anti- P07996 -1 , anti- P07996 -1 receptor , or anti-TGF-beta1 antibodies . DB00013 plasminogen activator ( uPA ) and plasminogen activator inhibitor-1 ( P05121 ) expression was determined by enzyme-linked immunosorbent assay . P07996 -1 and TGF-beta1 promoted a dose-dependent upregulation of ASPC-1 and COLO-357 P05121 expression . The P07996 -1 effect could be blocked with anti- P07996 -1 or anti-TGF-beta1 antibodies . The TGF-beta1 effect could be blocked only with anti-TGF-beta1 antibody . Anti- P07996 -1 receptor antibody blocked the P07996 -1 effect on P05121 expression but had no effect on TGF-beta1-mediated P05121 expression . Neither P07996 -1 nor TGF-beta1 had an effect on uPA production . We conclude that P07996 -1 , in a receptor-mediated process that involves the activation of TGF-beta1 , upregulates P05121 expression in pancreatic cancer without an effect on uPA production . DB00013 receptor is up-regulated in endothelial cells and macrophages associated with fibrinoid deposits in the human placenta . Clearance of fibrin deposits within the human placenta is an ongoing process during normal placental development . P00747 is a circulating fibrinolytic protease zymogen activated in situ by plasminogen activators . We have previously reported that the receptor for urokinase plasminogen activator ( Q03405 ) is expressed by cells either covering or enmeshed within the perivillous fibrinoid deposits . Whereas these cells seemed likely to be trophoblasts , a definitive identification was lacking , and this question is central to the understanding of the cellular mechanisms directing fibrinolysis in the placenta . In this study we have performed immunohistochemical co-localization studies and found that the Q03405 -positive cells covering fibrinoid deposits are immunoreactive for CD31 and P04275 , indicating that they are actually endothelial cells . In addition , we found that perivillous fibrinoid deposits not covered with Q03405 -positive endothelial cells were covered with platelets identified by integrin alpha(IIb)beta(3)-immunoreactivity . Also surprisingly , the Q03405 -positive cells enmeshed within fibrinoid deposits express a cell specific marker indicating that they are macrophages . Both Q03405 -positive cell populations also express uPA immunoreactivity . Taken together , the data suggest that both fibrinoid-covering endothelial cells and fibrinoid-enmeshed macrophages can participate in the clearance process of perivillous fibrinoid deposits formed in the human placenta . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . Ang II-stimulated migration of vascular smooth muscle cells is dependent on Q92673 in mice . Medial-to-intimal migration of SMCs is critical to atherosclerotic plaque formation and remodeling of injured arteries . Considerable amounts of the shed soluble form of the P01130 relative Q92673 ( sLR11 ) produced by intimal SMCs enhance SMC migration in vitro via upregulation of urokinase-type plasminogen activator receptor ( Q03405 ) expression . Here , we show that circulating sLR11 is a novel marker of carotid intima-media thickness ( IMT ) and that targeted disruption of the Q92673 gene greatly reduces intimal thickening of arteries through attenuation of Ang II-induced migration of SMCs . Serum concentrations of sLR11 were positively correlated with IMT in dyslipidemic subjects , and multivariable regression analysis suggested sLR11 levels as an index of IMT , independent of classical atherosclerosis risk factors . In Lr11-/- mice , femoral artery intimal thickness after cuff placement was decreased , and Ang II-stimulated migration and attachment of SMCs from these mice were largely abolished . In isolated murine SMCs , sLR11 caused membrane ruffle formation via activation of focal adhesion kinase/ P29323 /Rac1 accompanied by complex formation between Q03405 and integrin alphavbeta3 , a process accelerated by Ang II . Overproduction of sLR11 decreased the sensitivity of Ang II-induced activation pathways to inhibition by an Ang II type 1 receptor blocker in mice . Thus , we demonstrate a requirement for sLR11 in Ang II-induced SMC migration and propose what we believe is a novel role for sLR11 as a biomarker of carotid IMT . DB00072 has preferential activity against breast cancers driven by P04626 homodimers . In breast cancer cells with P04626 gene amplification , P04626 receptors exist on the cell surface as monomers , homodimers , and heterodimers with P00533 / P21860 . The therapeutic antibody trastuzumab , an approved therapy for P04626 (+) breast cancer , can not block ligand-induced P04626 heterodimers , suggesting it can not effectively inhibit P04626 signaling . Hence , P04626 oligomeric states may predict the odds of a clinical response to trastuzumab in P04626 -driven tumors . To test this hypothesis , we generated nontransformed human MCF10A mammary epithelial cells stably expressing a chimeric P04626 -FKBP molecule that could be conditionally induced to homodimerize by adding the FKBP ligand AP1510 , or instead induced to heterodimerize with P00533 or P21860 by adding the heterodimer ligands P01133 /TGFα or heregulin . AP1510 , P01133 , and heregulin each induced growth of MCF10A cells expressing P04626 -FKBP . DB00072 inhibited homodimer-mediated but not heterodimer-mediated cell growth . In contrast , the P04626 antibody pertuzumab , which blocks P04626 heterodimerization , inhibited growth induced by heregulin but not AP1510 . Lastly , the P04626 / P00533 tyrosine kinase inhibitor lapatinib blocked both homodimer- and heterodimer-induced growth . AP1510 triggered phosphorylation of Erk1/2 but not AKT , whereas trastuzumab inhibited AP1510-induced Erk1/2 phosphorylation and Shc- P04626 homodimer binding , but not TGFα-induced AKT phosphorylation . Consistent with these observations , high levels of P04626 homodimers correlated with longer time to progression following trastuzumab therapy in a cohort of patients with P04626 -overexpressing breast cancer . Together , our findings confirm the notion that P04626 oligomeric states regulate P04626 signaling , also arguing that trastuzumab sensitivity of homodimers may reflect their inability to activate the PI3K ( phosphoinositide 3-kinase ) /AKT pathway . A clinical implication of our results is that high levels of P04626 homodimers may predict a positive response to trastuzumab . [ Treatment of renal vein thromboses in the newborn ] . Surgical thrombectomy is not a rational approach to neonatal renal vein thrombosis since the occlusion mainly involves intrarenal branches rather than the main renal vein , which is even patent in some instances . Conservative management combines supportive therapy for renal failure and systemic hypertension , if needed , and either heparin or thrombolytic agents . DB00086 has proven difficult to handle in neonates and should not be used . DB00013 has been used in 18 patients but results are difficult to interpret because these cases occurred over an 18-year period . P00747 tissue activator , the latest thrombolytic agent developed , has been used in few pediatric patients . An international task force is currently studying whether or not a randomized study is warranted to provide data for standardizing thrombolytic therapy in pediatric renal vein thrombosis . A novel role for farnesyl pyrophosphate synthase in fibroblast growth factor-mediated signal transduction . P14324 ( FPPS ) catalyses the formation of a key cellular intermediate in isoprenoid metabolic pathways . Here we describe a novel role for this enzyme in fibroblast growth factor ( FGF ) -mediated signalling . We demonstrate the binding of FPPS to FGF receptors ( FGFRs ) using the yeast two-hybrid assay , pull-down assays and co-immunoprecipitation . The interaction between FPPS and FGFR is regulated by the cellular metabolic state and by treatment with P09038 . Overexpression of FPPS inhibits P09038 -induced cell proliferation , accompanied by a failure of the P09038 -mediated induction of cyclins D1 and E. Overexpression of FPPS in fibroblasts also promotes increased farnesylation of Ras , and temporally extends P09038 -stimulated activation of the Ras/ P29323 ( extracellular-signal-regulated kinase ) cascade . These data suggest that , in addition to its role in isoprenoid biosynthesis , FPPS may function as a modulator of the cellular response to FGF treatment . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Transforming growth factor-beta is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts . We have studied the mechanism of a transforming growth factor-beta ( TGF-beta ) -stimulated production of type-1 plasminogen activator inhibitor ( P05121 ) in WI-38 human lung fibroblasts . TGF-beta causes an early increase in the P05121 mRNA level which reaches a maximal 50-fold enhancement after 8 h . Blocking of protein synthesis with cycloheximide causes an equally strong increase in the level of P05121 mRNA . Quantitative studies of the effect of TGF-beta on P05121 protein levels in cell extracts and culture media by using monoclonal antibodies are consistent with the effect on P05121 mRNA . The results suggest a primary effect of TGF-beta on P05121 gene transcription , and also suggest the possibility that the transcription of this gene in non-induced cells may be suppressed by a short-lived negatively regulating protein . DB00013 -type ( u-PA ) and tissue-type ( t-PA ) plasminogen activators are decreased in the culture media of TGF-beta-treated cells concomitantly with the increase in P05121 accumulation . These findings show that a primary and important biological effect of TGF-beta may be an overall decreased extracellular proteolytic activity , and give an insight into the molecular mechanisms underlying TGF-beta action at the genetic level . DB06643 -- an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 ) , a cytokine member of the P01375 family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis . DB00013 plasminogen receptor and the fibrinolytic complex play a role in nerve repair after nerve crush in mice , and in human neuropathies . Remodeling of extracellular matrix ( Q13201 ) is a critical step in peripheral nerve regeneration . In fact , in human neuropathies , endoneurial Q13201 enriched in fibrin and vitronectin associates with poor regeneration and worse clinical prognosis . Accordingly in animal models , modification of the fibrinolytic complex activity has profound effects on nerve regeneration : high fibrinolytic activity and low levels of fibrin correlate with better nerve regeneration . The urokinase plasminogen receptor ( Q03405 ) is a major component of the fibrinolytic complex , and binding to urokinase plasminogen activator ( uPA ) promotes fibrinolysis and cell movement . Q03405 is expressed in peripheral nerves , however , little is known on its potential function on nerve development and regeneration . Thus , we investigated Q03405 null mice and observed that Q03405 is dispensable for nerve development , whereas , loss of Q03405 affects nerve regeneration . Q03405 null mice showed reduced nerve repair after sciatic nerve crush . This was a consequence of reduced fibrinolytic activity and increased deposition of endoneurial fibrin and vitronectin . Exogenous fibrinolysis in Q03405 null mice rescued nerve repair after sciatic nerve crush . Finally , we measured the fibrinolytic activity in sural nerve biopsies from patients with peripheral neuropathies . We showed that neuropathies with defective regeneration had reduced fibrinolytic activity . On the contrary , neuropathies with signs of active regeneration displayed higher fibrinolytic activity . Overall , our results suggest that enforced fibrinolysis may facilitate regeneration and outcome of peripheral neuropathies . DB00013 -catalysed cleavage of the urokinase receptor requires an intact glycolipid anchor . DB00013 ( uPA ) has the striking ability to cleave its receptor , Q03405 , thereby inactivating the binding potential of this molecule . Here we demonstrate that the glycosylphosphatidylinositol ( P06744 ) anchor of Q03405 , which is attached to the third domain , is an important determinant in governing this reaction , even though the actual cleavage occurs between the first and second domains . Purified full-length P06744 -anchored Q03405 ( P06744 - Q03405 ) proved much more susceptible to uPA-mediated cleavage than recombinant truncated soluble Q03405 ( suPAR ) , which lacks the glycolipid anchor . This was not a general difference in proteolytic susceptibility since P06744 - Q03405 and suPAR were cleaved with equal efficiency by plasmin . Since the amino acid sequences of P06744 - Q03405 and suPAR are identical except for the C-terminal truncation , the different cleavage patterns suggest that the two Q03405 variants differ in the conformation or the flexibility of the linker region between domains 1 and 2 . This was supported by the fact that an antibody to the peptide AVTYSRSRYLE , amino acids 84-94 in the linker region , recognizes P06744 - Q03405 but not suPAR . This difference in the linker region is thus caused by a difference in a remote hydrophobic region . In accordance with this model , when the hydrophobic lipid moiety was removed from the glycolipid anchor by phospholipase C , low concentrations of uPA could no longer cleave the modified P06744 - Q03405 and the reactivity to the peptide antibody was greatly decreased . Naturally occurring suPAR , purified from plasma , was found to have a similar resistance to uPA cleavage as phospholipase C-treated P06744 - Q03405 and recombinant suPAR . Immunization strategies to augment oral vaccination with DNA and viral vectors expressing HIV envelope glycoprotein . Induction of mucosal immunity to the human immunodeficiency virus ( HIV ) envelope ( env ; gp160 ) glycoprotein has been demonstrated with orally administered recombinant vaccinia virus ( rVV ) vectors and poly(DL-lactide-co-glycolide) ( P00747 ) -encapsulated plasmid DNA expressing gp160 . In this study , we investigated the effect of an oral DNA-prime/rVV-boost vaccine regimen in conjunction with adjuvants on the level of gp160-specific cellular and humoral responses in BALB/c mice . We demonstrated that DNA priming followed by a booster with rVV expressing gp160 ( vPE16 ) significantly augmented env-specific immunity in systemic and mucosal tissues of the immunized mice . Association of vPE16 with liposomes and coadministration of liposome-associated beta-glucan lentinan or P60568 /Ig-encoded plasmid DNA-encapsulated in P00747 microparticles triggered the optimal cell-mediated immune ( CMI ) responses . Lentinan was found to increase env-specific type 1 cytokine production and cytotoxic T-lymphocyte ( CTL ) activities but had no effect on humoral responses . On the other hand , P60568 /Ig-mediated increases in both type 1 and 2 activities were associated with higher levels of env-specific CTL and antibody responses . Results of these studies demonstrated the effectiveness of oral vaccines with DNA and rVV vectors in conjunction with immunomodulators in inducing specific immune responses in systemic and mucosal tissues . Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus ( FK506 ) . Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes ( EBV-CTLs ) to solid organ transplant ( SOT ) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders ( PTLDs ) . SOT recipients , however , require the continuous administration of immunosuppressive drugs to prevent graft rejection , and these agents may significantly limit the long-term persistence of transferred EBV-CTLs , precluding their use as prophylaxis . DB00864 ( FK506 ) is one of the most widely used immunosuppressive agents in SOT recipients , and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein ( P62942 ) . We have knocked down the expression of P62942 in EBV-CTLs using a specific small interfering RNA ( siRNA ) stably expressed from a retroviral vector and found that P62942 -silenced EBV-CTLs are FK506 resistant . These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity . We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model , suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Anaplastic oligodendrogliomas with 1p19q codeletion have a proneural gene expression profile . BACKGROUND : In high grade gliomas , 1p19q codeletion and P00533 amplification are mutually exclusive and predictive of dramatically different outcomes . We performed a microarray gene expression study of four high grade gliomas with 1p19q codeletion and nine with P00533 amplification , identified by CGH-array . RESULTS : The two groups of gliomas exhibited very different gene expression profiles and were consistently distinguished by unsupervised clustering analysis . One of the most striking differences was the expression of normal brain genes by oligodendrogliomas with 1p19q codeletion . These gliomas harbored a gene expression profile that partially resembled the gene expression of normal brain samples , whereas gliomas with P00533 amplification expressed many genes in common with glioblastoma cancer stem cells . The differences between the two types of gliomas and the expression of neuronal genes in gliomas with 1p19q codeletion were both validated in an independent series of 16 gliomas using real-time RT-PCR with a set of 22 genes differentiating the two groups of gliomas ( P42330 , Q96SQ7 , P12643 , Q9BQL6 , P14635 , P24941 , P36222 , Q8WZ74 , O43602 , P00533 , Q8IUC8 , P32455 , P18065 , P46940 , L1CAM , P13591 , Q13253 , Q13516 , Q86YL7 , P00750 , Q15063 , Q8IUD6 ) . Immunohistochemical study of the most differentially expressed neuronal gene , alpha-internexin , clearly differentiated the two groups of gliomas , with 1p19q codeletion gliomas showing specific staining in tumor cells . CONCLUSION : These findings provide evidence for neuronal differentiation in oligodendrogliomas with 1p19q codeletion and support the hypothesis that the cell of origin for gliomas with 1p19q codeletion could be a bi-potential progenitor cell , able to give rise to both neurons and oligodendrocytes . [ DB00013 therapy of deep vein thrombosis ( author 's transl ) ] . 30 patients with deep vein thrombosis were treated with a combination of urokinase and heparin . Clinically relevant improvement was achieved in 2/3 of them with appr . 40,000 IU/h ( 1,000,000 IU/d ) urokinase administered over a period of several days . This indicates that urokinase at this dosage offers a valuable alternative or supplementation to fibrinolytic therapy with streptokinase . With the dosage employed , routine blood coagulation tests are only minimally affected , although a strong enhancement of fibrinolytic activity can be demonstrated by the euglobulin clot lysis time . P00747 depletion - as is usually observed with streptokinase therapy - does not occur . DB00013 is well tolerated and there is only a very moderate bleeding tendency . The cost per day of urokinase therapy at the dosage employed is approximately twice that of customary streptokinase therapy . [ Genetic differentiation of residents of Central Asia from autosomal marker data ] . The gene pool of five ethnic groups of the Central Asian population was characterized using nine human-specific polymorphic insertion/deletion loci ( P12821 , P00750 , P02647 , PV92 , P05160 , A25 , B65 , P01730 , Mt-Nuc ) . It has been shown for the first time that at the P01730 locus , the frequency of Alu(-) is inversely related to the Mongoloid component of the population . For the Central Asian populations , the lowest and highest frequencies of the Alu deletion at locus P01730 were recorded respectively in Dungans ( 0.04 ) , immigrants from China , and Tajiks ( 0.15 ) . The coefficient of gene differentiation in the Central Asian populations for all the genes was 2.8 % , which indicates a relatively low level of population genetic subdivision in this region . The unity of the gene pool of the Central Asian Caucasoids was shown . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy . Concomitant expression of urokinase type plasminogen activator ( uPA ) and its surface receptor ( Q03405 ) has been shown to correlate strongly with a more invasive tumor cell phenotype . A highly malignant human epidermoid carcinoma cell line ( HEp3 ) was transfected with a vector capable of expressing an antisense transcript complementary to 300 bases of the 5' end of Q03405 , including the ATG codon . Six stably transfected antisense ( AS-2 , 3 , 5 , 9 , 10 , 12 ) and eight control clones were characterized . All clones produced high levels of uPA activity . Examination of collagenase production and doubling time showed that all of the clones tested produced similar activities . The antisense clones showed a 20-74 % reduction in the Q03405 sites ; the Q03405 mRNA level was also reduced . A test of the invasive ability of all clones in a modified chorioallantoic membrane ( P62158 ) showed that invasiveness of the antisense-inhibited clones was directly proportional to the density of surface Q03405 . The AS-2 clone , which expressed the lowest number of uPARs showed a significantly reduced level of invasion . The invasiveness of additional AS-inhibited clones was also reduced . Seven control and four AS-inhibited clones were tested for tumorigenicity on CAMs of chick embryos . Inoculation of control cells produced large tumors , while the As clones were non-tumorigenic . AS-2 did not produce tumors even if kept in vivo for up to 10 weeks. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00013 plasminogen activator system gene expression is increased in human breast carcinoma and its bone metastases -- a comparison of normal breast tissue , non-invasive and invasive carcinoma and osseous metastases . The urokinase plasminogen activator ( uPA ) system has been widely associated with the development of breast carcinoma . However , the role of the urokinase pathway in the development of osseous breast cancer metastases has been largely overlooked . We studied the expression of uPA , urokinase plasminogen activator receptor ( Q03405 ) - and plasminogen activator inhibitor type-1 ( P05121 ) in human breast carcinomas and their bone metastases , using in situ hybridisation . Studies were performed using paraffin-embedded tissue from 13 ductal carcinomas , 23 invasive ductal carcinomas , five normal breasts and 25 bone metastases . The majority of the tumours examined expressed low to moderate levels of uPA mRNA and low to high levels of Q03405 and P05121 mRNA , which was predominantly localised to the epithelial tumour cells . There was slight over-expression of uPA and P05121 mRNA and a marked increase in Q03405 mRNA expression in the malignant tumours compared with benign tissue . Overall , Q03405 and P05121 mRNA expression was found to be more variable than uPA mRNA , suggesting a possible role of the receptor and inhibitor in the regulation of uPA activity . Increased alpha1(I) procollagen ( COL ) and osteopontin ( P10451 ) mRNA expression was detected , primarily in the stromal cells , in malignant tumours compared with the benign tissue . The increased expression of the components of the uPA system on the epithelial tumour cells may account for the activation of the proteolytic cascade that occurs during breast cancer metastasis to bone . Furthermore , the over-expression of COL and P10451 suggests a possible interaction between these matrix proteins and the uPA system . DB00877 induces the TGFbeta1/Smad signaling cascade in renal mesangial cells upstream of P42345 . The P42345 kinase inhibitor rapamycin ( sirolimus ) is a drug with potent immunosuppressive and antiproliferative properties . We found that rapamycin induces the TGFbeta/Smad signaling cascade in rat mesangial cells ( MC ) as depicted by the nuclear translocation of phospho-Smads 2 , -3 and Smad-4 , respectively . Concomitantly , rapamycin increases the nuclear DNA binding of receptor ( R ) - and co-Smad proteins to a cognate Smad-binding element ( SBE ) which in turn causes an increase in profibrotic gene expression as exemplified by the connective tissue growth factor ( P29279 ) and plasminogen activator inhibitor 1 ( P05121 ) . Using small interfering (si)RNA we demonstrate that Smad 2/3 activation by rapamycin depends on its endogenous receptor FK binding protein 12 ( P62942 ) . Mechanistically , Smad induction by rapamycin is initiated by an increase in active TGFbeta(1) as shown by ELISA and by the inhibitory effects of a neutralizing TGFbeta antibody . Using an activin receptor-like kinase ( Q9UM73 ) -5 inhibitor and by siRNA against the TGFbeta type II receptor ( TGFbeta-RII ) we furthermore demonstrate a functional involvement of both types of TGFbeta receptors . However , rapamycin did not compete with TGFbeta for TGFbeta-receptor binding as found in radioligand-binding assay . Besides SB203580 , a specific inhibitor of the p38 MAPK , the reactive oxygen species ( ROS ) scavenger N-acetyl-cysteine ( Q9C000 ) and a cell-permeable superoxide dismutase ( SOD ) mimetic strongly abrogated the stimulatory effects of rapamycin on Smad 2 and 3 phosphorylation . Furthermore , the rapid increase in dichlorofluorescein ( DCF ) formation implies that rapamycin mainly acts through ROS . In conclusion , activation of the profibrotic TGFbeta/Smad signaling cascade accompanies the immunosuppressive and antiproliferative actions of rapamycin . Attenuation of plasminogen activator inhibitor type-1 promoter activity in serum-stimulated renal epithelial cells by a distal 5' flanking region . DB00013 plasminogen activator ( u-PA ) and its fast acting type-1 inhibitor ( P05121 ) localize to cellular focal adhesive structures and the adjoining proximal undersurface region , respectively ( Kutz et al. , J. Cell. Physiol. 176:8-18 , 1997 ) . P05121 may function in this locale to modulate pericellular proteolytic activity , cell-to-substrate adhesion , or matrix-dependent motility . While P05121 synthesis is regulated in an immediate-early response manner in growth " activated " renal cells coincident with cytoskeletal restructuring , adhesive influences both repress the amplitude and prolong the time course of serum-induced P05121 transcription ( Ryan et al. , Biochem. J. 314:1041-1046 , 1996 ) . To identify potential adhesion-responsive elements within the P05121 gene that function in this complex mode of expression control , reporter constructs containing defined directionally deleted P05121 5' genomic fragments cloned upstream of a CAT gene were employed in transient transfection assays . A 483-bp distal P05121 flanking segment ( corresponding to nucleotides -2395 to -1912 ) conferred significant adhesion-dependent attenuation on serum-induced P05121 transcription . This 483-bp distal P05121 segment functioned as a repressor of reporter ( CAT ) activity under both adhesive and suspension culture conditions , however , when ligated upstream of either an SV40 promoter/enhancer or a minimal P05121 promoter . These data suggest that repressor elements located between -2395 and -1912 bp interact with more proximal adhesion-dependent regulatory elements to affect P05121 expression attenuation during serum stimulation of adherent renal epithelial cells . DB00013 receptor and resistance to targeted anticancer agents . The urokinase receptor ( Q03405 ) is a P06744 -anchored membrane protein , which regulates protease activity at the cell surface and , in collaboration with a system of co-receptors , triggers cell-signaling and regulates gene expression within the cell . In normal tissues , Q03405 gene expression is limited ; however , in cancer , Q03405 is frequently over-expressed and the gene may be amplified . Hypoxia , which often develops in tumors , further increases Q03405 expression by cancer cells . Q03405 -initiated cell-signaling promotes cancer cell migration , invasion , metastasis , epithelial-mesenchymal transition , stem cell-like properties , survival , and release from states of dormancy . Newly emerging data suggest that the pro-survival cell-signaling activity of Q03405 may allow cancer cells to " escape " from the cytotoxic effects of targeted anticancer drugs . Herein , we review the molecular properties of Q03405 that are responsible for its activity in cancer cells and its ability to counteract the activity of anticancer drugs . Receptor mediated binding of the fibrinolytic components , plasminogen and urokinase , to peripheral blood cells . DB00142 -plasminogen binds to platelets ; the monocytoid line , U937 , and the human fetal fibroblast line , GM1380 bind both plasminogen and its activator , urokinase . This study assesses the interaction of these fibrinolytic proteins with circulating human blood cells . P00747 bound minimally to red cells but bound saturably and reversibly to monocytes , granulocytes and lymphocytes with apparent Kd values of 0.9-1.4 microM . The interactions were of high capacity with 1.6 to 49 X 10(5) sites/cell and involved the lysine binding sites of plasminogen . Both T cells and non-rosetting lymphocytes and two B cell lines saturably bound plasminogen . DB00013 bound saturably to granulocytes , monocytes , non-rosetting lymphocytes and a B cell line , but minimally to T cells , platelets and red cells . Therefore , plasminogen binding sites of high capacity , of similar affinities , and with common recognition specificities are expressed by many peripheral blood cells . DB00013 receptors are also widely distributed , but less so than plasminogen binding sites . The binding of plasminogen and/or urokinase to these cells may lead to generation of cell-associated proteolytic activity which contributes to a variety of cellular functions . DB00013 receptor is necessary for bacterial defense against pneumonia-derived septic melioidosis by facilitating phagocytosis . DB00013 receptor ( urokinase-type plasminogen activator receptor [ Q03405 ] , CD87 ) , a P06744 -anchored protein , is considered to play an important role in inflammation and fibrinolysis . The Gram-negative bacterium Burkholderia pseudomallei is able to survive and replicate within leukocytes and causes melioidosis , an important cause of pneumonia-derived community-acquired sepsis in Southeast Asia . In this study , we investigated the expression and function of Q03405 both in patients with septic melioidosis and in a murine model of experimental melioidosis . Q03405 mRNA and surface expression was increased in patients with septic melioidosis in/on both peripheral blood monocytes and granulocytes as well as in the pulmonary compartment during experimental pneumonia-derived melioidosis in mice . Q03405 -deficient mice intranasally infected with B. pseudomallei showed an enhanced growth and dissemination of B. pseudomallei when compared with wild-type mice , corresponding with increased pulmonary and hepatic inflammation . Q03405 knockout mice demonstrated significantly reduced neutrophil migration toward the pulmonary compartment after inoculation with B. pseudomallei . Further in vitro experiments showed that Q03405 -deficient macrophages and granulocytes display a markedly impaired phagocytosis of B. pseudomallei . Additional studies showed that Q03405 deficiency did not influence hemostatic and fibrinolytic responses during severe melioidosis . These data suggest that Q03405 is crucially involved in the host defense against sepsis caused by B. pseudomallei by facilitating the migration of neutrophils toward the primary site of infection and subsequently facilitating the phagocytosis of B. pseudomallei . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Regulation of the Q03405 /uPA system expressed on monocytes by the deactivating cytokines , P05112 , P22301 and P35225 : consequences on cell adhesion to vitronectin and fibrinogen . DB00013 ( uPA ) and its receptor ( Q03405 ) have been proposed to be involved in monocyte migration by inducing degradation of matrix proteins . In addition , Q03405 is also implicated in cell adhesion to the vascular wall . The adhesive function of Q03405 depends on a direct interaction with vitronectin which is increased by uPA and by modification of cell surface integrin ( such as CD11b- P05107 ) when associated to Q03405 . In this study we analysed the role of three deactivating cytokines , P05112 , P22301 and P35225 , on the surface expression of uPA , Q03405 and CD11b by monocytes and their consequences on monocyte adhesion to immobilized fibrinogen and vitronectin . P22301 induced a decrease in uPA and CD11b after 18 h incubation and a delayed decrease in Q03405 which was only significant after 48 h incubation . These results may explain the decrease in monocyte adhesion , which was observed after an 18 h incubation with P22301 , on immobilized vitronectin and fibrinogen . In contrast , P05112 and P35225 induced a decrease in Q03405 after 18 h and a significant increase in uPA both in the cell lysates and at the cell surface , as well as an increase in cell surface associated CD11b . These cytokines did not modify cell adhesiveness to vitronectin or fibrinogen despite the increase in CD11b- P05107 . This could be due to the decrease in Q03405 because CD11b- P05107 / Q03405 forms a cell adhesion complex . In addition , the increase in uPA induced by P05112 could counterbalance the direct interaction of Q03405 with vitronectin . The increase in uPA suggests that P05112 and P35225 could induce plaque fissuring by monocytes , whereas P22301 may induce protection against matrix protein degradation by decreasing uPA .

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interacts_with DB00674?

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Epigenetic mechanisms in the dopamine D2 receptor-dependent inhibition of the prolactin gene . Transcription of the prolactin gene is dynamically controlled by positive and negative hormone signals that target the regulatory promoter region . Based on the inducibility of prolactin gene expression by inhibitors of histone deacetylases ( HDACs ) , we examined the role of histone acetylation at the genomic prolactin promoter as a late step in transcriptional regulation . Chromatin immunoprecipitation analysis of GH4 cells revealed elevated levels of acetylated histones in the promoter and enhancer regions of the gene , compared with downstream intron sequences . 17beta-Estradiol stimulated histone H4 acetylation in the promoter region by 2- to 3-fold within 30 min . Dopamine inhibited histone H4 acetylation by 2-fold in 30 min , an effect mimicked by the MAPK kinase ( Q02750 ) inhibitor U0126 . In contrast , the synthetic glucocorticoid dexamethasone , which inhibits prolactin transcription , failed to alter histone acetylation over the same time frame . Association of transcription activator Pit-1 with the prolactin promoter was unchanged by hormone treatment . However , in response to dopamine , histone deacetylase Q92769 and corepressor mSin3A were rapidly recruited to the prolactin promoter , and association was sustained above basal levels over a 1-h period . Consistent with this corepressor function , depletion of endogenous mSin3A by small interfering RNA was sufficient to enhance prolactin gene expression by 70 % , comparable to the induction by the HDAC inhibitor , trichostatin A . These studies demonstrate that dopamine D2 receptor activation and inhibition of MAPK ( P27361 /2 ) signaling lead to rapid deacetylation of histones at the genomic prolactin promoter . Recruitment of specific HDAC/ corepressor complexes may be an important mechanism for repression of target gene transcription by Gi/o-coupled receptors . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Different cholinesterase inhibitor effects on P04141 cholinesterases in Alzheimer patients . BACKGROUND : The current study aimed to compare the effects of different cholinesterase inhibitors on acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) activities and protein levels , in the cerebrospinal fluid ( P04141 ) of Alzheimer disease ( AD ) patients . METHODS AND FINDINGS : AD patients aged 50-85 years were randomized to open-label treatment with oral rivastigmine , donepezil or galantamine for 13 weeks . P22303 and BuChE activities were assayed by Ellman 's colorimetric method . Protein levels were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Primary analyses were based on the Completer population ( randomized patients who completed Week 13 assessments ) . 63 patients were randomized to treatment . DB00989 was associated with decreased P22303 activity by 42.6 % and decreased P22303 protein levels by 9.3 % , and decreased BuChE activity by 45.6 % and decreased BuChE protein levels by 21.8 % . DB00674 decreased P22303 activity by 2.1 % and BuChE activity by 0.5 % , but increased P22303 protein levels by 51.2 % and BuChE protein levels by 10.5 % . Donepezil increased P22303 and BuChE activities by 11.8 % and 2.8 % , respectively . Donepezil caused a 215.2 % increase in P22303 and 0.4 % increase in BuChE protein levels . Changes in mean P22303 -Readthrough/Synaptic ratios , which might reflect underlying neurodegenerative processes , were 1.4 , 0.6 , and 0.4 for rivastigmine , donepezil and galantamine , respectively . CONCLUSION : The findings suggest pharmacologically-induced differences between rivastigmine , donepezil and galantamine . DB00989 provides sustained inhibition of P22303 and BuChE , while donepezil and galantamine do not inhibit BuChE and are associated with increases in P04141 P22303 protein levels . The clinical implications require evaluation . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . DB01098 improves endothelial function in db/db mice : role of angiotensin II type 1 receptors and oxidative stress . BACKGROUND AND PURPOSE : P04035 inhibitors , statins , with lipid-reducing properties combat against atherosclerosis and diabetes . The favourable modulation of endothelial function may play a significant role in this effect . The present study aimed to investigate the cellular mechanisms responsible for the therapeutic benefits of rosuvastatin in ameliorating diabetes-associated endothelial dysfunction . EXPERIMENTAL APPROACH : Twelve-week-old db/db diabetic mice were treated with rosuvastatin at 20 mg·kg⁻¹ ·day⁻¹ p.o.for 6 weeks . Isometric force was measured in isolated aortae and renal arteries . Protein expressions including angiotensin II type 1 receptor ( AT₁R ) , Q9NPH5 , O75935 (phox) , p67(phox) , Rac-1 , nitrotyrosine , phospho- P27361 /2 and phospho-p38 were determined by Western blotting , while reactive oxygen species ( ROS ) accumulation in the vascular wall was evaluated by dihydroethidium fluorescence and lucigenin assay . KEY RESULTS : DB01098 treatment of db/db mice reversed the impaired ACh-induced endothelium-dependent dilatations in both renal arteries and aortae and prevented the exaggerated contractions to angiotensin II and phenylephrine in db/db mouse renal arteries and aortae . DB01098 reduced the elevated expressions of AT₁R , O75935 (phox) and p67(phox) , Q9NPH5 , Rac1 , nitrotyrosine and phosphorylation of P27361 /2 and p38 MAPK and inhibited ROS production in aortae from db/db mice . CONCLUSIONS AND IMPLICATIONS : The vasoprotective effects of rosuvastatin are attributed to an increase in NO bioavailability , which is probably achieved by its inhibition of ROS production from the AT₁R-NAD(P)H oxidase cascade . P12821 ( P12821 ) and Q9BYF1 levels in the cerebrospinal fluid of patients with multiple sclerosis . BACKGROUND : We reported a reduction in the levels of angiotensin II in cerebrospinal fluid ( P04141 ) from patients with multiple sclerosis ( MS ) . OBJECTIVE AND METHODS : To clarify the mechanism underlying this reduction , we assayed angiotensin-converting enzyme ( P12821 ) and Q9BYF1 concentrations along with angiotensin II concentrations in P04141 samples from 20 patients with MS and 17 controls with non-neurological diseases . RESULTS : P12821 levels were significantly elevated in patients with MS compared with controls ( 48.42 +/- 4.84 vs 44.71 +/- 3.9 pg/mL ) , whereas Q9BYF1 levels were significantly reduced ( 2.56 +/- 0.26 vs 2.78 +/- 0.24 pg/mL ) , acting toward a normalization of angiotensin II levels . CONCLUSION : These results further indicate an alteration of the intrathecal renin-angiotensin system in patients with MS . Human immunodeficiency virus ( HIV ) -induced neurotoxicity : roles for the DB01221 receptor subtypes . Neuronal damage in human immunodeficiency virus type 1 ( HIV-1 ) infection in the brain is thought to occur at least in part through DB01221 receptor ( NMDAR ) excitation initiated by soluble neurotoxins from HIV-infected brain macrophages . Furthermore , brain regions enriched in NMDAR-2A ( Q12879 ) and NMDAR-2B ( Q13224 ) subunits , such as the hippocampus , are particularly vulnerable . Using cultured rat hippocampal cells and HIV-1-infected human monocyte-derived macrophages ( HIV/MDM ) , we examined the role of Q12879 and Q13224 in HIV/MDM-induced hippocampal neuronal death . We used the primary HIV-1 strain Jago derived from the P04141 of an individual with HIV-associated dementia and that robustly replicates in MDM . We found the following : ( 1 ) hippocampal neuronal susceptibility to HIV/MDM excitotoxins varies according to the developmental expression patterns of Q12879 and Q13224 ; ( 2 ) NMDAR activation by HIV/MDM results in neuronal calpain activation , which results in neuronal death ; and ( 3 ) selective antagonists of homomeric Q13224 / Q13224 - and heteromeric Q12879 / Q13224 -containing NMDARs , as well as an inhibitor of calpain activity , afford neuroprotection against HIV/MDM . These studies establish a clear link between macrophage HIV infection , neuronal Q12879 and Q13224 activation , and calpain-mediated hippocampal neuronal death . They further suggest a dominant role for Q12879 and Q13224 in determining neuronal susceptibility in HIV-infected brain . Antagonists of Q12879 and Q13224 subunits as well as inhibitors of calpain activation offer attractive neuroprotective approaches against HIV in both developing and mature brain . Mobilization of Ph chromosome-negative peripheral blood stem cells in chronic myeloid leukaemia patients with imatinib mesylate-induced complete cytogenetic remission . Imatinib mesylate ( IM , DB00619 , Glivec ) can induce a high rate of complete cytogenetic response ( CCR ) in chronic myeloid leukaemia ( CML ) patients , although to date the majority of patients continue to have detectable disease by sensitive reverse transcription polymerase chain reaction ( RT-PCR ) . It is therefore possible that these patients may ultimately relapse and require treatment such as autologous peripheral blood stem cell transplant ( APBSCT ) . We attempted mobilization of haemopoietic progenitor cells from 58 patients in CCR using recombinant human granulocyte colony-stimulating factor [ rHu- DB00099 ; 10 micro g/kg/d subcutaneously ( s.c. ) for at least 4 d ] alone , while continuing IM treatment . The median d 5 ( peak ) P28906 + count was 11.5/ microl ( range 0-108/ microl ) , and 43/58 ( 74 % ) patients underwent a median of two ( range 1-3 ) apheresis procedures . A median dose of 2.1 x 10(6)/kg P28906 + cells ( range 0.1-6.5 x 10(6)/kg ) was collected . Some 84 % of 31 collections analysed were negative for the Philadelphia ( Ph ) chromosome or breakpoint cluster region and Abelson murine leukaemia viral oncogene homologue ( P11274 - P00519 ) translocation by cytogenetics or fluorescent in situ hybridization respectively . No toxicity was reported with the regimen . Overall , the target P28906 + dose ( 2 x 10(6)/kg P28906 + ) was attained in 23/58 ( 40 % ) patients who entered the study . In summary , we have demonstrated that successful mobilization of Ph- P28906 + cells from IM-treated patients in CCR is possible using rHu-G- P04141 alone . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Effect of endothelin receptor antagonists on non-muscle matrix compaction in a cell culture vasospasm model . P05305 ( ET-1 ) , a potent vascular smooth muscle constrictor , is one of the possible spasmogens in cerebral vasospasm . However , the role of ET-1 in non-muscle compaction ( another aspect of the pathogenesis of cerebral vasospasm ) has not been reported . This study was undertaken to demonstrate the effect of ET-1 , as well as erythrocyte lysate and bloody cerebrospinal fluid ( P04141 ) , on fibroblast populated collagen lattice ( FPCL ) compaction . Human dermal fibroblasts were used to form FPCL . The concentration-dependent effect of ET-1 was examined in the absence and presence of an P25101 receptor antagonist ( BQ-485 ) , or an ETB receptor antagonist ( BQ-788 ) , or both . FPCL compaction was determined by measuring reduction of areas over five days following treatment . To compare the effect of ET-1 on lattice compaction , erythrocyte lysate and bloody P04141 obtained from a cerebral vasospasm patient were also tested . We found that ET-1 increased FPCL compaction in a concentration-dependent ( but not time-dependent ) manner . Erythrocyte lysate produced the strongest compaction , however , without time-dependence . Bloody P04141 promoted FPCL compaction in a time-dependent fashion . Compaction induced by ET-1 was inhibited by BQ-485 but not by BQ-788 . We concluded that ET-1 promotes FPCL compaction by activation of P25101 receptors . Other components in bloody P04141 or erythrocytes may also contribute to FPCL compaction . Altered endothelin receptor expression and affinity in spontaneously hypertensive rat cerebral and coronary arteries . BACKGROUND : Hypertension is associated with arterial hyperreactivity , and endothelin ( ET ) receptors are involved in vascular pathogenesis . The present study was performed to examine the hypothesis that ET receptors were altered in cerebral and coronary arteries of spontaneously hypertensive rats ( SHR ) . METHODOLOGY/PRINCIPAL FINDINGS : Cerebral and coronary arteries were removed from SHR . Vascular contraction was recorded using a sensitive myograph system . Real-time PCR and Western blotting were used to quantify mRNA and protein expression of receptors and essential MAPK pathway molecules . The results demonstrated that both P25101 and ETB receptor-mediated contractile responses in SHR cerebral arteries were shifted to the left in a nonparallel manner with increased maximum contraction compared with Wistar-Kyoto ( WKY ) rats . In SHR coronary arteries , the P25101 receptor-mediated contraction curve was shifted to the left in parallel with an increased pEC50 compared with the arteries in WKY rats . There was no significant increase in ETB receptor-mediated contraction in SHR coronary arteries . P25101 receptor mRNA and protein expression was increased in SHR cerebral arteries compared with the arteries in WKY rats . However , P25101 receptor mRNA and protein levels in coronary arteries and ETB receptor protein levels in cerebral and coronary arteries remained unchanged in SHR compared with WKY rats . Meanwhile , phosphorylated P27361 /2 protein was significantly increased in SHR brain and heart vessels . CONCLUSIONS/SIGNIFICANCE : In SHR cerebral arteries , P25101 receptor expression was upregulated . P25101 receptor affinity was increased in coronary arteries , and ETB receptor affinity was increased in cerebral arteries . The P27361 /2 activation may be involved in the receptor alterations . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . DB00674 ameliorates the impairment of recognition memory in mice repeatedly treated with methamphetamine : involvement of allosteric potentiation of nicotinic acetylcholine receptors and dopaminergic- P27361 /2 systems . DB00674 , a drug used to treat Alzheimer 's disease , inhibits acetylcholinesterase ( P22303 ) and allosterically modulates nicotinic acetylcholine receptors ( nAChRs ) resulting in stimulation of catecholamine neurotransmission . In this study , we investigated whether galantamine exerts cognitive-improving effects through the allosteric modulation of nAChRs in an animal model of methamphetamine ( Meth ) psychosis . The mice treated with Meth ( 1 mg/kg.d ) for 7 d showed memory impairment in a novel object recognition test . DB00674 ( 3 mg/kg ) ameliorated the memory impairment , and it increased the extracellular dopamine release in the prefrontal cortex ( P27918 ) of Meth-treated mice . Donepezil , an P22303 inhibitor ( 1 mg/kg ) increased the extracellular ACh release in the P27918 , whereas it had no effect on the memory impairment in Meth-treated mice . The nAChR antagonist , mecamylamine , and dopamine D1 receptor antagonist , P35240 23390 , blocked the ameliorating effect of galantamine on Meth-induced memory impairment , whereas the muscarinic AChR antagonist , scopolamine , had no effect . The effects of galantamine on extracellular dopamine release were also antagonized by mecamylamine . DB00674 attenuated the defect of the novelty-induced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . The ameliorating effect of galantamine on recognition memory in Meth-treated mice was negated by microinjection of an P29323 inhibitor , PD98059 , into the P27918 . These results suggest that the ameliorating effect of galantamine on Meth-induced memory impairment is associated with indirect activation of dopamine D1 receptor- P27361 /2 following augmentation with dopaminergic neurotransmission in the P27918 through the allosteric activation of nAChRs . DB00674 could be a useful therapeutic agent for treating cognitive deficits in schizophrenia/Meth psychosis , as well as Alzheimer 's disease . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . Transforming growth factor alpha-induced expression of type 1 plasminogen activator inhibitor in astrocytes rescues neurons from excitotoxicity . Although transforming growth factor ( TGF ) -alpha , a member of the epidermal growth factor ( P01133 ) family , has been shown to protect neurons against excitotoxic and ischemic brain injuries , its mechanism of action remains unknown . In the present study , we used in vitro models of apoptotic or necrotic paradigms demonstrating that TGF-alpha rescues neurons from N-methyl-D-aspartate ( DB01221 ) -induced excitotoxic death , with the obligatory presence of astrocytes . Because neuronal tissue-type plasminogen activator ( t-PA ) release was shown to potentiate DB01221 -induced excitotoxicity , we observed that TGF-alpha treatment reduced DB01221 -induced increase of t-PA activity in mixed cultures of neurons and astrocytes . In addition , we showed that although TGF-alpha induces activation of the extracellular signal-regulated kinases ( ERKs ) in astrocytes , it failed to activate Q8NFH3 / Q8TCB0 in neurons . Finally , we showed that TGF-alpha , by an P29323 -dependent mechanism , stimulates the astrocytic expression of P05121 , a t-PA inhibitor , which mediates the neuroprotective activity of TGF-alpha against DB01221 -mediated excitotoxic neuronal death . Taken together , we indicate that TGF-alpha rescues neurons from DB01221 -induced excitotoxicity in mixed cultures through inhibition of t-PA activity , involving P05121 overexpression by an P29323 -dependent pathway in astrocytes . Phosphodiesterase 4B mediates extracellular signal-regulated kinase-dependent up-regulation of mucin P98088 protein by Streptococcus pneumoniae by inhibiting DB02527 -protein kinase A-dependent P28562 phosphatase pathway . Otitis media ( OM ) is the most common childhood bacterial infection and the major cause of conductive hearing loss in children . Mucus overproduction is a hallmark of OM . Streptococcus pneumoniae is the most common gram-positive bacterial pathogen causing OM . Among many mucin genes , P98088 has been found to be greatly up-regulated in the middle ear mucosa of human patients with OM . We previously reported that S. pneumoniae up-regulates P98088 expression in a MAPK P29323 -dependent manner . We also found that MAPK phosphatase-1 ( P28562 ) negatively regulates S. pneumoniae-induced P29323 -dependent P98088 up-regulation . Therapeutic strategies for up-regulating the expression of negative regulators such as P28562 may have significant therapeutic potential for treating mucus overproduction in OM . However , the underlying molecular mechanism by which P28562 expression is negatively regulated during S. pneumoniae infection is unknown . In this study we show that phosphodiesterase 4B ( Q07343 ) mediates S. pneumoniae-induced P98088 up-regulation by inhibiting the expression of a negative regulator P28562 , which in turn leads to enhanced MAPK P29323 activation and subsequent up-regulation of P98088 . Q07343 inhibits P28562 expression in a DB02527 -PKA-dependent manner . DB05876 -specific inhibitor rolipram inhibits S. pneumoniae-induced P98088 up-regulation both in vitro and in vivo . Moreover , we show that Q07343 plays a critical role in P98088 induction . Finally , topical and post-infection administration of rolipram into the middle ear potently inhibited S. pneumoniae-induced P98088 up-regulation . Collectively , these data demonstrate that Q07343 mediates P29323 -dependent up-regulation of mucin P98088 by S. pneumoniae by inhibiting DB02527 -PKA-dependent P28562 pathway . This study may lead to novel therapeutic strategy for inhibiting mucus overproduction . P04150 recruitment of histone deacetylase 2 inhibits interleukin-1beta-induced histone H4 acetylation on lysines 8 and 12 . We have investigated the ability of dexamethasone to regulate interleukin-1beta ( IL-1beta ) -induced gene expression , histone acetyltransferase ( O60235 ) and histone deacetylase ( HDAC ) activity . Low concentrations of dexamethasone ( 10(-10) M ) repress IL-1beta-stimulated granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) expression and fail to stimulate secretory leukocyte proteinase inhibitor expression . Dexamethasone ( 10(-7) M ) and IL-1beta ( 1 ng/ml ) both stimulated O60235 activity but showed a different pattern of histone H4 acetylation . Dexamethasone targeted lysines P13647 and P08779 , whereas IL-1beta targeted K8 and K12 . Low concentrations of dexamethasone ( 10(-10) M ) , which do not transactivate , repressed IL-1beta-stimulated K8 and K12 acetylation . Using chromatin immunoprecipitation assays , we show that dexamethasone inhibits IL-1beta-enhanced acetylated K8-associated GM- P04141 promoter enrichment in a concentration-dependent manner . Neither IL-1beta nor dexamethasone elicited any GM- P04141 promoter association at acetylated P13647 residues . Furthermore , we show that GR acts both as a direct inhibitor of CREB binding protein ( CBP ) -associated O60235 activity and also by recruiting Q92769 to the p65-CBP O60235 complex . This action does not involve de novo synthesis of HDAC protein or altered expression of CBP or p300/CBP-associated factor . This mechanism for glucocorticoid repression is novel and establishes that inhibition of histone acetylation is an additional level of control of inflammatory gene expression . This further suggests that pharmacological manipulation of of specific histone acetylation status is a potentially useful approach for the treatment of inflammatory diseases . A Type II Arabinogalactan from Anoectochilus formosanus for G- P04141 Production in Macrophages and Leukopenia Improvement in Q86TM3 -Bearing Mice Treated with DB00544 . Anoectochilus formosanus is an herb well known in Asian countries . The polysaccharide isolated from A. formosanus consists of type II arabinogalactan ( AGAF ) , with branched 3,6-Gal as the major moiety . In this study , AGAF was examined for the granulocyte colony-stimulating factor ( DB00099 ) production and related protein expression in RAW 264.7 murine macrophages . The signaling pathway of G- P04141 production involves AGAF and mitogen-activated protein kinases ( MAPKs ) inhibitors and pattern-recognition receptor antibodies . AGAF was evaluated to ease the leukopenia in Q86TM3 -colon-cancer-bearing mice treated with 5-fluorouracil ( DB00544 ) . The results of this study showed that AGAF was a stimulant for O60603 and Q9BXN2 and that it induced G- P04141 production , through p38 and P29323 MAPK , and NF- κ B pathways . In vivo examination showed that the oral administration of AGAF mitigated the side effects of leukopenia caused by DB00544 in colon-cancer-bearing mice . In conclusion , the botanic type II AGAF in this study was a potent G- P04141 inducer in vivo and in vitro . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . P28482 , but not P27361 , mediates acquired and " de novo " resistance to imatinib mesylate : implication for CML therapy . Resistance to Imatinib Mesylate ( IM ) is a major problem in Chronic Myelogenous Leukaemia management . Most of the studies about resistance have focused on point mutations on P11274 / P00519 . However , other types of resistance that do not imply mutations in P11274 / P00519 have been also described . In the present report we aim to study the role of several MAPK in IM resistance not associate to P11274 / P00519 mutations . Therefore we used an experimental system of resistant cell lines generated by co-culturing with IM ( K562 , Lama 84 ) as well as primary material from resistant and responder patient without P11274 / P00519 mutations . Here we demonstrate that Erk5 and p38MAPK signaling pathways are not implicated in the acquired resistance phenotype . However , Erk2 , but not Erk1 , is critical for the acquired resistance to IM . In fact , Bcr/Abl activates preferentially Erk2 in transient transfection in a dose dependent fashion through the c-Abl part of the chimeric protein . Finally , we present evidences demonstrating how constitutive activation of Erk2 is a de novo mechanism of resistance to IM . In summary our data support the use of therapeutic approaches based on Erk2 inhibition , which could be added to the therapeutic armamentarium to fight CML , especially when IM resistance develops secondary to Erk2 activation . G- P04141 increases secretion of urokinase-type plasminogen activator by human lung cancer cells . We reported previously that granulocyte colony-stimulating factor ( DB00099 ) can promote the invasion of human lung cancer cell lines in vitro . However , the exact mechanism of its stimulatory effect on invasion remains to be elucidated . In the present study we mainly focused our attention on the components of the plasminogen activation system in human lung cancer cell lines , because of the central role that plasminogen activators play in regulating extracellular proteolysis . We showed that G- P04141 induced a dose-dependent increase in the urokinase-type plasminogen activator ( uPA ) activity in the conditioned medium of a PC-9 lung cancer cell line . When the amounts of uPA activity were quantitated by densitometry , we found that even at a concentration of 0.01 microg/ml , G- P04141 had a stimulatory effect on the uPA release , while high concentrations caused a 3.6-fold increase at a maximum concentration of 1 microg/ml . A Western blot analysis of the conditioned medium confirmed the findings observed in a zymographic analysis . The observed increase in uPA protein was paralleled by a significant increase in the uPA mRNA levels after treatment with DB00099 . However , our experiments failed to identify any alteration in the plasminogen activator inhibitor ( P05121 ) secretion caused by DB00099 . In addition , we also found the expression of Q99062 by PC-9 cells , suggesting the possible pathway activated by DB00099 . Increased miR-21 expression during human monocyte differentiation into DCs . Differentiation of monocytes into dendritic cells ( DCs ) is characterised by marked changes in gene expression . The role of microRNAs ( miRNAs ) , a new class of small endogenous non-coding regulatory RNAs , in this process is still unclear . We identified miR-223 , miR-16 , miR-191 , miR-24 , let-7b , and miR-21 as differentially expressed between monocytes and monocyte derived DCs . We evaluated the expression levels of computationally predicted target genes of miR-21 in human monocytes following stimulation with GM- P04141 and P05112 . Moreover , transfection of monocytes with synthetic miR-21 inhibited the expression of a set of genes that were also repressed during monocyte differentiation to DCs in response to GM- P04141 and P05112 . Among these , we identified genes that are involved in cell cycle , apoptosis and differentiation such as Q07343 , Q53EL6 , P04083 , Q9NXR8 , Q8N3U4 , Q15582 , P80511 , LAT2 and P48552 . Collectively , the present study highlights the involvement of miRNAs , particularly miR-21 in monocyte differentiation to DCs and identifies potential miR-21 target genes . Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 , P05231 , and P13500 by Cbl-b(-/-) mast cells compared with levels produced by c-Cbl(-/-) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 .

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Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . P05231 sensitizes prostate cancer to the antiproliferative effect of IFNα2 through Q00978 . Development and progression of prostate cancer ( PCa ) are associated with chronic inflammation . The cytokine interleukin 6 ( P05231 ) can influence progression , differentiation , survival , and angiogenesis of PCa . To identify novel pathways that are triggered by P05231 , we performed a gene expression profiling of two PCa cell lines , LNCaP and MDA PCa 2b , treated with 5 ng/ml P05231 . Interferon ( IFN ) regulatory factor 9 ( Q00978 ) was identified as one of the most prevalent P05231 -regulated genes in both cell lines . Q00978 is a mediator of type I IFN signaling and acts together with P42224 and 2 to activate transcription of IFN-responsive genes . The P05231 regulation of Q00978 was confirmed at mRNA and protein levels by quantitative real-time PCR and western blot respectively in both cell lines and could be blocked by the anti- P05231 antibody DB09036 . Three PCa cell lines , PC3 , Du-145 , and LNCaP- P05231 + , with an autocrine P05231 loop displayed high expression of Q00978 . A tissue microarray with 36 PCa tissues showed that Q00978 protein expression is moderately elevated in malignant areas and positively correlates with the tissue expression of P05231 . Downregulation and overexpression of Q00978 provided evidence for an IFN-independent role of Q00978 in cellular proliferation of different PCa cell lines . Furthermore , expression of Q00978 was essential to mediate the antiproliferative effects of IFNα2 . We concluded that P05231 is an inducer of Q00978 expression in PCa and a sensitizer for the antiproliferative effects of IFNα2 . Q13261 drives antagonistic mechanisms of cancer development and immune control in lymphocyte-enriched triple-negative breast cancers . Despite its aggressive nature , triple-negative breast cancer ( TNBC ) often exhibits leucocyte infiltrations that correlate with favorable prognosis . In this study , we offer an explanation for this apparent conundrum by defining TNBC cell subsets that overexpress the P40933 immune receptor Q13261 . This receptor usually forms a heterotrimer with the P60568 receptors P14784 and P31785 , which regulates the proliferation and differentiation of cytotoxic T cells and NK cells . However , unlike Q13261 , the P14784 and P31785 receptors are not upregulated in basal-like TNBC breast cancer cells that express Q13261 . Mechanistic investigations indicated that Q13261 signaling activated P23458 , P42224 , P52630 , AKT , Q96B36 , and P27361 /2 in the absence of P14784 and P31785 , whereas neither P42229 nor O60674 were activated . RNAi-mediated attenuation of Q13261 established its role in cell growth , apoptosis , and migration , whereas expression of the P40933 cytokine in Q13261 -expressing cells stimulated an autocrine signaling cascade that promoted cell proliferation and migration and blocked apoptosis . Notably , coexpression of Q13261 and P40933 was also sufficient to activate peripheral blood mononuclear cells upon coculture in a paracrine signaling manner . Overall , our findings offer a mechanistic explanation for the paradoxical association of some high-grade breast tumors with better survival outcomes , due to engagement of the immune stroma . Topical glucocorticoids downregulate P23219 positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 ) are key enzymes in the synthesis of both pro- ( P23219 , P35354 ) and anti-inflammatory prostanoids ( P35354 ) , we investigated the role of topical GC on P23219 , P35354 and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 , CD117 , MBP , elastase , IgE , BB-1 , P05112 , P05113 and P05231 ) , P23219 and P35354 was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 + cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 + NP cells ( P < 0.05 ) , but have no significant effect on P35354 + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 + cells is however increased significantly upon GC treatment ( P < 0.05 ) . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified . Targeting interleukin-6 in inflammatory autoimmune diseases and cancers . P05231 ( P05231 ) is a pleiotropic cytokine with significant functions in the regulation of the immune system . As a potent pro-inflammatory cytokine , P05231 plays a pivotal role in host defense against pathogens and acute stress . However , increased or deregulated expression of P05231 significantly contributes to the pathogenesis of various human diseases . Numerous preclinical and clinical studies have revealed the pathological roles of the P05231 pathway in inflammation , autoimmunity , and cancer . Based on the rich body of studies on biological activities of P05231 and its pathological roles , therapeutic strategies targeting the P05231 pathway are in development for cancers , inflammatory and autoimmune diseases . Several anti- P05231 / P05231 receptor monoclonal antibodies developed for targeted therapy have demonstrated promising results in both preclinical studies and clinical trials . DB06273 , an anti- P05231 receptor antibody , is effective in the treatment of various autoimmune and inflammatory conditions notably rheumatoid arthritis . It is the only P05231 pathway targeting agent approved by the regulatory agencies for clinical use . DB09036 , an anti- P05231 antibody , has been shown to have potential benefits treating various human cancers either as a single agent or in combination with other chemotherapy drugs . Several other anti- P05231 -based therapies are also under clinical development for various diseases . P05231 antagonism has been shown to be a potential therapy for these disorders refractory to conventional drugs . New strategies , such as combination of P05231 blockade with inhibition of other signaling pathways , may further improve P05231 -targeted immunotherapy of human diseases . P23458 activates P40763 activity in non-small-cell lung cancer cells and P05231 neutralizing antibodies can suppress P23458 - P40763 signaling . Members of the signal transducer and activator of transcription ( P35610 ) family of transcription factors are potential targets for the treatment and prevention of cancers including non-small-cell lung cancer . P35610 proteins can be phosphorylated and activated by diverse upstream kinases including cytokine receptors and tyrosine kinases . We examined P35610 protein activation in lung cancer cell lines including those with activating mutations in the P00533 and examined upstream kinases responsible for P40763 phosphorylation and activation using small molecules , antibodies , and RNA interference . We found more pronounced P40763 activation in cells with activating P00533 mutations , yet inhibition of P00533 activity had no effect on P40763 activation . Inhibition of P23458 with small molecules or RNA interference resulted in loss of P40763 tyrosine phosphorylation and inhibition of cell growth . An interleukin-6 neutralizing antibody , siltuximab ( CNTO 328 ) could inhibit P40763 tyrosine phosphorylation in a cell-dependent manner . DB09036 could completely inhibit P40763 tyrosine phosphorylation in H1650 cells , and this resulted in inhibition of lung cancer cell growth in vivo . Combined P00533 inhibition with erlotinib and siltuximab resulted in dual inhibition of both tyrosine and serine P40763 phosphorylation , more pronounced inhibition of P40763 transcriptional activity , and translated into combined effects on lung cancer growth in a mouse model . Our results suggest that P23458 is responsible for P40763 activation in lung cancer cells and that indirect attacks on P23458 - P40763 using an P05231 neutralizing antibody with or without P00533 inhibition can inhibit lung cancer growth in lung cancer subsets . Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma . Signalling through the interleukin ( IL ) -6 pathway induces proliferation and drug resistance of multiple myeloma cells . We therefore sought to determine whether the P05231 -neutralizing monoclonal antibody siltuximab , formerly CNTO 328 , could enhance the activity of melphalan , and to examine some of the mechanisms underlying this interaction . DB09036 increased the cytotoxicity of melphalan in KAS-6/1 , Q16352 -6 , ANBL-6 , and RPMI 8226 human myeloma cell lines ( HMCLs ) in an additive-to-synergistic manner , and sensitized resistant RPMI 8226.LR5 cells to melphalan . These anti-proliferative effects were accompanied by enhanced activation of drug-specific apoptosis in HMCLs grown in suspension , and in HMCLs co-cultured with a human-derived stromal cell line . DB09036 with melphalan enhanced activation of caspase-8 , caspase-9 , and the downstream effector caspase-3 compared with either of the single agents . This increased induction of cell death occurred in association with enhanced Bak activation . Neutralization of P05231 also suppressed signalling through the phosphoinositide 3-kinase/Akt pathway , as evidenced by decreased phosphorylation of Akt , P08133 S6 kinase and Q13541 . Importantly , the siltuximab/melphalan regimen demonstrated enhanced anti-proliferative effects against primary plasma cells derived from patients with myeloma , monoclonal gammopathy of undetermined significance , and amyloidosis . These studies provide a rationale for translation of siltuximab into the clinic in combination with melphalan-based therapies . Dose selection of siltuximab for multicentric Castleman 's disease . PURPOSE : DB09036 is a monoclonal antibody that binds to interleukin ( IL ) -6 with high affinity and specificity ; P02741 ( CRP ) is an acute-phase protein induced by P05231 . CRP suppression is an indirect measurement of P05231 activity . Here , modeling and simulation of the pharmacokinetic ( PK ) /pharmacodynamic ( PD ) relationship between siltuximab and CRP were used to support dose selection for multicentric Castleman 's disease ( CD ) . METHODS : PK/PD modeling was applied to explore the relationship between siltuximab PK and CRP suppression following intravenous siltuximab infusion in 47 patients with B cell non-Hodgkin 's lymphoma ( n = 17 ) , multiple myeloma ( n = 13 ) , or CD ( n = 17 ) . DB09036 was administered as 2.8 , 5.5 , or 11 mg/kg q2wks , 11 mg/kg q3wks , or 5.5 mg/kg weekly . Simulations of studied or hypothetical siltuximab dosage regimens ( 15 mg/kg q4wks ) were also performed to evaluate maintenance of CRP suppression below the cutoff value of 1 mg/L . RESULTS : A two-compartment PK model and an inhibitory indirect response PD model adequately described the serum siltuximab and CRP concentration-time profiles simultaneously . PD parameter estimates were physiologically plausible . For all disease types , simulations showed that 11 mg/kg q3wks or 15 mg/kg q4wks would reduce serum CRP to below 1 mg/L after the second dose and throughout the treatment period . CONCLUSIONS : PK/PD modeling was used to select doses for further development of siltuximab in multicentric CD . The dosing recommendation was also supported by the observed efficacy dose-response relationship . CRP suppression in the subsequent randomized multicentric CD study was in agreement with the modeling predictions . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . Effects of siltuximab on the P05231 -induced signaling pathway in ovarian cancer . PURPOSE : To explore potential therapeutic strategies for interrupting the interleukin-6 ( P05231 ) signaling pathway , we measured P05231 expression in ovarian cancer tissues , and evaluated the effects of a monoclonal anti- P05231 antibody ; siltuximab ( CNTO 328 ) , on levels of P05231 -induced Stat3 phosphorylation , Stat3 nuclear translocation , and Stat3 downstream antiapoptotic genes . We then looked for enhancing paclitaxel sensitivity in multidrug-resistant ovarian cancer cell lines . EXPERIMENTAL DESIGN : Expressions of P05231 in ovarian cancer patient specimens were assessed by immunohistochemistry . Effects of siltuximab on P05231 -induced activation of Stat3 in an ovarian cancer cell line were determined by Western blot and real-time analysis of Stat3 nucleocytoplasmic translocation . Influence of combination of siltuximab and paclitaxel on tumor growth was evaluated in a xenograft mouse mode in vivo . RESULTS : Metastatic and drug-resistant recurrent tumors have significantly higher P05231 expression when compared with the matched primary tumors . DB09036 specifically suppressed P05231 -induced Stat3 phosphorylation and Stat3 nuclear translocation . Treatment with siltuximab significantly decreased the levels of Stat3 downstream proteins such as Q8WXI8 -1 , Bcl-X(L) , and survivin . Treatment with siltuximab reduced expression of multiple P05231 -induced genes in these cell lines . Furthermore , siltuximab increased the cytotoxic effects of paclitaxel in a paclitaxel resistant ovarian cancer cell line in vitro , but combination therapy with siltuximab did not have a significant effect on paclitaxel resistant tumor growth in vivo . CONCLUSIONS : These results show that siltuximab effectively block the P05231 signaling pathways and P05231 -induced gene expression . Blockage of P05231 signaling may provide benefits for the treatment of ovarian cancer . Activation of serotonin receptor-2B rescues small-for-size liver graft failure in mice . The implantation of grafts below 30 % of the normal liver volume is associated with a high risk of failure known as small-for-size ( SFS ) syndrome . Strategies to rescue small grafts may have a dramatic impact on organ shortage . Serotonin is a potent growth factor for the liver . The goal of this study was to determine whether enhanced serotonin signaling could prevent the deleterious effects of SFS syndrome . We performed 30 % normal liver volume transplantations in wild-type C57/BL6 and interleukin-6 ( P05231 ) (-/-) mice . Some animals received α-methyl-5-HT ( DOI ) , an agonist of serotonin receptor-2 ( P41595 ) . Endpoints included long-term survival , serum and hepatic markers of liver injury and regeneration , assessment of hepatic microcirculation by intravital fluorescence microscopy and scanning electron microscopy , and transcript levels of a variety of serotonin receptors , tumor necrosis factor α , and P05231 . All recipients of small grafts ( controls ) died within 2-4 days of transplantation , whereas half of those receiving DOI survived permanently . Control animals disclosed major liver injury , including diffuse microvesicular steatosis in hepatocytes , impairment of microcirculation , and a failure of regeneration , whereas these parameters were dramatically improved in animals subjected to DOI . Blockage of P41595 blunted the protective effects of DOI . Whereas P05231 levels were higher in DOI-treated animals , P05231 (-/-) mice were still protected by DOI , suggesting a protective pathway independent of P05231 . CONCLUSION : Serotonin through its action on receptor-2B protects SFS liver grafts from injury and prevents microcirculation and regeneration . The mechanism of hepato-protection is independent of P05231 . A phase 2 , randomized , double-blind , placebo-controlled study of siltuximab ( anti- P05231 mAb ) and bortezomib versus bortezomib alone in patients with relapsed or refractory multiple myeloma . We compared the safety and efficacy of siltuximab ( S ) , an anti-interleukin-6 chimeric monoclonal antibody , plus bortezomib ( B ) with placebo ( plc ) + B in patients with relapsed/refractory multiple myeloma in a randomized phase 2 study . DB09036 was given by 6 mg/kg IV every 2 weeks . On progression , B was discontinued and high-dose dexamethasone could be added to S/plc . Response and progression-free survival ( PFS ) were analyzed pre-dexamethasone by European Group for Blood and Marrow Transplantation ( EBMT ) criteria . For the 281 randomized patients , median PFS for S + B and plc + B was 8.0 and 7.6 months ( HR 0.869 , P = 0.345 ) , overall response rate was 55 versus 47 % ( P = 0.213 ) , complete response rate was 11 versus 7 % , and median overall survival ( OS ) was 30.8 versus 36.8 months ( HR 1.353 , P = 0.103 ) . Sustained suppression of P02741 , a marker reflective of inhibition of interleukin-6 activity , was seen with S + B . DB09036 did not affect B pharmacokinetics . DB09036 /placebo discontinuation ( 75 versus 66 % ) , grade ≥3 neutropenia ( 49 versus 29 % ) , thrombocytopenia ( 48 versus 34 % ) , and all-grade infections ( 62 versus 49 % ) occurred more frequently with S + B . The addition of siltuximab to bortezomib did not appear to improve PFS or OS despite a numerical increase in response rate in patients with relapsed or refractory multiple myeloma . Glioma cell activation by Alzheimer 's peptide Abeta1-42 , alpha1-antichymotrypsin , and their mixture . We compared the effects ofAlzheimer 's peptide ( Abeta1-42 ) , a,-antichymotrypsin ( ACT ) and an ACT/Abeta1-42 mixture on human glioma DK-MG cells . The solution of Abeta ( 5 microM ) formed by 2-h incubation at room temperature induced tumour necrosis factor-alpha ( P01375 ) and interleukin ( IL ) -6 levels by 55 and 45 % , respectively , and increased gelatinase B activity by 67 % , while exposure of cells to the ACT/Abeta1-42 mixture ( 1:10 molar ratio ACT : Abeta1-42 ) under the same experimental conditions showed no effect on P05231 levels or gelatinase B activity , but strongly induced P01375 ( by 190 % ) , compared to the controls . Stimulation of the cells with Abeta1-42 alone , but not with ACT , increased by about 20 % low-density lipoprotein ( LDL ) uptake and mRNA levels for P01130 and P04035 , while the ACT/Abeta1-42 mixture significantly increased LDL uptake ( by 50 % ) , up-regulated mRNA levels for P01130 and P04035 by 48 and 63 % , respectively , and increased lipid accumulation by about 20-fold . These data suggest a possible new role for Abeta in Alzheimer 's disease through its interaction with the inflammatory reactant , ACT . P25116 -mediated synovial proliferation in patients with rheumatoid arthritis . Synovial cell proliferation is one of the pathological bases of rheumatoid arthritis ( RA ) . Several cytokines including IL-1 and P05231 and growth factors have been shown to be involved in the synovial cell proliferation in RA . Thrombin is a multifunctional protease and acts as a mitogen for several cell types through its specific receptor . To assess whether thrombin is involved in overproliferation of rheumatoid synovial cells , we measured the concentration of thrombin-anti-thrombin III ( P01008 ) complex ( TAT ) in synovial fluid obtained from patients with RA or osteoarthritis ( OA ) . We also examined the effect of thrombin or thrombin receptor agonist peptide ( TRAP ) on cell growth of synovial cell clones ( SCCs ) established from an RA patient . The concentrations of TAT in the synovial fluid from patients with RA were significantly higher than in those with OA . Moreover , both thrombin and TRAP enhanced proliferation of synovial cells in vitro . We also characterized the expression of thrombin receptor mRNA by reverse transcription-PCR . The expression of mRNA for thrombin receptor was up-regulated by thrombin or TRAP stimulation . P25116 antigen was also detected on both SCCs and synovial tissue from RA patients by immunostaining using a monoclonal antibody against thrombin receptor . These findings indicate that thrombin may act as a mitogen for synovial cells through thrombin receptor and may play some role in synovial overproliferation and remodeling in RA . Neisseria meningitidis capsular polysaccharides induce inflammatory responses via O60603 and O00206 -MD-2 . CPS are major virulence factors in infections caused by Neisseria meningitidis and form the basis for meningococcal serogroup designation and protective meningococcal vaccines . CPS polymers are anchored in the meningococcal outer membrane through a 1,2-diacylglycerol moiety , but the innate immunostimulatory activity of CPS is largely unexplored . Well-established human and murine macrophage cell lines and P29320 /TLR stably transfected cells were stimulated with CPS , purified from an endotoxin-deficient meningococcal serogroup B NMB-lpxA mutant . CPS induced inflammatory responses via O60603 - and O00206 -MD-2 . Meningococcal CPS induced a dose-dependent release of cytokines ( P01375 -α , P05231 , P10145 , and P02778 ) and NO from human and murine macrophages , respectively . CPS induced P10145 release from P29320 cells stably transfected with O60603 /6 , O60603 , O60603 / P08571 , and O00206 /MD-2/ P08571 but not P29320 cells alone . mAb to O60603 but not an isotype control antibody blocked CPS-induced P10145 release from P29320 - O60603 /6-transfected cells . A significant reduction in P01375 -α and P10145 release was seen when THP-1- and P29320 - O00206 /MD-2- P08571 - but not P29320 - O60603 - or P29320 - O60603 /6-transfected cells were stimulated with CPS in the presence of DB04933 ( E5564 ) , a lipid A antagonist that binds to MD-2 , and a similar reduction in NO and P01375 -α release was also seen in RAW 264.7 cells in the presence of DB04933 . P08571 and P18428 enhanced CPS bioactivity , and NF-κB was , as anticipated , the major signaling pathway . Thus , these data suggest that innate immune recognition of meningococcal CPS by macrophages can occur via O60603 - and O00206 -MD-2 pathways . DB09036 for multicentric Castleman disease . Dysregulated secretion of P05231 plays a pivotal role in the pathogenesis of Castleman disease ( CD ) , a rare lymphoproliferative disorder . In contrast to unicentric CD for which surgery is considered the treatment of choice , there is no standard therapeutic approach for multicentric CD ( O95822 ) . DB09036 ( trade name : Sylvant , formerly known as CNTO 328 ) is a chimeric monoclonal antibody with high binding affinity for human P05231 . In a recent randomized placebo-controlled Phase II trial , subjects with HIV-negative , HHV8-negative O95822 who received siltuximab demonstrated a significantly higher rate of durable tumor and symptomatic response with a tolerable safety profile , leading to its approval for the treatment of HIV-negative HHV8-negative O95822 by the US FDA and the European Commission in April and May 2014 , respectively . This article will cover the current treatment options of O95822 , the drug profile of siltuximab and future directions in the management of O95822 . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Phase 1 study in Japan of siltuximab , an anti- P05231 monoclonal antibody , in relapsed/refractory multiple myeloma . DB09036 , a chimeric monoclonal antibody with high affinity and specificity for interleukin-6 , has been shown to enhance anti-multiple myeloma activity of bortezomib and corticosteroid in vitro . We evaluated the safety , pharmacokinetics , immunogenicity , and antitumor effect of siltuximab in combination with bortezomib and dexamethasone in Japanese patients with relapsed or refractory multiple myeloma . This open-label , phase 1 , dose-escalating study used two doses of siltuximab : 5.5 and 11.0 mg/kg ( administered on day 1 of each 21-day cycle ) . In total , nine patients were treated . The most common grade 3/4 adverse events , lymphopenia ( 89 % ) and thrombocytopenia ( 44 % ) , occurred in patients receiving both doses of siltuximab ; however , no dose-limiting toxicities ( DLTs ) were observed . Following intravenous administration of siltuximab at 5.5 and 11.0 mg/kg , the maximum serum concentration and the area under the curve from 0 to 21 days and from 0 to infinity increased in an approximately dose-proportional manner . Mean half-life , total systemic clearance , and volume of distribution were similar at doses of 5.5 and 11.0 mg/kg . Across both doses , six of the nine patients had complete or partial response ( 22 and 44 % , respectively ) . In conclusion , as no DLT was observed , the recommended dose for this combination is 11.0 mg/kg once every 3 weeks . The study is registered at http://www.clinicaltrials.gov as NCT01309412 . Temporal network based analysis of cell specific vein graft transcriptome defines key pathways and hub genes in implantation injury . Vein graft failure occurs between 1 and 6 months after implantation due to obstructive intimal hyperplasia , related in part to implantation injury . The cell-specific and temporal response of the transcriptome to vein graft implantation injury was determined by transcriptional profiling of laser capture microdissected endothelial cells ( EC ) and medial smooth muscle cells ( SMC ) from canine vein grafts , 2 hours ( H ) to 30 days ( D ) following surgery . Our results demonstrate a robust genomic response beginning at 2 H , peaking at 12-24 H , declining by 7 D , and resolving by 30 D . Gene ontology and pathway analyses of differentially expressed genes indicated that implantation injury affects inflammatory and immune responses , apoptosis , mitosis , and extracellular matrix reorganization in both cell types . Through backpropagation an integrated network was built , starting with genes differentially expressed at 30 D , followed by adding upstream interactive genes from each prior time-point . This identified significant enrichment of P05231 , P10145 , NF-κB , dendritic cell maturation , glucocorticoid receptor , and Triggering Receptor Expressed on Myeloid Cells ( Q9NP99 ) signaling , as well as PPARα activation pathways in graft EC and SMC . Interactive network-based analyses identified P05231 , P10145 , IL-1α , and P01308 Receptor ( P06213 ) as focus hub genes within these pathways . Real-time PCR was used for the validation of two of these genes : P05231 and P10145 , in addition to Collagen 11A1 ( P12107 ) , a cornerstone of the backpropagation . In conclusion , these results establish causality relationships clarifying the pathogenesis of vein graft implantation injury , and identifying novel targets for its prevention . DB09036 : first global approval . The anti-interleukin-6 ( P05231 ) chimeric monoclonal antibody siltuximab is the first drug to be approved for the treatment of multicentric Castleman 's disease ( O95822 ) in the US and European union ( EU ) , having gained approval under the FDA priority review program in the US and from an accelerated assessment and recommendation by the Committee for Medicinal Products for Human Use ( CHMP ) in the EU . Development of the drug is continuing in smoldering multiple myeloma . This article summarizes the milestones in the development of siltuximab leading to this first approval for O95822 . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . P40933 affects serotonin system and exerts antidepressive effects through IL15Rα receptor . Contrary to the reduction of depressive-like behavior observed in several strains of cytokine receptor knockout mice , mice lacking the specific receptor for interleukin ( IL ) -15 showed increased immobility in tail suspension and modified forced swimming tests . There was also a reduction in social interactions . The hippocampus of the IL15Rα knockout mice had decreased mRNA for 5-HT(1A) , increased mRNA for 5-HT(2C) , and region-specific changes of serotonin reuptake transporter ( P31645 ) immunoreactivity . DB00472 ( the classic antidepressant DB00472 , which inhibits 5-HT(2C) and P31645 ) reduced the immobility of the IL15Rα knockout mice in comparison with their pretreatment baseline . Together with the unchanged performance of the IL15Rα knockout mice on the rotarod , this response to fluoxetine indicates that the immobility reflects depression . Wildtype mice responded to P40933 treatment with improvement of immobility induced by forced swimming , whereas the knockout mice failed to respond . Thus , the cognate P40933 receptor is necessary for the antidepressive activity of P40933 . In ex vivo studies , P40933 decreased synaptosomal uptake of 5-HT , and modulated the expression of 5-HT(2C) and P31645 in cultured neurons in a dose- and time-dependent manner . Thus , the effect of P40933 on serotonin transmission may underlie the depressive-like behavior of IL15Rα knockout mice . We speculate that P40933 is essential to maintain neurochemical homeostasis and thereby plays a role in preventing neuropsychiatric symptoms . Randomised phase II study of siltuximab ( CNTO 328 ) , an anti- P05231 monoclonal antibody , in combination with mitoxantrone/prednisone versus mitoxantrone/prednisone alone in metastatic castration-resistant prostate cancer . PURPOSE : This open-label phase II trial assessed mitoxantrone/prednisone ( M/P ) with and without siltuximab ( CNTO 328 ) , an anti-interleukin-6 chimeric monoclonal antibody , for patients with metastatic castration-resistant prostate cancer who received prior docetaxel-based chemotherapy . METHODS : Part 1 assessed the safety of biweekly siltuximab 6 mg/kg plus M 12 mg/m(2) every 3 weeks and P. Part 2 assessed efficacy and safety of siltuximab plus M/P versus M/P alone . The primary end-point was progression-free survival ( PFS ) . Progression was defined as progressive disease per Response Evaluation Criteria in Solid Tumours ( RECIST ) , or ≥ 3 new skeletal lesions with clinical deterioration or without deterioration confirmed by repeated bone scan . Rising prostate-specific antigen was not considered progression . RESULTS : DB09036 plus M/P was well tolerated in Part 1 ( n=9 ) . In Part 2 , 48 and 49 patients received siltuximab plus M/P or M/P alone , respectively . Enrolment was prematurely terminated by the Independent Data Monitoring Committee since an apparent imbalance in patient baseline characteristics ( favoring the M/P only arm ) made it unlikely that the study could achieve its primary efficacy end-point . Median PFS was 97 days with siltuximab combination and 228 days with M/P alone ( hazard ratio , 1.72 ; P=0.043 ) . Use of a novel non-validated PFS definition may have contributed to this result . Abnormal laboratory assessments were more frequent with the combination . Infection and febrile neutropenia rates were similar between groups . Greater P02741 suppression was achieved during siltuximab combination treatment compared with M/P alone ( P=0.0003 ) . CONCLUSION : While siltuximab plus M/P appeared well tolerated , improvement in outcomes was not demonstrated . Antitumor efficacy of the anti-interleukin-6 ( P05231 ) antibody siltuximab in mouse xenograft models of lung cancer . INTRODUCTION : P05231 ( P05231 ) can activate downstream signaling pathways in lung cancer cells , such as the P40763 pathway , and is reported to be produced by tumor cells with activating P00533 mutations . We examined P05231 / P40763 in lung cancer tumor tissues and the effects of siltuximab , a neutralizing antibody to human P05231 , in mouse models of lung cancer . METHODS : P05231 and P40763 activation levels were compared with tumor histology and presence of P01116 mutations in snap-frozen , non-small-cell lung cancer tumors . The effects of siltuximab alone or in combination with erlotinib were examined in mouse xenograft models constructed using three cell line xenograft models and one primary explant mouse model . We examined the influence of cancer-associated fibroblasts ( CAFs ) on tumor growth and siltuximab effects . RESULTS : P05231 levels were higher in tumors of squamous cell versus adenocarcinoma histology and were not associated with presence of P01116 mutations . Tyrosine phosphorylation status of P40763 did not correlate with tumor P05231 levels . DB00133 phosphorylation of P40763 was correlated with P01116 mutation status . Both tumor and stromal cells contributed to total P05231 within tumors . DB09036 had minimal effect as a single agent in xenografts with tumor cells alone ; however , in models coadministered with CAFs , siltuximab had more potent effects on tumor inhibition . We observed no effects of combined erlotinib and siltuximab . CONCLUSIONS : P05231 is elevated in subsets of human NSCLCs , especially with squamous cell histology . Tumors supported by stromal production of P05231 seem to be the most vulnerable to tumor growth inhibition by siltuximab . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . High dose antithrombin III inhibits P09429 and improves endotoxin-induced acute lung injury in rats . OBJECTIVE : High mobility group box 1 ( P09429 ) is an important factor in the development of sepsis . Previous work suggests that antithrombin III ( P01008 ) inhibits inflammation , but the mechanism of action is still poorly understood . DESIGN AND SETTING : Prospective controlled animal study in a university laboratory . MATERIALS : Rats were randomly divided into a lipopolysaccharide ( LPS ) -induced sepsis control group and an P01008 -treated experimental group . Animals in the experimental group received a bolus of 250 units/kg of P01008 injected into the tail vein . MEASUREMENTS AND RESULTS : Animals receiving high-dose P01008 ( 250 units/kg ) had significantly improved lung histopathology and survival compared to the control rats . We measured serum and lung levels of various cytokines and P09429 at regular intervals from 0 to 12 h after the induction of sepsis and demonstrated lower P09429 levels over time in P01008 -treated animals . In an in vitro experiment , we stimulated the mouse macrophage cell line RAW 264.7 with LPS in the presence or absence of P01008 . Subsequent measurement of P09429 concentrations in the supernatant and cell signaling molecules in cell lysates revealed an P01008 dose-dependent decrease in P09429 release . Furthermore , inhibition of IkB and Q8NFH3 phosphorylation was observed with the administration of P01008 , suggestive of downstream signaling pathways . CONCLUSIONS : High-dose P01008 decreases lung pathology and reduces mortality in a rat sepsis model . This finding may be mediated by the inhibition of P09429 . P28335 receptor involvement in female rat lordosis behavior . Adult , hormone-primed , ovariectomized rats ( P05231 -344 ) with bilateral implants within the ventromedial nucleus of the hypothalamus ( VMN ) , were injected with 0.5 microgram estradiol benzoate followed 48 h later with 500 microgram progesterone . This priming produced rats with 2 different levels of sexual receptivity . Rats with a lordosis to mount ratio ( L/M ) >/= 0.5 were used to examine the potential lordosis-inhibiting effects of the 5- Q13049 receptor antagonist , R(+)-a- ( 2 , 3-dimethoxyphenyl ) -1-[2(4-fluoro-phenylethyl)]-4-piperidine-methanol ( MDL 100,907 ) , and the P28335 receptor antagonist , 5-methyl-1-(3-pyridylcarbamoyl)-1,2,3,5-tetrahydropyrrolo [ 2 , 3-f ] indole ( SB 206553 ) . Rats with low sexual receptivity ( L/M < 0.5 ) were bilaterally infused with the 5- Q13049 /2C receptor agonist , (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl ( DOI ) , or DOI plus either MDL 100,907 or SB 206553 to determine if either drug would attenuate the lordosis-facilitating effects of DOI . The P28335 receptor antagonist , but not the 5- Q13049 receptor antagonist , effectively inhibited lordosis behavior . Similarly , SB 206553 was more effective than MDL 100,907 in reducing the DOI-induced increase in lordosis responding . However , both drugs limited the duration of lordosis responding initiated by DOI . These results are consistent with prior suggestions that 5- Q13049 /2C receptors within the VMN are involved in the modulation of lordosis behavior and lead to the suggestion that P28335 , rather than 5- Q13049 , receptors are primarily responsible for the effects of 5-HT2 receptor-active drugs on lordosis behavior . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE .

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Regulation of P40763 by histone deacetylase-3 in diffuse large B-cell lymphoma : implications for therapy . Diffuse large B-cell lymphoma ( DLBCL ) with an activated B-cell ( DB01048 ) gene-expression profile has been shown to have a poorer prognosis compared with tumors with a germinal center B-cell type . ABC cell lines have constitutive activation of P40763 ; however , the mechanisms regulating P40763 signaling in lymphoma are unknown . In studies of class-I histone deacetylase ( HDAC ) expression , we found overexpression of O15379 in phospho P40763 -positive DLBCL and the O15379 was found to be complexed with P40763 . Inhibition of HDAC activity by DB06603 ( LBH589 ) increased p300-mediated P40763 (Lys685) acetylation with increased nuclear export of P40763 to the cytoplasm . HDAC inhibition abolished P40763 (Tyr705) phosphorylation with minimal effect on P40763 (Ser727) and O60674 tyrosine activity . pSTAT3(Tyr705)-positive DLBCLs were more sensitive to HDAC inhibition with LBH589 compared with pSTAT3(Tyr705)-negative DLBCLs . This cytotoxicity was associated with downregulation of the direct P40763 target Mcl-1 . O15379 knockdown upregulated P40763 (Lys685) acetylation but prevented P40763 (Tyr705) phosphorylation and inhibited survival of pSTAT3-positive DLBCL cells . These studies provide the rationale for targeting P40763 -positive DLBCL tumors with HDAC inhibitors . DB00169 upregulated protein 1 suppresses P01375 -α-induced NF-κB activation in hepatocarcinogenesis . Vitamin D(3) upregulated protein 1 ( Q9H3M7 ) is a candidate tumor suppressor , the expression of which is dramatically reduced in various tumor tissues . In this study , we found that Q9H3M7 expression is suppressed during human hepatic carcinogenesis , and mice lacking Q9H3M7 are much more susceptible to diethylnitrosamine-induced hepatocarcinogenesis compared with wild type mice . Q9H3M7 -deficient tumors proliferated significantly more than wild type tumors and had corresponding changes in the expression of key cell cycle regulatory proteins . In addition , the hepatomitogen-induced response was associated with a considerable increase in the release of P01375 -α and subsequent enhancement of NF-κB activation in Q9H3M7 -deficient mice . When cells were treated with P01375 -α , the Q9H3M7 level was markedly reduced , concomitant with elevated NF-κB activation . Furthermore , the overexpression of Q9H3M7 resulted in the robust suppression of P01375 -α-activated NF-κB activity via association with Q13547 and O15379 . These results indicate that Q9H3M7 negatively regulates hepatocarcinogenesis by suppressing P01375 -α-induced NF-κB activation . Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 , P05231 , and P13500 by Cbl-b(-/-) mast cells compared with levels produced by c-Cbl(-/-) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions . Treatment with DB06603 induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells . Increased levels of misfolded polypeptides in the endoplasmic reticulum ( ER ) triggers the dissociation of glucose-regulated protein 78 ( P11021 ) from the three transmembrane ER-stress mediators , i.e. , protein kinase RNA-like ER kinase ( Q9NZJ5 ) , activating transcription factor-6 ( P18850 ) , and inositol-requiring enzyme 1alpha , which results in the adaptive unfolded protein response ( UPR ) . In the present studies , we determined that histone deacetylase-6 ( Q9UBN7 ) binds and deacetylates P11021 . Following treatment with the pan-histone deacetylase inhibitor DB06603 ( Novartis Pharmaceuticals ) , or knockdown of Q9UBN7 by short hairpin RNA , P11021 is acetylated in 11 lysine residues , which dissociates P11021 from Q9NZJ5 . This is associated with the activation of a lethal UPR in human breast cancer cells . Coimmunoprecipitation studies showed that binding of Q9UBN7 to P11021 requires the second catalytic and COOH-terminal BUZ domains of Q9UBN7 . Treatment with DB06603 increased the levels of phosphorylated-eukaryotic translation initiation factor ( p-eIF2alpha ) , P18848 , and CAAT/enhancer binding protein homologous protein ( P35638 ) . DB06603 treatment also increased the proapoptotic Q13323 , O43521 , Q07812 , and Q16611 levels , as well as increased the activity of caspase-7 . Knockdown of P11021 sensitized MCF-7 cells to bortezomib and DB06603 -induced UPR and cell death . These findings indicate that enforced acetylation and decreased binding of P11021 to Q9NZJ5 is mechanistically linked to DB06603 -induced UPR and cell death of breast cancer cells . Mol Cancer Ther ; 9(4) ; 942-52 . (c)2010 AACR . Relationship between human oral lichen planus and oral squamous cell carcinoma at a genomic level : a datamining study . The leader gene approach is a data mining method based on the systematic search for genes involved in a specific process and their ranking according to the number of interconnections with the other genes identified . The genes with the strongest interconnections are termed leader genes , since they may be supposed to play an important role in the process . The potential of malignant progression of OLP to oral squamous cell carcinoma ( OSCC ) is still not completely clear . In this study , the leader gene approach is applied to investigate the association between OLP and OSCC at a molecular level . Results were integrated with those obtained in an experimental analysis ( see paper 1 of this series ) . Genes involved in OLP and OSCC were identified by systematic queries to dedicated databases . Interconnections among identified genes were calculated and given a confidence value using STRING database . Leader genes were identified by clustering genes according to their interconnections . This theoretical analysis shows that OLP and OSCC share two leader genes : P04637 and P38936 , involved in the PI3K signalling events mediated by AKT pathway . This finding and those obtained in the experimental analysis suggest the possible involvement of some key genes/proteins P06239 , P42336 , O15392 , P04637 and P38936 in the malignant progression from OLP to OSCC . Moreover , these findings support the role of some molecular pathways , namely P60568 signalling events mediated by PI3K , PI3K signalling events mediated by AKT , and , possibly , Aurora A signalling in the association between OLP and OSCC . P49771 promotes myeloid dendritic cell differentiation of human hematopoietic progenitor cells : possible application for cancer immunotherapy . Current in vitro culture systems allow the generation of human dendritic progenitor cells ( CFU-DCs ) . The aim of this study was to assess the effect of P49771 ( FL ) on the proliferation of human peripheral blood-derived myeloid CFU-DCs and their differentiation into more committed precursor cells ( pDCs ) using in vitro culture systems . Immunomagnetically separated P28906 + cells were cultured in serum-free , as well as in serum-containing , liquid suspension cultures to investigate the expansion and/or proliferation/differentiation of CFU-DCs , pDCs , and more mature dendritic cells ( DCs ) . FACS-sorted P28906 +Flt3+/- cells were cultured in methylcellulose to assay hematopoietic progenitors , including CFU-DCs . In the clonal cell culture supplemented with granulocyte/macrophage ( GM ) colony-stimulating factor ( P04141 ) , interleukin-4 , and tumor necrosis factor alpha , the frequency of CFU-DCs was significantly higher in the P28906 +Flt3+ fraction than in the P28906 +Flt3- population , thus suggesting functional Flt3 expression on CFU-DCs . Serum-free suspension culture of P28906 + cells revealed the potent effect of FL on the expansion of CFU-DCs in synergy with GM- P04141 and thrombopoietin ( P07202 ) . In addition , FL strongly induced the maturation of CFU-DCs into functional CD1a+ pDCs in serum-containing liquid suspension culture . Moreover , these FL-generated pDCs showed remarkable potential to differentiate into mature DCs with surface Q01151 / P42081 expression , which induced a distinct allogeneic T-cell response . These results clearly demonstrate that FL supports not only the proliferation of early hematopoietic progenitor cells , but also the maturation process of committed precursor cells along with the DC-lineage differentiation . Therefore , it is possible to develop a more efficient DC-based cancer immunotherapy using this specific cytokine combination , GM- P04141 + P07202 +FL in vitro in the near future . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Insufficient p65 phosphorylation at S536 specifically contributes to the lack of NF-kappaB activation and transformation in resistant JB6 cells . NF-kappaB activation is required for P01375 -induced transformation of JB6 mouse epidermal cells . Deficient activation of p65 contributes to the lack of NF-kappaB activation in transformation-resistant ( P- ) cells . We hypothesized that the differential NF-kappaB activation involves differential p65 phosphorylation arising from enzyme activity differences . Here we show that P01375 induces greater P29323 -dependent p65 phosphorylation at S536 in transformation sensitive ( P+ ) cells than in P- cells . Our results establish that limited P29323 content contributes to a low O15111 ( IKKbeta ) level , in turn resulting in insufficient p65 phosphorylation at S536 upon P01375 stimulation in P- cells . Phosphorylation of p65 at S536 appears to play a role in P01375 -induced p65 DNA binding and recruitment of p300 to the p65 complex as well as in release of p65 bound to Q13547 and 3 . Blocking p65 phosphorylation at S536 , but not at S276 or S529 , abolishes p65 transactivational activity . Over-expression of p65 but not p65 phosphorylation mutant ( S536A ) in transformation-resistant P- cells renders these cells sensitive to P01375 -induced transformation . Over-expression of p65 phosphorylation mimics p65-S536D or p65-S536E in P- cells and also rescues the transformation response . These findings provide direct evidence that phosphorylation of p65 at S536 is required for P01375 -induced NF-kappaB activation in the JB6 transformation model . The lack of NF-kappaB activation seen in P- cells can be attributed to an insufficient level of p65 phosphorylation on S536 that arises from insufficient IKKbeta that in turn arises from insufficient P29323 . Thus , p65 phosphorylation at S536 offers a potential molecular target for cancer prevention . Role of acetylation and extracellular location of heat shock protein 90alpha in tumor cell invasion . Heat shock protein ( hsp ) 90 is an DB00171 -dependent molecular chaperone that maintains the active conformation of client oncoproteins in cancer cells . An isoform , hsp90alpha , promotes extracellular maturation of matrix metalloproteinase ( MMP ) -2 , involved in tumor invasion and metastasis . Knockdown of histone deacetylase ( HDAC ) 6 , which deacetylates lysine residues in hsp90 , induces reversible hyperacetylation and attenuates DB00171 binding and chaperone function of hsp90 . Here , using mass spectrometry , we identified seven lysine residues in hsp90alpha that are hyperacetylated after treatment of eukaryotic cells with a pan-HDAC inhibitor that also inhibits Q9UBN7 . Depending on the specific lysine residue in the middle domain involved , although acetylation affects DB00171 , cochaperone , and client protein binding to hsp90alpha , acetylation of all seven lysines increased the binding of hsp90alpha to 17-allyl-amino-demethoxy geldanamycin . Notably , after treatment with the pan-HDAC inhibitor DB06603 ( LBH589 ) , the extracellular hsp90alpha was hyperacetylated and it bound to P08253 , which was associated with increased in vitro tumor cell invasiveness . Treatment with antiacetylated hsp90alpha antibody inhibited in vitro invasion by tumor cells . Thus , reversible hyperacetylation modulates the intracellular and extracellular chaperone function of hsp90 , and targeting extracellular hyperacetylated hsp90alpha may undermine tumor invasion and metastasis . Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene : interactive roles of modified histones , histone acetyltransferase , p300 , AND Sp1 . Atrial natriuretic peptide ( P01160 ) binds guanylyl cyclase-A/natriuretic peptide receptor-A ( P16066 /NPRA ) and produces the intracellular second messenger , cGMP , which regulates cardiovascular homeostasis . We sought to determine the function of histone deacetylases ( HDACs ) in regulating Npr1 ( coding for P16066 /NPRA ) gene transcription , using primary mouse mesangial cells treated with class-specific HDAC inhibitors ( HDACi ) . Trichostatin A , a pan inhibitor , and mocetinostat ( DB05651 ) , a class I HDAC inhibitor , significantly enhanced Npr1 promoter activity ( by 8- and 10-fold , respectively ) , mRNA levels ( 4- and 5.3-fold , respectively ) , and NPRA protein ( 2.7- and 3.5-fold , respectively ) . However , MC1568 ( class II HDAC inhibitor ) had no discernible effect . Overexpression of Q13547 and Q92769 significantly attenuated Npr1 promoter activity , whereas O15379 and Q9BY41 had no effect . HDACi-treated cultured cells in vitro and intact animals in vivo showed significantly reduced binding of Q13547 and -2 and increased accumulation of acetylated H3- P35527 /14 and H4-K12 at the Npr1 promoter . Deletional analyses of the Npr1 promoter along with ectopic overexpression and inhibition of Sp1 confirmed that HDACi-induced Npr1 gene transcription is accomplished by Sp1 activation . Furthermore , HDACi attenuated the interaction of Sp1 with Q13547 /2 and promoted Sp1 association with p300 and p300/ DB02527 -binding protein-associated factor ; it also promoted the recruitment of p300 and p300/ DB02527 -binding protein-associated factor to the Npr1 promoter . Our results demonstrate that trichostatin A and DB05651 enhanced Npr1 gene expression through inhibition of Q13547 /2 and increased both acetylation of histones ( H3- P35527 /14 , H4-K12 ) and Sp1 by p300 , and their recruitment to Npr1 promoter . Our findings define a novel epigenetic regulatory mechanism that governs Npr1 gene transcription . Combination therapy for hepatocellular carcinoma : additive preclinical efficacy of the HDAC inhibitor DB06603 with sorafenib . BACKGROUND & AIMS : Hepatocellular carcinoma ( HCC ) is a heterogeneous cancer in which sorafenib is the only approved systemic therapy . Histone deacetylases ( HDAC ) are commonly dysregulated in cancer and therefore represent promising targets for therapies , however their role in HCC pathogenesis is still unknown . We analyzed the expression of 11 HDACs in human HCCs and assessed the efficacy of the pan-HDAC inhibitor DB06603 alone and in combination with sorafenib in preclinical models of liver cancer . METHODS : Gene expression and copy number changes were analyzed in a cohort of 334 human HCCs , while the effects of DB06603 and sorafenib were evaluated in three liver cancer cell lines and a murine xenograft model . RESULTS : Aberrant HDAC expression was identified and validated in 91 and 243 HCCs , respectively . Upregulation of O15379 and Q9UQL6 mRNAs was significantly correlated with DNA copy number gains . Inhibiting HDACs with DB06603 led to strong anti-tumoral effects in vitro and vivo , enhanced by the addition of sorafenib . Cell viability and proliferation declined , while apoptosis and autophagy increased . DB06603 increased histone H3 and HSP90 acetylation , downregulated O15392 ( survivin ) and upregulated CDH1 . Combination therapy with DB06603 and sorafenib significantly decreased vessel density , and most significantly decreased tumor volume and increased survival in HCC xenografts . CONCLUSIONS : Aberrant expression of several HDACs and copy number gains of O15379 and Q9UQL6 occur in HCC . Treatment with DB06603 combined with sorafenib demonstrated the highest preclinical efficacy in HCC models , providing the rationale for clinical studies with this novel combination . c-Fes tyrosine kinase binds to and activates P40763 after granulocyte-macrophage colony-stimulating factor stimulation . Granulocyte-macrophage colony stimulating factor ( GM- P04141 ) induces proliferation and maturation of myeloid progenitor cells and also activates neutrophils . In order to investigate the pleiotropic effects of GM- P04141 stimulation , we examined the signaling pathways of protein tyrosine kinases ( PTKs ) and signal transducers and activators of transcription ( STATs ) in GM- P04141 -dependent proliferation of leukemia cells . Using TF-1 , a GM- P04141 -dependent human erythroleukemia cell line , we found that GM- P04141 enhanced DNA-binding and tyrosine phosphorylation of P40763 . GM- P04141 receptor ( GM-CSFR ) and c-Fes tyrosine kinase were also activated upon GM- P04141 stimulation . Furthermore , c-Fes formed a complex with P40763 . Experiments using a c-Fes mutant that lacked tyrosine kinase activity revealed that the activation of P40763 is kinase-dependent , but that the c-Fes- P40763 interaction is not affected by c-Fes tyrosine kinase activity . The results suggest that P40763 is activated by c-Fes tyrosine kinase through direct interaction during hematopoietic cell proliferation induced by GM- P04141 . R306465 is a novel potent inhibitor of class I histone deacetylases with broad-spectrum antitumoral activity against solid and haematological malignancies . R306465 is a novel hydroxamate-based histone deacetylase ( HDAC ) inhibitor with broad-spectrum antitumour activity against solid and haematological malignancies in preclinical models . R306465 was found to be a potent inhibitor of Q13547 and -8 ( class I ) in vitro . It rapidly induced histone 3 ( H3 ) acetylation and strongly upregulated expression of p21waf1,cip1 , a downstream component of Q13547 signalling , in A2780 ovarian carcinoma cells . R306465 showed class I HDAC isotype selectivity as evidenced by poor inhibition of Q9UBN7 ( class IIb ) confirmed by the absence of downregulation of Hsp90 chaperone c-raf protein expression and tubulin acetylation . This distinguished it from other HDAC inhibitors currently in clinical development that were either more potent towards Q9UBN7 ( e.g. vorinostat ) or had a broader HDAC inhibition spectrum ( e.g. DB06603 ) . R306465 potently inhibited cell proliferation of all main solid tumour indications , including ovarian , lung , colon , breast and prostate cancer cell lines , with IC50 values ranging from 30 to 300 nM . Haematological cell lines , including acute lymphoblastic leukaemia , acute myeloid leukaemia , chronic lymphoblastic leukaemia , chronic myeloid leukaemia , lymphoma and myeloma , were potently inhibited at a similar concentration range . R306465 induced apoptosis and inhibited angiogenesis in cell-based assays and had potent oral in vivo antitumoral activity in xenograft models . Once-daily oral administration of R306465 at well-tolerated doses inhibited the growth of A2780 ovarian , H460 lung and HCT116 colon carcinomas in immunodeficient mice . The high activity of R306465 in cell-based assays and in vivo after oral administration makes R306465 a promising novel antitumoral agent with potential applicability in a broad spectrum of human malignancies . Cyclophosphamide and other new agents for the treatment of severe aplastic anemia . Severe aplastic anemia ( P0DJI8 ) has a poor prognosis in the absence of treatment . Current accepted therapeutic strategies include allogeneic stem-cell transplantation and immunosuppression , both resulting in long-term survival in the majority of patients . Although human leukocyte antigen ( HLA ) -matched sibling stem-cell transplantation is highly effective , the 25 % probability of finding a suitable sibling donor within a family renders this approach available to only a minority of patients . Transplantation using HLA-matched , unrelated donors carries a high risk of treatment failure along with considerable toxicity . While combined immunosuppression with both antithymocyte globulin ( ATG ) and cyclosporine A ( Q13216 ) produces hematologic improvement in most patients , relapse is common . Late evolution of aplastic anemia to other serious hematologic disorders , including paroxysmal nocturnal hemoglobinuria ( PNH ) , myelodysplasia , and acute leukemia , is also a significant problem following treatment with ATG/ Q13216 . Recently , results of immunosuppression in P0DJI8 with another potent immunosuppressive agent , cyclophosphamide , were reported in a small number of patients . The overall response rate was similar to that seen with ATG/ Q13216 , but relapse and late clonal disease were not observed during a long period of follow-up . A larger randomized trial comparing sustained hematologic response rates to either conventional immunosuppression with ATG/ Q13216 or high-dose cyclophosphamide and Q13216 is now underway ; secondary end points include response duration , event-free survival , and overall survival . Additionally , a number of protocols designed to test the efficacy of alternative immunosuppressive or immunomodulatory agents are being developed . Kinin-B2 receptor exerted neuroprotection after diisopropylfluorophosphate-induced neuronal damage . The kinin-B2 receptor ( B2BKR ) activated by its endogenous ligand bradykinin participates in various metabolic processes including the control of arterial pressure and inflammation . Recently , functions for this receptor in brain development and protection against glutamate-provoked excitotoxicity have been proposed . Here , we report neuroprotective properties for bradykinin against organophosphate poisoning using acute hippocampal slices as an in vitro model . Following slice perfusion for 10min with diisopropylfluorophosphate ( DB00677 ) to initiate the noxious stimulus , responses of pyramidal neurons upon an electric impulse were reduced to less than 30 % of control amplitudes . Effects on synaptic-elicited population spikes were reverted when preparations had been exposed to bradykinin 30min after challenging with DB00677 . Accordingly , bradykinin-induced population spike recovery was abolished by HOE-140 , a B2BKR antagonist . However , the kinin-B1 receptor ( B1BKR ) agonist Lys-des- DB00125 (9)-bradykinin , inducing the phosphorylation of mitogen-activated protein kinase ( MEK/MAPK ) and cell death , abolished bradykinin-mediated neuroprotection , an effect , which was reverted by the P29323 inhibitor PD98059 . In agreement with pivotal B1BKR functions in this process , antagonism of endogenous B1BKR activity alone was enough for restoring population spike activity . On the other hand pralidoxime , an oxime , reactivating acetylcholinesterase ( P22303 ) after organophosphate poisoning , induced population spike recovery after DB00677 exposure in the presence of bradykinin and Lys-des- DB00125 (9)-bradykinin . Lys-des- DB00125 (9)-bradykinin did not revert protection exerted by pralidoxime , however when instead bradykinin and Ly-des- DB00125 (9)-bradykinin were superfused together , recovery of population spikes diminished . These findings again confirm the neuroprotective feature of bradykinin , which is , diminished by its endogenous metabolites , stimulating the B1BKR , providing a novel understanding of the physiological roles of these receptors . DB00188 prevents the expression of P45452 and the degradation of collagen type 2 in human chondrocytes . The structural backbone of extracellular matrix in cartilage is the collagen fibril , which is mainly composed of type II collagen . A measurable increase in type II collagen denaturation and degradation has been found in early Osteoarthritis ( OA ) . Pro-inflammatory cytokine such as P01375 -α produced in OA cartilage induced the expression of matrix metalloproteinase-13 ( P45452 ) , which targets and degrades type II collagen . DB00188 is a proteasome inhibitor approved by the FDA for treatment of multiple myeloma and mantel cell lymphoma . The effects of bortezomib in OA have not been reported before . In this study , we found that bortezomib is able to suppress the degradation of type II collagen induced by P01375 -α in human chondrocytes . Mechanistically , bortezomib treatment inhibits the expression of P10914 through blunting O60674 / P42224 pathway , thereby prevents the induction of P45452 as well as the degradation of type II collagen . Our findings suggest the therapeutic potentials of bortezomib in patients with OA . Comparison of the cytotoxicity of cladribine and clofarabine when combined with fludarabine and busulfan in AML cells : Enhancement of cytotoxicity with epigenetic modulators . Clofarabine ( Clo ) , fludarabine ( Flu ) , and busulfan ( Bu ) combinations are efficacious in hematopoietic stem cell transplantation for myeloid leukemia . We sought to determine whether the more affordable drug cladribine ( Clad ) can provide a viable alternative to Clo , with or without DB06603 ( Pano ) and DB01262 ( P22760 ) . Both Clad+Flu+Bu and Clo+Flu+Bu combinations showed synergistic cytotoxicity in KBM3/Bu250(6) , HL60 , and OCI- Q13950 cell lines . Cell exposure to these drug combinations resulted in 60 % -80 % inhibition of proliferation ; activation of the Q13315 pathway ; increase in histone modifications ; decrease in O15379 , P56524 , Q9UQL6 and SirT7 proteins ; decrease in mitochondrial membrane potential ; activation of apoptosis and stress signaling pathways ; and downregulation of the AKT pathway . These drug combinations activated DNA-damage response and apoptosis in primary cell samples from AML patients . At lower concentrations of Clad/Clo , Flu , and Bu , inclusion of Pano and P22760 enhanced cell killing , increased histone modifications and DNA demethylation , and increased the levels of P16/INK4a , P15/INK4b and P21/Waf1/Cip1 proteins . The observed DNA demethylating activity of Clad and Clo may complement P22760 activity ; increase demethylation of the gene promoters for Q8N474 , Q9UBP4 , and Q9Y5W5 ; and cause degradation of β-catenin in cells exposed to Clad/Clo+Flu+Bu+ P22760 +Pano . The overlapping activities of Clad/Clo+Flu+Bu , Pano , and P22760 in DNA-damage formation and repair , histone modifications , DNA demethylation , and apoptosis may underlie their synergism . Our results provide a basis for supplanting Clo with Clad and for including epigenetic modifiers in the pre-hematopoietic stem cell transplantation conditioning regimen for myeloid leukemia patients . [ Predictive factors of hormonal therapy in breast cancer ] . Breast cancer is a hormone-dependent cancer , and the presence of estrogen receptor ( ER ) and progesterone receptor ( PgR ) in tumors is used clinically to predict the likelihood of response to hormonal therapies . This review describes the roles of ( 1 ) hormone related factors ( ER , PgR , phosphorylated ER , ERbeta , aromatase ) , ( 2 ) growth related factors ( P04626 , Ki67 , p53 ) , ( 3 ) ER cofactors ( Q9Y6Q9 , NcoR1 ) , ( 4 ) estrogen dependent genes derived from gene expression profiling ( Q9UBN7 , P22692 /5 ) , and ( 5 ) gene profiling using cDNA microarray . There are , however , considerable methodological difficulties in identifying useful predictive factors but on the basis of current evidence other biomarkers add little additional information . The prospective and multi-centered analyses will be warranted to develop the predictive factors for directing use of these therapies . Dermoscopic , histological and immunohistochemical evaluation of cancerous features in acquired melanocytic nevi that have been repeatedly exposed to UVA or UVB . Previous studies have reported that repeated solar and artificial UVB ( 280-320 nm ) and UVA ( 320-400 nm ) exposures can modify acquired melanocytic nevi ( Q9BXJ7 ) . We therefore investigated the clinical , dermoscopic , histological and immunohistochemical changes in Q9BXJ7 exposed to UVB and UVA radiation . Twenty healthy volunteers with at least three Q9BXJ7 on the trunk were enrolled in the present study and randomized into two groups to receive equally effective doses of narrow-band ( NB ) -UVB or UVA1 . Three exposures per week were delivered for a total of 4 weeks . During exposures , one Q9BXJ7 was left unprotected , a second one was shielded with an opaque adhesive tape and the third nevus was covered with a commercial sunscreen . After the irradiation cycle , the Q9BXJ7 were surgically removed and underwent histological and immunohistochemical assessment of melanocyte/melanogenesis-related proteins ( Q16655 , tyrosinase , HMB-45 ) , cell cycle activation markers ( Ki-67 , topoisomerase IIalpha , p53 , Cdk2 ) and transcription factors ( microphthalmia-associated transcription factor , P40763 ) . Nevi that were exposed to NB-UVB or UVA1 also showed statistically significant increase in size and changes in their dermoscopic features , including overall darkening , increased pigment network expression , formation of branched streaks , and increased number and size of brown globules and dots . Q9BXJ7 that had been covered with opaque tape or sunscreen did not show changes in size or dermoscopic features following UVA1 or NB-UVB exposure . Histological and immunohistochemical analysis did not show any significant change in exposed Q9BXJ7 in comparison with Q9BXJ7 shielded with an opaque adhesive tape or covered with the sunscreen . DB06603 synergizes with bortezomib to induce endoplasmic reticulum stress and ubiquitinated protein accumulation in renal cancer cells . BACKGROUND : Inducing endoplasmic reticulum ( ER ) stress is a novel strategy used to treat malignancies . Inhibition of histone deacetylase ( HDAC ) 6 by the HDAC inhibitor DB06603 hinders the refolding of unfolded proteins by increasing the acetylation of heat shock protein 90 . We investigated whether combining DB06603 with the proteasome inhibitor bortezomib would kill cancer cells effectively by inhibiting the degradation of these unfolded proteins , thereby causing ubiquitinated proteins to accumulate and induce ER stress . METHODS : Caki-1 , ACHN , and 769-P cells were treated with DB06603 and/or bortezomib . Cell viability , clonogenicity , and induction of apoptosis were evaluated . The in vivo efficacy of the combination was evaluated using a murine subcutaneous xenograft model . The combination-induced ER stress and ubiquitinated protein accumulation were assessed . RESULTS : The combination of DB06603 and bortezomib induced apoptosis and inhibited renal cancer growth synergistically ( combination indexes < 1 ) . It also suppressed colony formation significantly ( p < 0.05 ) . In a murine subcutaneous tumor model , a 10-day treatment was well tolerated and inhibited tumor growth significantly ( p < 0.05 ) . Enhanced acetylation of the Q9UBN7 substrate alpha-tubulin was consistent with the suppression of Q9UBN7 activity by DB06603 , and the combination was shown to induce ER stress and ubiquitinated protein accumulation synergistically . CONCLUSIONS : DB06603 inhibits renal cancer growth by synergizing with bortezomib to induce ER stress and ubiquitinated protein accumulation . The current study provides a basis for testing the combination in patients with advanced renal cancer . Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors . The human HDAC ( histone deacetylase ) family , a well-validated anticancer target , plays a key role in the control of gene expression through regulation of transcription . While HDACs can be subdivided into three main classes , the class I , class II and class III HDACs ( sirtuins ) , it is presently unclear whether inhibiting multiple HDACs using pan-HDAC inhibitors , or targeting specific isoforms that show aberrant levels in tumours , will prove more effective as an anticancer strategy in the clinic . To address the above issues , we have tested a number of clinically relevant HDACis ( HDAC inhibitors ) against a panel of rhHDAC ( recombinant human HDAC ) isoforms . Eight rhHDACs were expressed using a baculoviral system , and a Fluor de Lystrade mark ( Biomol International ) HDAC assay was optimized for each purified isoform . The potency and selectivity of ten HDACs on class I isoforms ( rhHDAC1 , rhHDAC2 , rhHDAC3 and rhHDAC8 ) and class II HDAC isoforms ( rhHDAC4 , rhHDAC6 , rhHDAC7 and rhHDAC9 ) was determined . MS-275 was Q13547 -selective , DB05651 was Q13547 - and Q92769 -selective , apicidin was Q92769 - and O15379 -selective and valproic acid was a specific inhibitor of class I HDACs . The hydroxamic acid-derived compounds ( trichostatin A , DB00238 -LAQ824 , DB06603 , ITF2357 , vorinostat and belinostat ) were potent pan-HDAC inhibitors . The growth-inhibitory effect of the HDACis on HeLa cells showed that both pan-HDAC and class-I-specific inhibitors inhibited cell growth . The results also showed that both pan-HDAC and class-I-specific inhibitor treatment resulted in increased acetylation of histones , but only pan-HDAC inhibitor treatment resulted in increased tubulin acetylation , which is in agreement with their activity towards the Q9UBN7 isoform . Neisseria meningitidis capsular polysaccharides induce inflammatory responses via O60603 and O00206 -MD-2 . CPS are major virulence factors in infections caused by Neisseria meningitidis and form the basis for meningococcal serogroup designation and protective meningococcal vaccines . CPS polymers are anchored in the meningococcal outer membrane through a 1,2-diacylglycerol moiety , but the innate immunostimulatory activity of CPS is largely unexplored . Well-established human and murine macrophage cell lines and P29320 /TLR stably transfected cells were stimulated with CPS , purified from an endotoxin-deficient meningococcal serogroup B NMB-lpxA mutant . CPS induced inflammatory responses via O60603 - and O00206 -MD-2 . Meningococcal CPS induced a dose-dependent release of cytokines ( P01375 -α , P05231 , P10145 , and P02778 ) and NO from human and murine macrophages , respectively . CPS induced P10145 release from P29320 cells stably transfected with O60603 /6 , O60603 , O60603 / P08571 , and O00206 /MD-2/ P08571 but not P29320 cells alone . mAb to O60603 but not an isotype control antibody blocked CPS-induced P10145 release from P29320 - O60603 /6-transfected cells . A significant reduction in P01375 -α and P10145 release was seen when THP-1- and P29320 - O00206 /MD-2- P08571 - but not P29320 - O60603 - or P29320 - O60603 /6-transfected cells were stimulated with CPS in the presence of DB04933 ( E5564 ) , a lipid A antagonist that binds to MD-2 , and a similar reduction in NO and P01375 -α release was also seen in RAW 264.7 cells in the presence of DB04933 . P08571 and P18428 enhanced CPS bioactivity , and NF-κB was , as anticipated , the major signaling pathway . Thus , these data suggest that innate immune recognition of meningococcal CPS by macrophages can occur via O60603 - and O00206 -MD-2 pathways . A conserved structural motif reveals the essential transcriptional repression function of Spen proteins and their role in developmental signaling . Spen proteins regulate the expression of key transcriptional effectors in diverse signaling pathways . They are large proteins characterized by N-terminal RNA-binding motifs and a highly conserved C-terminal SPOC domain . The specific biological role of the SPOC domain ( Spen paralog and ortholog C-terminal domain ) , and hence , the common function of Spen proteins , has been unclear to date . The Spen protein , Q96T58 ( Q9Y618 / Q13547 -associated repressor protein ) , was identified as a component of transcriptional repression complexes in both nuclear receptor and Notch/ P02753 -Jkappa signaling pathways . We have determined the 1.8 A crystal structure of the SPOC domain from Q96T58 . This structure shows that essentially all of the conserved surface residues map to a positively charged patch . Structure-based mutational analysis indicates that this conserved region is responsible for the interaction between Q96T58 and the universal transcriptional corepressor Q9Y618 /NCoR ( silencing mediator for retinoid and thyroid receptors/nuclear receptor corepressor . We demonstrate that this interaction involves a highly conserved acidic motif at the C terminus of Q9Y618 /NCoR . These findings suggest that the conserved function of the SPOC domain is to mediate interaction with Q9Y618 /NCoR corepressors , and that Spen proteins play an essential role in the repression complex . The HDAC inhibitor LBH589 induces P29323 -dependent prometaphase arrest in prostate cancer via Q9UBN7 inactivation and down-regulation . Histone deacetylase inhibitors ( HDACIs ) have potent anti-cancer activity in a variety of cancer models . Understanding the molecular mechanisms involved in the therapeutic responsiveness of HDACI is needed before its clinical application . This study aimed to determine if a potent HDACI , LBH589 ( DB06603 ) , had differential therapeutic responsiveness towards LNCaP and PC-3 prostate cancer ( PCa ) cells . The former showed prometaphase arrest with subsequent apoptosis upon LBH589 treatment , while the latter was less sensitive and had late G2 arrest . The LBH589 treatment down-regulated Q9UBN7 and sustained P29323 activation , and contributed to prometaphase arrest . Mechanistically , LBH589 inhibited Q9UBN7 activity , caused its dissociation from protein phosphatase PP1α , and increased 14-3-3ζ acetylation . Acetylated 14-3-3ζ released its mask effect on serine 259 of c-Raf and serine 216 of Cdc25C subsequent to de-phosphorylation by PP1α , which contributed to P29323 activation . Enhanced P29323 activity by LBH589 further down-regulated Q9UBN7 protein levels and sustained P29323 activation by free-forward regulation . The sustained Cdc25C and P29323 activation resulted in early M-phase ( prometaphase ) arrest and subsequent apoptosis in the most sensitive LNCaP cells but not in PC-3 cells . This study provides pre-clinical evidence that Q9UBN7 may serve as a sensitive therapeutic target in the treatment of prostate cancer with HDACI LBH589 for clinical translation . This study also posits a novel mechanism of Q9UBN7 participation in regulating the c-Raf- P50391 - P29323 signaling pathway and contributing to M phase cell-cycle transition . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Pressure overload-induced cardiac hypertrophy response requires janus kinase 2-histone deacetylase 2 signaling . Pressure overload induces cardiac hypertrophy through activation of O60674 ( Jak2 ) , however , the underlying mechanisms remain largely unknown . In the current study , we tested whether histone deacetylase 2 ( Q92769 ) was involved in the process . We found that angiotensin II ( Ang-II ) -induced re-expression of fetal genes ( Atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) ) in cultured cardiomyocytes was prevented by the Jak2 inhibitor AG-490 and Q92769 inhibitor Trichostatin-A ( P32119 ) , or by Jak2/ Q92769 siRNA knockdown . On the other hand , myocardial cells with Jak2 or Q92769 over-expression were hyper-sensitive to Ang-II . In vivo , pressure overload by transverse aorta binding ( AB ) induced a significant cardiac hypertrophic response as well as re-expression of P01160 and DB04899 in mice heart , which were markedly reduced by AG-490 and P32119 . Significantly , AG-490 , the Jak2 inhibitor , largely suppressed pressure overload-/Ang-II-induced Q92769 nuclear exportation in vivo and in vitro . Meanwhile , P32119 or Q92769 siRNA knockdown reduced Ang-II-induced P01160 / DB04899 expression in Jak2 over-expressed H9c2 cardiomyocytes . Together , these results suggest that Q92769 might be a downstream effector of Jak2 to mediate cardiac hypertrophic response by pressure overload or Ang-II . Histone deacetylase inhibitor treatment dramatically reduces cholesterol accumulation in Niemann-Pick type C1 mutant human fibroblasts . Niemann-Pick type C ( NPC ) disease is predominantly caused by mutations in the O15118 protein that affect intracellular cholesterol trafficking and cause accumulation of unesterified cholesterol and other lipids in lysosomal storage organelles . We report the use of a series of small molecule histone deacetylase ( HDAC ) inhibitors in tissue culture models of NPC human fibroblasts . Some HDAC inhibitors lead to a dramatic correction in the NPC phenotype in cells with either one or two copies of the O15118 (I1061T) mutation , and for several of the inhibitors , correction is associated with increased expression of O15118 protein . Increased O15118 (I1061T) protein levels may partially account for the correction of the phenotype , because this mutant can promote cholesterol efflux if it is delivered to late endosomes and lysosomes . The HDAC inhibitor treatment is ineffective in an P61916 mutant human fibroblast line . Analysis of the isoform selectivity of the compounds used implicates Q13547 and/or Q92769 as likely targets for the observed correction , although other HDACs may also play a role . LBH589 ( DB06603 ) is an orally available HDAC inhibitor that crosses the blood-brain barrier and is currently in phase III clinical trials for several types of cancer . It restores cholesterol homeostasis in cultured O15118 mutant fibroblasts to almost normal levels within 72 h when used at 40 nM . The findings that HDAC inhibitors can correct cholesterol storage defects in human O15118 mutant cells provide the potential basis for treatment options for NPC disease . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Q9UBN7 sustains growth stimulation by prolonging the activation of P01133 receptor through the inhibition of rabaptin-5-mediated early endosome fusion in gastric cancer . The aberrant regulation of histone deacetylase 6 ( Q9UBN7 ) contributes to malignant progression in various types of cancer , but the mechanism underlying gastric carcinogenesis remains unknown . Aberrant Q9UBN7 overexpression was observed in a subset of human gastric cancer cells . Q9UBN7 knockdown caused the significant inhibition of gastric cancer cell growth without affecting the transition of cell cycles or the processing of cell death . We demonstrate that an increase in epidermal growth factor receptor ( P00533 ) signaling through decreased P00533 degradation was mediated by Q9UBN7 in gastric carcinogenesis . These results establish a molecular mechanism responsible for oncogenic Q9UBN7 , explaining how P00533 signaling induced by the growth factor is sustained during the malignant progression of gastric cancer . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . Autophagic impairment contributes to systemic inflammation-induced dopaminergic neuron loss in the midbrain . BACKGROUND : Neuroinflammation plays an important role in the pathogenesis of Parkinson 's disease ( PD ) , inducing and accelerating dopaminergic ( DA ) neuron loss . Autophagy , a critical mechanism for clearing misfolded or aggregated proteins such as α-synuclein ( α- O15061 ) , may affect DA neuron survival in the midbrain . However , whether autophagy contributes to neuroinflammation-induced toxicity in DA neurons remains unknown . RESULTS : Intraperitoneal injection of lipopolysaccharide ( LPS , 5 mg/kg ) into young ( 3-month-old ) and aged ( 16-month-old ) male C57BL/6J mice was observed to cause persistent neuroinflammation that was associated with a delayed and progressive loss of DA neurons and accumulation of α- O15061 in the midbrain . The autophagic substrate-p62 ( Q13501 ) persistently increased , whereas LC3-II and Q9UBN7 exhibited early increases followed by a decline . In vitro studies further demonstrated that P01375 -α induced cell death in PC12 cells . Moreover , a sublethal dose of P01375 -α ( 50 ng/ml ) increased the expression of LC3-II , p62 , and α- O15061 , implying that P01375 -α triggered autophagic impairment in cells . CONCLUSION : Neuroinflammation may cause autophagic impairment , which could in turn result in DA neuron degeneration in midbrain . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Deciphering the molecular and biologic processes that mediate histone deacetylase inhibitor-induced thrombocytopenia . Histone deacetylase inhibitor ( HDACI ) -induced thrombocytopenia ( DB01520 ) is a major dose-limiting toxicity of this new class of drugs . Using preclinical models to study the molecular and biologic events that underpin this effect of HDACI , we found that C57BL/6 mice treated with both the Q13547 /2-selective HDACI romidepsin and the pan-HDACI DB06603 developed significant DB01520 . HDACI-induced DB01520 was not due to myelosuppression or reduced platelet lifespan , but to decreased platelet release from megakaryocytes . Cultured primary murine megakaryocytes showed reductions in proplatelet extensions after HDACI exposure and a dose-dependent increase in the phosphorylation of myosin light chain 2 ( MLC2 ) . Phosphorylation of MLC to phospho-MLC ( pMLC ) and subsequent proplatelet formation in megakaryocytes is regulated by the Rho-GTPase proteins Rac1 , P60953 , and RhoA . Primary mouse megakaryocytes and the human megakaryoblastic cell line Meg-01 showed reductions in Rac1 , P60953 , and RhoA protein levels after treatment with HDACIs . We were able to overcome HDACI-induced DB01520 by administering the mouse-specific thrombopoietin ( P07202 ) mimetic AMP-4 , which improved platelet numbers to levels similar to untreated controls . Our report provides the first detailed account of the molecular and biologic processes involved in HDACI-mediated DB01520 . Moreover , our preclinical studies provide evidence that dose-limiting DB01520 induced by HDACIs may be circumvented using a P07202 mimetic . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Oxidative folding and assembly with transthyretin are sequential events in the biogenesis of retinol binding protein in the endoplasmic reticulum . DB00162 -binding protein ( P02753 ) is secreted out of the cell in its ligand-bound holo-form . The apo-form of P02753 is selectively retained within the endoplasmic reticulum ( ER ) by a mechanism that remains unknown . Using isolated microsomal system , we have recapitulated the biogenesis of P02753 involving its oxidative folding and assembly with transthyretin in the ER . In addition to dissecting its pathway of disulfide oxidation , we have analyzed association of its early folding intermediates with ER-chaperones . Our results show that of the three intramolecular disulfides present in P02753 ( 4-160 , 70-174 , and 120-129 ) the smallest loop ( 120-129 ) was most critical for P02753 to fold . Its absence caused P02753 to aggregate into an intermolecular disulfide-linked structure . After acquisition of the small loop , formation of one of the two big disulfides ( 4-160 or 70-174 ) was sufficient for P02753 to acquire a folded state . Using cross-linking in intact microsomes and sedimentation on sucrose gradients , we show that newly synthesized P02753 is associated with a complex of chaperones consisting of Grp94 , P11021 , P07237 , and calnexin . The complex was constitutively present in the ER , independent of the presence of folding substrates . P02753 dissociated from this complex coincident with the formation of one of the two big disulfide loops , whereas P02753 mutant lacking both the large disulfides showed persistent association . While highlighting the matrix-like characteristics of ER in isolated microsomal system our results provide insight into P02753 folding and assembly mechanisms that will aid our understanding of its complex secretion properties . Inflammatory protein serum amyloid A1 marks a subset of conventional renal cell carcinomas with fatal outcome . BACKGROUND : Conventional renal cell carcinoma ( RCC ) is the most common renal cancer . As the metastatic conventional RCC is practically incurable , there is a need for markers to estimate the tumour aggressiveness . OBJECTIVE : To identify and characterise new marker(s) associated with the poor prognosis of conventional RCC . DESIGN , SETTING , AND PARTICIPANTS : RNA from 24 conventional RCCs was analysed for global gene expression by Affymetrix U133 Plus 2.0 arrays . Tissue microarrays containing 224 renal tumours including 87 conventional RCCs were used for immunohistochemistry . Cell lines Q92769 , HD48 , HA344 and HA465 established in our laboratory were used for invasion assay and zymography . MEASUREMENTS : Serum amyloid A 1 ( P0DJI8 ) was found to be upregulated in conventional RCCs and it has been analysed by quantitative RT-PCR and immunohistochemistry on TMAs to establish the correlation between P0DJI8 protein expression and patient survival by uni and multivariate analysis . The effect of P0DJI8 on tumour cell behaviour in vitro has also been examined by invasion assay and zymography . RESULTS AND LIMITATIONS : P0DJI8 RNA is expressed in conventional RCC samples of patients with poor prognosis . Immunohistochemistry of 72 conventional RCCs with a 5 yr follow up showed a correlation between P0DJI8 expression and the clinical outcome of disease . Stimulation of conventional RCC cell lines with recombinant P0DJI8 increased the expression of metalloproteinase ( MMP ) -9 and the invasive potential of tumour cells . Limitation of the study is a relatively small number ( 72 ) of patients having follow up . CONCLUSION : P0DJI8 seems to be a useful marker to estimate the prognosis of conventional RCCs . Somatic mutations affect key pathways in lung adenocarcinoma . Determining the genetic basis of cancer requires comprehensive analyses of large collections of histopathologically well-classified primary tumours . Here we report the results of a collaborative study to discover somatic mutations in 188 human lung adenocarcinomas . DNA sequencing of 623 genes with known or potential relationships to cancer revealed more than 1,000 somatic mutations across the samples . Our analysis identified 26 genes that are mutated at significantly high frequencies and thus are probably involved in carcinogenesis . The frequently mutated genes include tyrosine kinases , among them the P00533 homologue Q15303 ; multiple ephrin receptor genes , notably P29320 ; vascular endothelial growth factor receptor P35968 ; and NTRK genes . These data provide evidence of somatic mutations in primary lung adenocarcinoma for several tumour suppressor genes involved in other cancers -- including P21359 , P25054 , P06400 and Q13315 -- and for sequence changes in P23468 as well as the frequently deleted gene Q9NZR2 . The observed mutational profiles correlate with clinical features , smoking status and DNA repair defects . These results are reinforced by data integration including single nucleotide polymorphism array and gene expression array . Our findings shed further light on several important signalling pathways involved in lung adenocarcinoma , and suggest new molecular targets for treatment . Early diagnosis of ataxia-telangiectasia using radiosensitivity testing . OBJECTIVES : To utilize radiosensitivity testing to improve early diagnosis of patients with ataxia-telangiectasia ( A-T ) . STUDY DESIGN : We established normal ranges for the colony survival assay ( Q13216 ) by testing cells from 104 patients with typical A-T , 29 phenotypic normal patients , and 19 A-T heterozygotes . We also analyzed 61 samples from patients suspected of having A-T and 25 patients with related disorders to compare the Q13216 with other criteria in the diagnosis of A-T . RESULTS : When cells were irradiated with 1.0 Gy , the mean survival fraction ( microSF +/- 1 SD ) for patients with A-T was 13.1 % +/- 7.2 % compared with 50.1 % +/- 13.5 % for healthy control patients . These data served to define a diagnostic range for the Q13216 ( ie , < 21 % ) , a normal range ( > 36 % ) , and a nondiagnostic intermediate range of 21 % to 36 % . The mutations of patients with A-T with intermediate radiosensitivity tended to cluster around the functional domains of the Q13315 gene . CONCLUSIONS : The Q13216 is a useful adjunctive test for confirming an early clinical diagnosis of A-T . However , Q13216 is also abnormal in other chromosomal instability and immunodeficiency disorders . Modulation of cytokine release from human monocytes by drugs used in the therapy of inflammatory bowel diseases . BACKGROUND : Cytokines produced in the gut mucosa play an important role in the pathogenesis of inflammatory bowel diseases ( Q9UKU7 ) . To determine whether drugs used in the treatment of these diseases modulate cytokine synthesis , we investigated their effects on endotoxin-induced tumour necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -1 beta and P05231 release by elutriation-purified human monocytes in vitro . METHODS : Drugs tested were dexamethasone , DB00244 , sulphapyridine and zileuton ( a P09917 inhibitor ) . Monocytes were isolated and stimulated with endotoxin , and P01375 , IL-1 and P05231 levels were determined using an enzyme-linked immunosorbent assay . RESULTS : Monocyte stimulation with endotoxin resulted in an average P01375 release of 2464 +/- 64 pg/10(6) cells , IL-1 release of 616 +/- 47 pg/10(6) cells and P05231 release of 2259 +/- 148 pg/10(6) cells . Addition of dexamethasone resulted in a reduction of P01375 , IL-1 and P05231 release to below background levels . DB00891 significantly reduced P01375 and induced IL-1 release in a dose-dependent fashion , but had no significant effect on P05231 release . 5- DB00233 did not modulate P05231 synthesis , but significantly reduced IL-1 and enhanced P01375 synthesis . Zileuton reduced P01375 and P05231 release , but enhanced IL-1 release . CONCLUSION : We conclude that these anti-inflammatory drugs are able to modulate cytokine release by human monocytes . Further studies are needed to determine whether these effects are related to their therapeutic efficacy in Q9UKU7 . Human mitochondrial transcription factor A functions in both nuclei and mitochondria and regulates cancer cell growth . Mitochondrial transcription factor A ( Q00059 ) is one of the high mobility group protein family and is required for both transcription from and maintenance of mitochondrial genomes . However , the roles of Q00059 have not been extensively studied in cancer cells . Here , we firstly reported the nuclear localization of Q00059 . The proportion of nuclear-localized Q00059 varied among different cancer cells . Some Q00059 binds tightly to the nuclear chromatin . DNA microarray and chromatin immunoprecipitation assays showed that Q00059 can regulate the expression of nuclear genes . Overexpression of Q00059 enhanced the growth of cancer cell lines , whereas downregulation of Q00059 inhibited their growth by regulating Q00059 target genes , such as baculoviral IAP repeat-containing 5 ( O15392 ; also known as survivin ) . Knockdown of Q00059 expression induced P38936 -dependent P55008 cell cycle arrest . These results imply that Q00059 functions in both nuclei and mitochondria to promote cell growth . Mechanisms of interleukin-1beta-induced P39905 release from rat glioma cells . P39905 ( P39905 ) is highly expressed both in neurons and astrocytes in injured tissues . Astrocytes support neurons by releasing neurotrophic factors including P39905 . It has been reported that various agents including cytokines such as interleukin ( IL ) -1beta induce P39905 mRNA expression and the release in astrocytes . However , the mechanism behind the P39905 synthesis and release remains unclear . Herein , we investigated the mechanisms of the IL-1beta-induced P39905 release from rat P13671 glioma cells . IL-1beta time dependently stimulated P39905 release from P13671 cells . IL-1beta induced the phosphorylation of inhibitor kappa B ( IkappaB ) , p38 mitogen-activated protein ( Q96HU1 ) kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase , stress-activated protein kinase/c-Jun N-terminal kinase ( SAPK/JNK ) and signal transducer and activator of transcription ( P35610 ) 3 . The IL-1beta-stimulated levels of P39905 were suppressed by wedelolactone , an inhibitor of O15111 , SB203580 , an inhibitor of p38 Q96HU1 kinase , PD98059 , an inhibitor of Q02750 /2 or Janus family of tyrosine kinase ( JAK ) inhibitor I , an inhibitor of upstream kinase of P40763 . On the contrary , SP600125 , an inhibitor of SAPK/JNK , failed to reduce the IL-1beta-effect . These results strongly suggest that IL-1beta stimulates P39905 release through the pathways of IkappaB-nuclear factor kappa B , p38 Q96HU1 kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase and JAK- P40763 , but not through the SAPK/JNK pathway in glioma cells . CD8 T cell-specific downregulation of histone hyperacetylation and gene activation of the P05112 gene locus by ROG , repressor of GATA . Chromatin remodeling of type 2 cytokine gene loci occurs during differentiation of naive P01730 and CD8 T cells into type 2 helper ( Th2 ) and cytotoxic ( Tc2 ) T cells . P05112 production and histone hyperacetylation in P05112 -associated nucleosomes in developing Tc2 cells were significantly lower than those of Th2 cells ; however , cytokine production and histone hyperacetylation of P05113 and P35225 genes were equivalent . Developing Tc2 cells expressed lower P23771 levels and dramatically increased levels of repressor of GATA ( ROG ) . A ROG response element in the P35225 gene exon 4 displayed Tc2-specific binding of ROG , Q13547 , and Q92769 and exhibited repression of P05112 gene activation . Thus , ROG may confer CD8 T cell-specific repression of histone hyperacetylation and activation of the P05112 gene locus . Regulation of matrix metalloproteinases ( MMP ) -2/-9 expression in eosinophilic chronic rhinosinusitis -- cell culture by interleukin-5 and -13 ? BACKGROUND : Eosinophils are a prominent immunological feature of chronic rhinosinusitis ( CRS ) . Cytokines in the respiratory mucosa may be the key to upper airway pathophysiology . Matrix metalloproteinases ( MMP ) represent an entire group of DB01593 dependent endopeptidases with the potential to alter the extracellular matrix ( Q13201 ) . In this study epithelial cultures of CRS were treated with interleukin ( IL ) -5 or P35225 and subsequent levels of metalloproteinases were determined . MATERIALS AND METHODS : The cells for CRS culture were obtained from patients undergoing functional endoscopic sinus surgery . After 8-72 hours incubation with 0.2-0.4 ng/ml P05113 or 3-6 ng/ml P35225 , the expression of the P08253 and -9 in the CRS cultures was analysed . RESULTS : After 72 hours incubation with P05113 , the relative levels of P08253 showed no significant alteration in protein expression in comparison with the control groups . Incubation with P35225 revealed a statistically insignificant decrease of the relative P14780 expression in ECRS compared to the control group ( p > 0.1 ) . CONCLUSION : Alterations of P08253 and -9 expression may play a role in ECRS , but the association with P05113 and P35225 remains unclear . Molecular mechanisms for transcriptional regulation of human high-affinity IgE receptor beta-chain gene induced by GM- P04141 . The beta-chain of the high-affinity receptor for IgE ( FcepsilonRI ) plays an important role in regulating activation of FcepsilonRI-expressing cells such as mast cells in allergic reactions . We already reported that the transcription factor myeloid zinc finger ( P28698 ) 1 which formed a high m.w. complex including four and a half LIM-only protein (FHL)3 in the nucleus repressed human beta-chain gene expression through an element in the fourth intron . We also found that GM- P04141 induced expression of P28698 and nuclear translocation of Q13643 . We screened a human cDNA library and identified NFY which was reported to bind histone deacetylases ( HDACs ) as a constituent of the complex . The C-subunit of NFY was demonstrated to form a ternary complex with P28698 / Q13643 and interact with a beta-chain gene region including the element in the fourth intron . Q13547 and Q92769 were also shown to interact with the fourth intron region of the beta-chain gene . In a human mast cell line HMC-1 cultured with GM- P04141 , both beta-chain expression and acetylation of histones interacting with the fourth intron region of the beta-chain gene were decreased . Collectively , these results indicated that HDACs , which were recruited to the beta-chain gene through the element in the fourth intron by P28698 / Q13643 /NFY , repressed beta-chain gene transcription by deacetylation of histones in the presence of GM- P04141 . These mechanisms will be involved in not only the cell type-specific repression of beta-chain gene expression in differentiating hemopoietic cells but also the repression of beta-chain gene expression in the peripheral cells under specific circumstances . Biphasic recruitment of transcriptional repressors to the murine cytomegalovirus major immediate-early promoter during the course of infection in vivo . Our previous studies showed that establishment of murine cytomegalovirus ( MCMV ) latency in vivo is associated with repression of immediate-early gene expression , deacetylation of histones bound to the major immediate-early promoter ( MIEP ) , changes in patterns of methylation of histones , and recruitment of cellular repressors of transcription to the MIEP . Here , we have quantitatively analyzed the kinetics of changes in viral RNA expression , DNA copy number , and recruitment of repressors and activators of transcription to viral promoters during the course of infection . Our results show that changes in viral gene expression correlate with changes in recruitment of RNA polymerase and acetylated histones to viral promoters . Binding of the transcriptional repressors histone deacetylase type 2 ( Q92769 ) , O15379 , P25490 , Q06330 / P02753 -Jk , Q9UER7 , and CIR to the MIEP and HDACs to other promoters showed a biphasic pattern : some binding was detectable prior to activation of viral gene expression , then decreased with the onset of transcription and increased again as repression of viral gene expression occurred . Potential binding sites for Q06330 / P02753 -Jk and P25490 in the MIEP and for P25490 in the M100 promoter ( M100P ) were identified by in silico analysis . While recruitment of HDACs was not promoter specific , binding of Q06330 / P02753 -Jk and P25490 was restricted to promoters with their cognate sites . Our results suggest that sequences within viral promoters may contribute to establishment of latency through recruitment of transcriptional repressors to these genes . The observation that repressors are bound to the MIEP and other promoters immediately upon infection suggests that latency may be established in some cells very early in infection . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Poly(A)-poly(U) induces circulating colony-stimulating activity resulting from interactions between endogeneous interleukin 6 and serum components . The immunostimulant poly(A)-poly(U) induces a rapid enhancement of circulating colony-stimulating activity ( Q13216 ) in normal mice , culminating 2 h after i.v. injection . A dose of 200 micrograms per mouse is sufficient for a maximal effect . The colonies formed in response to sera from poly(A)-poly(U)-injected mice are mainly granulocytic with few macrophages . These sera are devoid of detectable interleukin 3 ( P08700 ) or granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) , but contain large amounts of interleukin 6 ( P05231 ) that are perfectly correlated with circulating Q13216 levels . Although , in our hands , P05231 alone induces no colony formation in the standard methylcellulose colony assay , it is nevertheless requisite for this biological activity because 1 ) monoclonal antibodies against P05231 strongly diminish colony formation in response to sera from poly(A)-poly(U)-injected mice , and 2 ) recombinant (r) P05231 induces colonies when tested in combination with low amounts of normal murine serum . At the concentrations used ( 0.3 % -2.5 % ) , the latter has no or a very slight effect alone . Low amounts of hematopoietic growth factors , that is , macrophage colony-stimulating factor ( P09603 ) , granulocyte colony-stimulating factor ( DB00099 ) , GM- P04141 , or P08700 that are almost ineffective in the absence of P05231 can replace normal serum . Taken together , these data suggest that circulating P05231 , induced by i.v. injection of poly(A)-poly(U) , promotes colony formation by interacting with serum components that might be identical with hematopoietic growth factors present in normal serum at subliminal concentrations . Finally , the involvement of lipopolysaccharide ( LPS ) in this phenomenon has been ruled out by the use of the low responder strain of mice ( C3H/HeJ ) that leads to similar results . Expression of cytosolic retinoid-binding protein genes in human skin biopsies and cultured keratinocytes and fibroblasts . Using reverse transcription coupled to polymerase chain reaction we have studied the mRNA expression of serum retinol-binding protein and cytosolic receptors for retinol and retinoic acid in skin biopsies , and in cultured epidermal keratinocytes and dermal fibroblasts . Transcripts for cellular retinol-binding protein ( P09455 ) I and cellular retinoic-acid-binding protein ( CRABP ) I were found in normal skin , keratinocytes , and fibroblasts . CRABP II transcripts were detected in skin and keratinocytes . A decreased mRNA expression of CRABP I and an increased mRNA expression of CRABP II were found in lesional psoriatic skin compared with uninvolved skin . mRNA transcripts for serum retinol-binding protein ( s- P02753 ) were detected in all tissues and cells . The biological importance of s- P02753 expression in keratinocytes and fibroblasts is not known , but hypothetically this protein may be involved in the intracellular shuttling of retinol and retinoic acid , or in the retransportation of cellular retinoids into the extracellular space . Histone deacetylase inhibitors as potential therapeutic approaches for chordoma : an immunohistochemical and functional analysis . Chordomas are rare malignancies of the axial skeleton . Therapy is mainly restricted to surgery . This study investigates histone deacetylase ( HDAC ) inhibitors as potential therapeutics for chordomas . Immunohistochemistry ( IHC ) was performed using the HDAC 1-6 antibodies on 50 chordoma samples ( 34 primary tumors , 16 recurrences ) from 44 patients ( 27 male , 17 female ) . Pan-HDAC-inhibitors DB02546 ( DB02546 ) , DB06603 ( LBH-589 ) , and Belinostat ( PXD101 ) were tested for their efficacy in the chordoma cell line MUG-Chor1 via Western blot , cell cycle analysis , caspase 3/7 activity ( MUG-Chor1 , UCh-1 ) , cleaved caspase-3 , and PARP cleavage . p-Values below 0.05 were considered significant . IHC was negative for Q13547 , positive for Q92769 in most ( n = 36 ; 72 % ) , and for HDACs 3-6 in all specimens available ( n = 43 ; 86 % ) . Q9UBN7 expression was strongest . DB02546 and LBH-589 , but not PXD101 caused a significant increase of G2/M phase cells and of cleaved caspase-3 ( p = 0.0003 , and p = 0.0014 after 72 h , respectively ) , and a peak of caspase 3/7 activity . PARP cleavage confirmed apoptosis . The presented chordoma series expressed HDACs 2-6 with strongest expression of Q9UBN7 . DB02546 and LBH-589 significantly increased apoptosis and changed cell cycle distribution in vitro . HDAC-inhibitors should be further evaluated as therapeutic options for chordoma . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Destabilization of P04626 transcripts by targeting 3' untranslated region messenger RNA associated Q15717 and histone deacetylase-6 . In addition to repressing P04626 promoter function , histone deacetylase ( HDAC ) inhibitors induce the accelerated decay of mature P04626 transcripts ; the mechanism mediating this transcript destabilization is unknown but depends on the 3' untranslated region ( UTR ) of P04626 mRNA . Using P04626 -overexpressing human breast cancer cells ( SKBR3 ) , the mRNA stability factor Q15717 was shown to support P04626 transcript integrity , bind and endogenously associate with a conserved U-rich element within the P04626 transcript 3' UTR , coimmunoprecipitate with RNA-associated HDAC activity , and colocalize with Q9UBN7 . Q9UBN7 also coimmunoprecipitates with Q15717 in an RNA-dependent manner and within 6 hours of exposure to a pan-HDAC inhibitor dose , that does not significantly alter cytosolic Q15717 levels or Q15717 binding to P04626 mRNA . Cellular P04626 transcript levels decline while remaining physically associated with Q9UBN7 . Knockdown of Q9UBN7 protein by small interfering RNA partially suppressed the P04626 transcript decay induced by either pan-HDAC or Q9UBN7 -selective enzymatic inhibitors . Three novel hydroxamates , ST71 , ST17 , and ST80 were synthesized and shown to inhibit Q9UBN7 with 14-fold to 31-fold greater selectivity over their binding and inhibition of Q13547 . Unlike more potent pan-HDAC inhibitors , these Q9UBN7 -selective inhibitors produced dose-dependent growth arrest of P04626 -overexpressing breast cancer cells by accelerating the decay of mature P04626 mRNA without repressing P04626 promoter function . In sum , these findings point to the therapeutic potential of Q15717 and Q9UBN7 -selective inhibitors , contrasting P04626 stability effects induced by Q9UBN7 enzymatic inhibition and Q9UBN7 protein knockdown , and show that P04626 transcript stability mechanisms include exploitable targets for the development of novel anticancer therapies . Transient overexpression of mitochondrial transcription factor A ( Q00059 ) is sufficient to stimulate mitochondrial DNA transcription , but not sufficient to increase mtDNA copy number in cultured cells . Mitochondrial transcription factor A ( Q00059 ) stimulates transcription from mitochondrial DNA ( mtDNA ) promoters in vitro and in organello . To investigate whether changes of Q00059 levels also modulate transcription and replication in situ , the protein was transiently overexpressed in cultured cells . Mitochondrial mRNAs were significantly elevated at early time points , when no expansion of the Q00059 pool was yet observed , but were decreased when Q00059 levels had doubled , resemb-ling in vitro results . P29320 cells contain about 35 molecules of Q00059 per mtDNA . High levels of Q00059 were not associated with increases of full-length mtDNA , but nucleic acid species sensitive to RNAse H increased . Stimulation of transcription was more evident when Q00059 was transiently overexpressed in cells pre-treated with ethidium bromide ( EBr ) having lowered mtDNA , Q00059 and mitochondrial transcript levels . EBr rapidly inhibited mtDNA transcription , while decay of mtDNA was delayed and preferentially slowly migrating , relaxed mtDNA species were depleted . In conclusion , we show that transcription of mtDNA is submaximal in cultured cells and that a subtle increase of Q00059 within the matrix is sufficient to stimulate mitochondrial transcription . Thus , this protein meets all criteria for being a key factor regulating mitochondrial transcription in vivo , but other factors are necessary for increasing mtDNA copy number , at least in cultured cells . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Separate endocytic pathways regulate P05113 receptor internalization and signaling . Eosinophils are critically dependent on P05113 for their activation , differentiation , survival , and augmentation of cytotoxic activity . We previously showed that the cytoplasmic domain of the hematopoietic receptor , betac , which is shared by P05113 , P08700 , and GM- P04141 , is directly ubiquitinated and degraded by the proteasomes in a O60674 -dependent manner . However , studies describing the spatial distribution , endocytic regulation , and trafficking of betac-sharing receptors in human eosinophils are currently lacking . Using deconvolution microscopy and biochemical methods , we clearly demonstrate that IL-5Rs reside in and are internalized by clathrin- and lipid raft-dependent endocytic pathways . Microscopy analyses in TF1 cells and human eosinophils revealed significant colocalization of betac , IL-5Ralpha , and Cy3-labeled P05113 with transferrin- ( clathrin ) and cholera toxin-B- ( lipid raft ) positive vesicles . Moreover , whereas internalized IL-5Rs were detected in both clathrin- and lipid raft-positive vesicles , biochemical data revealed that tyrosine phosphorylated , ubiquitinated , and proteasome-degraded IL-5Rs partitioned to the soluble , nonraft fractions ( clathrin-containing ) . Lastly , we show that optimal P05113 -induced signaling requires entry of activated IL-5Rs into the intracellular compartment , as coimmunoprecipitation of key signaling molecules with the IL-5R was completely blocked when either endocytic pathway was inhibited . These data provide the first evidence that IL-5Rs segregate and traffic into two distinct plasma membrane compartments , and they further establish that IL-5R endocytosis regulates signaling both positively and negatively . The unexpected effect of cyclosporin A on CD56+CD16- and CD56+CD16+ natural killer cell subpopulations . DB00091 ( Q13216 ) is commonly used to prevent graft-versus-host disease . The influence of Q13216 on T-cell function has been extensively investigated ; however , the effect of Q13216 on natural killer ( NK ) cells is less understood . NK cells were cultured with P60568 and P40933 with and without Q13216 for 1 week . Compared with controls , Q13216 -treated cultures showed fewer CD56(+)CD16(+) P55040 (+) NK cells and a reciprocal increase in CD56(+)CD16(-) P55040 (-) cells . These changes were due mainly to a reduced proliferation of the CD56(dim) NK-cell subpopulation and a relative resistance of CD56(bright) NK cells to Q13216 . Following coculture with K562 targets , Q13216 -exposed NK cells differed from controls and lacked Ca(2+) oscillations , nuclear factor of activated T cells ( NFAT ) dephosphorylation , and NFAT nuclear translocation . NK cells cultured in Q13216 retained cytotoxicity against K562 , Raji , and P55040 ligand-expressing lymphoblastoid cells . NK cells cultured in Q13216 showed increases in O14931 and reductions in O95944 and P26718 . Following IL-12 and Q14116 stimulation , Q13216 -treated NK cells showed more P01579 -producing cells . Using in vitro NK-cell differentiation , progenitor cells gave rise to more CD56(+) P55040 (-) NK cells in the presence of Q13216 than controls . Collectively , these studies show that Q13216 influences NK-cell function and phenotype , which may have important implications for graft-versus-leukemia effects . Effects of retinol binding protein-4 on vascular endothelial cells . The study was designed to investigate the effect of retinol binding protein ( P02753 ) -4 on the phosphatidylinositol 3-kinase ( PI3K ) and mitogen-activated protein kinase ( MAPK ) pathways , which mediate the effects of insulin in vascular endothelial cells . The effects of P02753 on nitric oxide ( NO ) and insulin-stimulated endothelin-1 ( ET-1 ) secretion and on phosphorylation ( p ) of Akt , endothelial NO synthetase ( P29474 ) , and extracellular signal-regulated kinase ( P29323 )1/2 were investigated in bovine vascular aortic endothelial cells ( BAECs ) . P02753 showed an acute vasodilatatory effect on aortic rings of rats within a few minutes . In BAECs , P02753 -treatment for 5min significantly increased NO production , but inhibited insulin-stimulated ET-1 secretion . P02753 -induced NO production was not inhibited by tetraacetoxymethylester ( BAPTA-AM ) , an intracellular calcium chelator , but was completely abolished by wortmannin , a PI3K inhibitor . P02753 significantly increased p-Akt and p- P29474 production , and significantly inhibited p- P27361 /2 production . Triciribine , an Akt inhibitor , and wortmannin significantly inhibited P02753 -induced p-Akt and p- P29474 production . Inhibition of Akt1 by small interfering RNA decreased p- P29474 production enhanced by P02753 in human umbilical vein endothelial cells . In conclusion , P02753 has a robust acute effect of enhancement of NO production via stimulation of part of the PI3K/Akt/ P29474 pathway and inhibition of P27361 /2 phosphorylation and insulin-induced ET-1 secretion , probably in the MAPK pathway , which results in vasodilatation . Regulation of MyD88 aggregation and the MyD88-dependent signaling pathway by sequestosome 1 and histone deacetylase 6 . MyD88 is an essential adaptor molecule for Toll-like receptors ( TLRs ) and interleukin ( IL ) -1 receptor . MyD88 is thought to be present as condensed forms or aggregated structures in the cytoplasm , although the reason has not yet been clear . Here , we show that endogenous MyD88 is present as small speckle-like condensed structures , formation of which depends on MyD88 dimerization . In addition , formation of large aggregated structures is related to cytoplasmic accumulation of sequestosome 1 ( Q13501 ; also known as p62 ) and histone deacetylase 6 ( Q9UBN7 ) , which are involved in accumulation of polyubiquitinated proteins . A gene knockdown study revealed that Q13501 and Q9UBN7 were required for MyD88 aggregation and exhibited a suppressive effect on TLR ligand-induced expression of P05231 and NOS2 in RAW264.7 cells . Q13501 and Q9UBN7 were partially involved in suppression of several O00206 -mediated signaling events , including activation of p38 and JNK , but they hardly affected degradation of IκBα ( inhibitor of nuclear factor κB ) . Biochemical induction of MyD88 oligomerization induced recruitment of Q13501 and Q9UBN7 to the MyD88- Q9Y4K3 signaling complex . Repression of Q13501 and Q9UBN7 enhanced formation of the MyD88- Q9Y4K3 complex and conversely decreased interaction of the ubiquitin-specific negative regulator Q9NQC7 with the complex . Furthermore , ubiquitin-binding regions on Q13501 and Q9UBN7 were essential for MyD88 aggregation but were not required for interaction with the MyD88 complex . Thus , our study reveals not only that Q13501 and Q9UBN7 are important determinants of aggregated localization of MyD88 but also that MyD88 activates a machinery of polyubiquitinated protein accumulation that has a modulatory effect on MyD88-dependent signal transduction .

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Transcriptional modulation of monoaminergic neurotransmission genes by the histone deacetylase inhibitor trichostatin A in neuroblastoma cells . Histone deacetylase inhibitors are promising anti-tumor agents partly due to their ability to disrupt the hypoxic signaling pathway in human malignancies . However , little is known about any effects of these drugs on the central nervous system . The aim of the present study was to analyze the effects of trichostatin A ( P32119 ) -- a broad-spectrum histone deacetylase inhibitor -- on the transcriptional regulation of several genes involved in dopamine- and serotonergic neurotransmission . To this end , short-term parallel cultures of SK-NF-I neuroblastoma cells were treated with P32119 either alone or in combination with hypoxia , and mRNA levels of dopamine receptor D3 ( P35462 ) and D4 ( P21917 ) , dopamine transporter ( Q01959 ) , dopamine hydroxylase ( P09172 ) , dopamine receptor regulating factor ( DRRF ) , catechol-O-methyltransferase ( P21964 ) , serotonin receptor 1A ( P08908 ) , monoamino oxidase A ( P21397 ) , serotonin transporter ( P31645 ) and tryptophan hydroxylase 2 ( Q8IWU9 ) were determined by quantitative PCR . We found that P32119 did not antagonize the hypoxia-induced activation of D3 and D4 dopamine receptor genes , implying that induction of these genes is not mediated directly by hypoxia inducible factor-1alpha . On the other hand , P32119 dramatically upregulated the expression of Q01959 and P31645 ( 45-fold and 15-fold , respectively ) , while transcript levels of P21397 and P21964 were significantly reduced ( by 70 % and by more than 90 % , respectively ) . Induction of Q01959 protein expression was detected by western blotting . These results suggest that inhibition of histone deacetylases might help restore presynaptic monoamine pools via suppression of catecholamine breakdown and facilitation of monoamine reuptake in neurons . Novel camptothecin analogues that circumvent Q9UNQ0 -associated drug resistance in human tumor cells . DB00762 ( 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin ; CPT-11 ) is a widely used potent antitumor drug that inhibits mammalian P11387 ( Topo I ) ; however , overexpression of Q9UNQ0 ( Q9UNQ0 / Q9UNQ0 / Q9UNQ0 ) can confer cancer cell resistance to SN-38 , the active form of CPT-11 . We have recently demonstrated that plasma membrane vesicles prepared from Q9UNQ0 -overexpressing PC-6/ Q8WUX1 -5H cells transported SN-38 and its glucuronide conjugate in an DB00171 -dependent manner ( Nakatomi et al. , Biochem Biophys Res Commun 2001;288:827-32 ) . In the present study , we have characterized a total of 14 new camptothecin ( CPT ) analogues with respect to both the inhibition of Topo I and the substrate specificity of Q9UNQ0 . All of the tested CPT analogues , which have different substitutions at positions 10 and 11 , strongly inhibited the Topo I activity in a cell-free system , as did SN-38 . Their antitumor activities in the SN-38-resistant PC-6/ Q8WUX1 -5H2 cell line greatly varied , however , being correlated with intracellular accumulation levels . We have examined DB00171 -dependent transport of those CPT analogues by using plasma membrane vesicles prepared from both PC-6/ Q8WUX1 -5H2 cells and Q9UNQ0 -transfected P29320 -293 cells . Based on the substrate specificity of Q9UNQ0 thus evaluated , it is strongly suggested that CPT analogues with high polarity are good substrates for Q9UNQ0 and are therefore effectively extruded from cancer cells . In this context , to circumvent Q9UNQ0 -associated drug resistance , low-polarity CPT analogues are considered to be potent lead compounds . The present study provides a practical approach to discover new CPT-based drugs for the chemotherapy of drug-resistant human cancer . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Attenuated P08908 receptor signaling in brains of suicide victims : involvement of adenylyl cyclase , phosphatidylinositol 3-kinase , Akt and mitogen-activated protein kinase . Positron emission tomography studies in major depression show reduced serotonin (5-HT)1A receptor antagonist-binding potentials in many brain regions including occipital cortex . The functional meaning of this observation in terms of signal transduction is unknown . We used postmortem brain samples from depressed suicide victims to examine the downstream effectors of P08908 receptor activation . The diagnosis was established by means of psychological autopsy using Diagnostic and Statistical Manual of Mental Disorders ( DSM ) III-R criteria . Measurements of [35S]GTPgammaS binding to Galphai/o in the occipital cortex of suicide victims and matched controls revealed a blunted response in suicide subjects and a decrease in the coupling of P08908 receptor to adenylyl cyclase . No significant group differences were detected in the expression levels of Galphai/o , Galphaq/11 or Galphas proteins , or in the activity of DB02527 -dependent protein kinase A . Studies of a parallel transduction pathway downstream from P08908 receptor activation demonstrated a decrease in the activity of phosphatidylinositol 3-kinase and its downstream effector Akt , as well as an increase in P60484 ( phosphatase and tensin homolog deleted on chromosome 10 ) , the phosphatase that hydrolyzes phosphatidylinositol 3,4,5-triphosphate . Finally , the activation of extracellular signal-regulated kinases 1 and 2 was attenuated in suicide victims . These data suggest that the alterations in agonist-stimulated P08908 receptor activation in depressed suicide victims are also manifest downstream from the associated G protein , affecting the activity of second messengers in two P08908 receptor transduction pathways that may have implications for cell survival . P35372 and P00533 contribute to skin pigmentation differences between Indigenous Americans and Europeans . Contemporary variation in skin pigmentation is the result of hundreds of thousands years of human evolution in new and changing environments . Previous studies have identified several genes involved in skin pigmentation differences among African , Asian , and European populations . However , none have examined skin pigmentation variation among Indigenous American populations , creating a critical gap in our understanding of skin pigmentation variation . This study investigates signatures of selection at 76 pigmentation candidate genes that may contribute to skin pigmentation differences between Indigenous Americans and Europeans . Analysis was performed on two samples of Indigenous Americans genotyped on genome-wide SNP arrays . Using four tests for natural selection -- locus-specific branch length ( LSBL ) , ratio of heterozygosities ( lnRH ) , Tajima 's D difference , and extended haplotype homozygosity ( EHH ) -- we identified 14 selection-nominated candidate genes ( SNCGs ) . SNPs in each of the SNCGs were tested for association with skin pigmentation in 515 admixed Indigenous American and European individuals from regions of the Americas with high ground-level ultraviolet radiation . In addition to Q71RS6 and Q9UMX9 , genes previously associated with European/non-European differences in skin pigmentation , P35372 and P00533 were associated with variation in skin pigmentation in New World populations for the first time . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . A vitamin A deficient diet enhances proinflammatory cytokine , P35372 , and HIV-1 expression in the HIV-1 transgenic rat . The HIV-1 ( HIV ) transgenic ( Tg ) rat develops several immune abnormalities in association with clinical impairments that are similar to what are seen with HIV infection in humans . In HIV infection , retinoids and opioids can have separate and potentially combined effects on the clinical course of HIV disease . In these studies , the effects of a vitamin A deficient diet on T cell proinflammatory cytokine and mu opioid receptor ( MOR ) expression were examined in the Tg and in wild-type ( WT ) rats . The effects of the diet on HIV gene expression were also analyzed in the Tg rats . Phytohemagglutinin-stimulated T cells from WT rats on the vitamin A diet and from Tg rats on either diet were more likely to either produce increased percentages of T cells expressing intracytoplasmic P01579 , secrete higher levels of P01375 , and express higher levels of MOR mRNA and surface MOR . Mitogen stimulation also increased Tg rat HIV env , tat , and nef mRNA expression with even higher env and nef mRNA produced in association with the vitamin A deficient diet . All together , these data suggest that a vitamin A deficient diet can result in cellular effects that increase T cell proinflammatory responses and HIV expression , which may alter the course of disease in the HIV Tg rat model . Molecular determinants of trastuzumab efficacy : What is their clinical relevance ? DB00072 -containing therapy is a standard of care for human epidermal growth factor receptor-2 ( P04626 ) -positive breast cancer . In pre-clinical models , a wide range of molecular mechanisms have been associated with reduced sensitivity to trastuzumab in vitro . These include expression of the truncated P04626 receptor fragment p95HER2 , activating mutation of the gene encoding the class 1A catalytic subunit of phosphatidylinositol 3-kinase ( P42336 ) , loss of phosphatase and tensin homolog ( P60484 ) , activation of other downstream signal transducers , prevention of cell cycle arrest , increased signaling through alternative ( HER or non-HER ) tyrosine kinase receptors , and resistance to antibody-dependent cellular cytotoxicity . However , the clinical significance of these mechanisms as determinants of trastuzumab efficacy in vivo has been unclear . Here , we review clinical studies of potential predictive biomarkers of trastuzumab efficacy in P04626 -positive breast cancer and consider whether evaluation of such markers might inform patient selection for therapy . We find that clinical evidence relating to potential predictive biomarkers is mostly limited to small , retrospective studies , many of which have yielded conflicting findings . Some trends are evident in the retrospective data and in biomarker analyses from randomized clinical trials , particularly relating to activation of the phosphatidylinositol 3-kinase pathway , but none is sufficiently strong to form a basis for patient selection . This may be explained by the fact that multiple mechanisms of action determine the clinical efficacy of trastuzumab . In the absence of novel , validated biomarkers of efficacy , trastuzumab eligibility should continue to be based on evaluation of P04626 status according to standard methods . TATA-driven transcriptional initiation and regulation of the rat serotonin P08908 receptor gene . The transcriptional initiation and regulation of the rat serotonin P08908 receptor gene were characterized . By three types of analyses , a single brain-specific site of transcriptional initiation was localized to -967 bp upstream of the translation initiation codon that is utilized both in hippocampus and in the rat raphe RN46A cell line . This major site of transcriptional initiation was located 58 bp downstream from a consensus TATA element , suggesting TATA-driven transcription of the rat P08908 receptor . To identify the promoter activity of the receptor gene , progressive 5' deletions of the -2,719/-117-bp fragment of the P08908 promoter linked to luciferase gene were transfected into P08908 -negative ( pituitary GH4C1 , Q9BTT4 myoblast , and P13671 glioma ) and P08908 -positive ( septal SN-48 and raphe RN46A ) cell lines . Enhancer regions were identified within a fragment between nucleotides -426 and -117 that selectively enhanced transcription in P08908 -positive cells . A nonselective enhancer/promoter that mediated expression in all cell lines was located upstream between -1,519 and -426 bp in a DNA segment containing consensus TATA , CCAAT , SP-1 , and AP-1 elements as well as a poly-GT26 dinucleotide repeat . Strong repression of transcription in all cell lines was conferred by the region upstream of -1,519 bp that contains a 152-bp DNA segment with > 80 % identity to RANTES , tumor necrosis factor-beta , and other immune system genes . Our results indicate that TATA-driven expression of the P08908 receptor is regulated by a novel proximal tissue-specific enhancer region , a nonselective promoter , and an upstream repressor region that is distinct from previously identified neuron-specific repressors . Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 ) -induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 -Px , MDA , NO and P35228 , and the activities of serum ALT and Q9NRA2 caused by DB00316 . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 ) , pro-inflammatory mediators ( IL-1β , P05112 , P05231 , P01375 -α , P35228 , Bax , HMGB-1 and P35354 ) , pro-inflammatory transcription factors ( NF-κB and AP-1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase-3 , caspase-9 , p-JNK , p-p38 and p- P29323 ) , and increased the protein expressions of Bcl-2 and Bcl-xL . Moreover , the gene expression of P22301 , and the proteins including LC3 , Q14457 and Atg5 induced by DB00316 were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 -induced liver damage in the future . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Stimulatory effects of 5HT1A receptor agonists on luteinizing hormone-releasing hormone release from cultured fetal rat hypothalamic cells : interactions with progesterone . Previous works have suggested an interactive stimulatory effect of progesterone ( P ) and serotonin ( 5-HT ) on luteinizing hormone release . The purpose of the present study was to determine whether 5-HT via P08908 receptors interacts with P in the process of luteinizing hormone-releasing hormone ( P01148 ) release . Using fetal hypothalamic neurons in primary cell cultures the first goal of this study was to determine the effects of P08908 receptor agonists on P01148 secretion . 8-Hydroxy-2 ( di-n-propylamino ) tetralin ( 8-OH-DPAT ) or ipsapirone ( 10(-5) M ) significantly stimulated P01148 release . Pharmacological studies have allowed to rule out the possible involvement of alpha 2- or beta-adrenoreceptors , or 5-HT uptake sites , in the stimulatory effect of 8-OH-DPAT on P01148 release , thus demonstrating the specific involvement of P08908 receptors in the stimulation of P01148 release . The second goal was to test the ability of P to stimulate P01148 release from fetal hypothalamic neurons . P ( 10(-6) M ) applied for 30 or 120 min significantly stimulated P01148 secretion . The maintenance of the stimulation of P01148 release by P after a cycloheximide treatment or by an impermeable analog of P , P-3-BSA , has suggested a nongenomic effect of P on P01148 release . The effects of a pretreatment of cells by P on 8-OH-DPAT-induced P01148 release were tested . While 10(-7) M P alone did not stimulate P01148 release , this concentration of steroid potentiated the P01148 response to 10(-5) M 8-OH-DPAT . These findings led to the conclusion that P acting at the level of the plasma membrane potentiates the stimulatory effect of P08908 receptor agonists on P01148 release . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Transcriptional activation of human mu-opioid receptor gene by insulin-like growth factor-I in neuronal cells is modulated by the transcription factor Q13127 . The human mu-opioid receptor gene ( P35372 ) promoter contains a DNA sequence binding the repressor element 1 silencing transcription factor ( Q13127 ) that is implicated in transcriptional repression . We investigated whether insulin-like growth factor I ( P05019 ) , which affects various aspects of neuronal induction and maturation , regulates P35372 transcription in neuronal cells in the context of the potential influence of Q13127 . A series of P35372 -luciferase promoter/reporter constructs were transfected into two neuronal cell models , neuroblastoma-derived SH-SY5Y cells and PC12 cells . In the former , endogenous levels of human mu-opioid receptor ( hMOPr ) mRNA were evaluated by real-time PCR . P05019 up-regulated P35372 transcription in : PC12 cells lacking Q13127 , in SH-SY5Y cells transfected with constructs deficient in the Q13127 DNA binding element , or when Q13127 was down-regulated in retinoic acid-differentiated cells . P05019 activates the signal transducer and activator of transcription-3 signaling pathway and this transcription factor , binding to the signal transducer and activator of transcription-1/3 DNA element located in the promoter , increases P35372 transcription . We propose that a reduction in Q13127 is a critical switch enabling P05019 to up-regulate hMOPr . These findings help clarify how hMOPr expression is regulated in neuronal cells . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Anti-stress effect of astragaloside IV in immobilized mice . ETHNOPHARMACOLOGICAL RELEVANCE : Astragaloside IV , a major component extracted from the roots of Astragalus membranaceus ( AM ) , possesses anti-inflammatory , anti-oxidative , anti-fibrotic , anti-infarction and immunoregulatory effects . To clarify anti-stress effect of AM , anxiolytic and anti-inflammatory effects of 80 % ethanol extract of AM and astragaloside IV were investigated in immobilization stress model . MATERIALS AND METHODS : The mice were orally administered with AM ( 50 , 200 , and 500 mg/kg ) , astragaloside IV ( 5 , 10 , and 20 mg/kg ) and buspirone , a positive drug , 1h before immobilization treated for 2h . For anxiolytic activity assay , EPM test was performed in mice . For anti-inflammatory activity assay , serum levels of corticosterone , P05231 and P01375 -α were measured using ELISA kits . RESULTS : AM extract and astragaloside IV increased dose-dependently time spent on open arms and open arm entries in the EPM test . Anxiolytic effects of AM extract ( 500 mg/kg ) and astragaloside IV ( 20 mg/kg ) were comparable to those of buspirone ( 1 mg/kg ) . Their anxiolytic effects were blocked by WAY-100635 ( 0.5 mg/kg , i.p. ) , a P08908 receptor antagonist ( p < 0.01 ) , but not by flumazenil ( 3 mg/kg , i.p. ) and bicuculline ( 0.5 mg/kg , i.p. ) , GABAA receptor antagonists . AM extract and astragaloside IV also reduced serum levels of corticosterone , P05231 and P01375 -α dose-dependently . CONCLUSIONS : AM , particularly astragaloside IV , may ameliorate immobilized stress-induced anxiety and inflammation . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Role of serotonin in the regulation of interferon-gamma production by human natural killer cells . Monocytes , recovered directly from peripheral blood by counter-current centrifugal elutriation ( CCE ) , were shown to provide two regulatory signals for induction of interferon-gamma ( P01579 ) in natural killer ( NK ) cells in response to interleukin-2 ( P60568 ) : an upregulating signal and an inhibitory signal . The inhibitory signal was time-dependent , irreversible , and operating on a pretranslational level , as indicated by the inability of enriched NK cells to accumulate P01579 mRNA in the presence of elutriated monocytes . Monocyte-induced inhibition of P01579 production was abrogated by the biogenic amine serotonin , acting via the 5-hydroxytryptamine , or serotonin ( P08908 ) , subset of serotonin receptors ( 5-HTR ) . Thereby , serotonin effectively promoted P01579 production in the presence of monocytes . We conclude that serotonergic P08908 receptors transduce signals that are required for NK cells to produce P01579 in response to P60568 . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . DB00072 has preferential activity against breast cancers driven by P04626 homodimers . In breast cancer cells with P04626 gene amplification , P04626 receptors exist on the cell surface as monomers , homodimers , and heterodimers with P00533 / P21860 . The therapeutic antibody trastuzumab , an approved therapy for P04626 (+) breast cancer , can not block ligand-induced P04626 heterodimers , suggesting it can not effectively inhibit P04626 signaling . Hence , P04626 oligomeric states may predict the odds of a clinical response to trastuzumab in P04626 -driven tumors . To test this hypothesis , we generated nontransformed human MCF10A mammary epithelial cells stably expressing a chimeric P04626 -FKBP molecule that could be conditionally induced to homodimerize by adding the FKBP ligand AP1510 , or instead induced to heterodimerize with P00533 or P21860 by adding the heterodimer ligands P01133 /TGFα or heregulin . AP1510 , P01133 , and heregulin each induced growth of MCF10A cells expressing P04626 -FKBP . DB00072 inhibited homodimer-mediated but not heterodimer-mediated cell growth . In contrast , the P04626 antibody pertuzumab , which blocks P04626 heterodimerization , inhibited growth induced by heregulin but not AP1510 . Lastly , the P04626 / P00533 tyrosine kinase inhibitor lapatinib blocked both homodimer- and heterodimer-induced growth . AP1510 triggered phosphorylation of Erk1/2 but not AKT , whereas trastuzumab inhibited AP1510-induced Erk1/2 phosphorylation and Shc- P04626 homodimer binding , but not TGFα-induced AKT phosphorylation . Consistent with these observations , high levels of P04626 homodimers correlated with longer time to progression following trastuzumab therapy in a cohort of patients with P04626 -overexpressing breast cancer . Together , our findings confirm the notion that P04626 oligomeric states regulate P04626 signaling , also arguing that trastuzumab sensitivity of homodimers may reflect their inability to activate the PI3K ( phosphoinositide 3-kinase ) /AKT pathway . A clinical implication of our results is that high levels of P04626 homodimers may predict a positive response to trastuzumab . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . P35372 -dependent and independent components in effects of tramadol . DB00193 is thought to induce analgesia via both opioid and non-opioid pathways , although the precise mechanisms remain to be elucidated . In this study , we investigated the roles of the mu-opioid receptor ( MOP ) in analgesic and rewarding effects of tramadol by using MOP knockout ( KO ) mice . DB00193 -induced antinociception , assessed by hot-plate and tail-flick tests , was significantly reduced in heterozygous and homozygous MOP-KO mice when compared with that in wild-type mice . Interestingly , however , tramadol retained its ability to induce significant antinociception in homozygous MOP-KO mice . The tramadol-induced antinociception remaining in homozygous MOP-KO mice was not significantly affected by methysergide , a serotonin receptor antagonist , but was partially blocked by yohimbine , an adrenaline alpha2 receptor antagonist , and both naloxone , a non-selective opioid receptor antagonist , and yohimbine . In addition , antinociceptive effects of an active tramadol metabolite M1 were abolished or remarkably reduced in MOP-KO mice . On the other hand , neither wild-type nor homozygous MOP-KO mice showed significant place preference for tramadol in a conditioned place preference test , although there were slight tendencies toward preference in wild-type mice and avoidance in homozygous MOP-KO mice . These results strongly support the idea suggested in the previous pharmacological studies that MOP and the adrenaline alpha2 receptor mediate most of the analgesic properties of tramadol . DB00644 agonists reduce the migratory and the invasive behavior of androgen-independent prostate cancer cells by interfering with the activity of P05019 . Androgen-independent prostate carcinoma is characterized by a high proliferation rate and by a strong metastatic behavior . We have previously shown that DB00644 agonists exert a direct and specific inhibitory action on the proliferation of androgen-independent prostate cancer cells ( DU 145 ) . These compounds mainly act by interfering with the mitogenic activity of growth factors , such as the insulin-like growth factor-I ( P05019 ) . The present experiments were performed to clarify whether DB00644 agonists might also affect the migratory and the invasive behavior of androgen-independent prostate cancer cells and to define their mechanism of action . First we showed that the DB00644 agonist DB00007 reduces the migration of DU 145 cells towards a chemoattractant and their ability to invade a reconstituted basement membrane . Experiments were then performed to clarify whether the DB00644 agonist might act by interfering with the pro-metastatic activity of P05019 . We found that , in androgen-independent prostate cancer cells , DB00007 : a ) interferes with the P05019 system ( receptor protein expression and tyrosine-phosphorylation ) ; b ) abrogates the P05019 -induced phosphorylation of Akt ( a kinase previously shown by us to mediate the pro-metastatic activity of P05019 in prostate cancer cells ) ; c ) counteracts the migration and the invasive activity of the cells stimulated by P05019 ; d ) abolishes the effects of P05019 on cell morphology , on actin cytoskeleton organization and on alphavbeta3 integrin expression/cellular localization . These data indicate that DB00644 agonists , in addition to their well known antiproliferative effect , can also exert a significant inhibitory activity on the migratory and invasive behavior of androgen-independent prostate cancer cells , expressing the P30968 . DB00644 agonists act by interfering with the pro-metastatic activity of the growth factor P05019 .

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Interstitial pneumonitis associated with infliximab therapy without methotrexate treatment . P01375 ( P01375 ) -α inhibitors are increasingly being used to treat rheumatoid arthritis . DB00065 ( P27352 ) is a P01375 -α inhibitor that is usually used in combination with methotrexate ( MTX ) . Interstitial lung disease ( ILD ) during combination therapy has been attributed to MTX rather than P27352 . However , P27352 -associated ILD without combination with MTX has recently been reported . We describe herein a case of severe ILD secondary to P27352 without MTX therapy . New disease modifying agents in adult rheumatoid arthritis . In recent years , new disease modifying agents including leflunomide and tumour necrosis factor ( P01375 ) antagonists have been used to treat patients with rheumatoid arthritis ( RA ) . Leflunomide prevents proliferation of activated lymphocytes by inhibiting dihydroorotate dehydrogenase , a critical step in de novo pyrimidine synthesis . Leflunomide has been shown to be as effective as sulfasalazine and methotrexate ( MTX ) in placebo-controlled trials . It also improves physical function , quality of life measures and retards radiographic progression . P01375 antagonists include infliximab and etanercept . DB00065 is a chimeric P01375 monoclonal antibody . Repeated infusions of low dose infliximab ( 1 mg/kg ) are ineffective if given alone . Addition of MTX to infliximab has been shown to prolong the duration of clinical response . DB00005 is a human P01375 receptor p75 Fc fusion protein . In active RA patients with suboptimal response to MTX , additional clinical benefit was obtained by the addition of infliximab or etanercept to MTX . The main side effect of etanercept is injection site reaction . However , the long-term effect of P01375 antagonists in the development of infection , malignancy and autoimmune disease remains unknown . An antibody of P01375 did not prevent thioacetamide-induced hepatotoxicity in rats . P01375 ( P01375 ) -α antibodies have been shown to reduce liver damage in different models . We investigated the effects of infliximab ( a P01375 -α antibody ) on liver damage in thioacetamide ( TAA ) -induced hepatotoxicity in rats . Group 1 ( n = 8 ) was the control group . In group 2 ( n = 8 ) , the TAA group , the rats received 300 mg/kg intraperitoneal ( ip ) TAA daily for 2 days . In group 3 ( n = 8 ) , the TAA + DB00065 ( P27352 ) group , infliximab ( 5 mg/kg ip daily ) was administered 48 hours before the first dose of TAA daily for 2 days and was maintained for 4 consecutive days . In group 4 ( n = 8 ) , the P27352 group , the rats received only ip infliximab ( 5 mg/kg ) daily . Livers were excised for histopathological and biochemical tests ( thiobarbituric-acid-reactive substances [ TBARS ] , and myeloperoxidase [ P05164 ] ) . Serum ammonia , aspartate transaminase ( Q9NRA2 ) , alanine transaminase ( ALT ) , P01375 -α , liver TBARS and P05164 levels , and liver necrosis and inflammation scores in the TAA group were significantly higher than in the control and P27352 groups ( all p < 0.01 ) . All parameters except Q9NRA2 were not significantly different between TAA and TAA + P27352 . In conclusion , our results suggest that oxidative stress plays an important role in TAA-induced hepatotoxicity , and infliximab does not improve oxidative liver damage . Reactivation of latent tuberculosis by P01375 blockade : the role of interferon gamma . P01375 ( P01375 ) plays a pathogenic role in psoriasis and rheumatoid arthritis but is essential for host defenses against mycobacteria and other granulomatous pathogens . The risk of reactivation of latent Mycobacterium tuberculosis infection is significantly greater with the P01375 monoclonal antibody infliximab than with the soluble P01375 -receptor etanercept . We have examined the biologic basis of this difference using whole blood culture . DB00065 and adalimumab reduced the proportion of T buciclate-responsive cells by 70 and 50 % , respectively , and suppressed antigen-induced P01579 production by 70 and 64 % . In contrast , etanercept produced no significant effect . The difference between infliximab and etanercept remained whether one compared equal or peak therapeutic drug concentrations , suggesting a relationship to mechanism of action rather than pharmacokinetics . DB00051 and etanercept caused divergent , concentration dependent effects on control of intracellular growth of M. tuberculosis . None of the drugs induced significant levels of apoptosis or necrosis in monocytes or T cells , excluding T-cell death as a mechanism for suppression of antigen-induced responses . P22301 production was equally suppressed by all three drugs , excluding excess P22301 as a regulatory mechanism . The tuberculosis risk posed by infliximab may reflect its combined effects on P01375 and IFNgamma . DB00065 in severe steroid-refractory ulcerative colitis : a pilot study . P01375 -alpha ( P01375 alpha ) -neutralization by infliximab has previously proven efficacious in chronic active Crohn 's disease ( CD ) . We performed an open-label pilot study of a single infusion of 5 mg/kg infliximab in six patients with severe active , steroid-refractory ulcerative colitis ( UC ) . Clinical activity was evaluated according to Lichtiger on days -1 , day 7 , and day 28 . Colonoscopy with biopsy was performed on day -1 and day 7 . All patients showed marked clinical improvement by day 7 ( Lichtiger score 16.3 +/- 0.4 [ day -1 ] vs 4.8 +/- 0.7 [ day 7 ] , p < 0.0001 ) . Four of six patients had long-term remission ( Lichtiger score 7.7 +/- 2.2 [ day 28 ] , P < 0.01 compared to day -1 ) , with a median follow-up of 5.5 months . Colonoscopy confirmed significant healing of endoscopic lesions . The inflammatory infiltrate disappeared on H & E stains , with a marked reduction in infiltrating neutrophils . Semiquantitative evaluation of T and B lymphocytes and macrophages by immunohistochemistry did not reveal major differences compared to pre-treatment . Apoptotic cells in the mucosa were reduced on day 7 . Our data point toward a novel efficacious treatment option in severe steroid-refractory UC and raise the need for controlled trials . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . DB00877 antagonizes P01375 induction of P19320 on endothelial cells by inhibiting mTORC2 . Recruitment of circulating leukocytes into inflamed tissues depends on adhesion molecules expressed by endothelial cells ( ECs ) . Here we report that rapamycin pretreatment reduced the ability of P01375 -treated ECs to capture T cells under conditions of venular flow . This functional change was caused by inhibition of P01375 -induced expression of vascular cell adhesion molecule-1 ( P19320 ) and could be mimicked by knockdown of mammalian target of rapamycin ( P42345 ) or rictor , but not raptor , implicating mTORC2 as the target of rapamycin for this effect . Mechanistically , mTORC2 acts through Akt to repress Raf1- Q02750 /2- P27361 /2 signaling , and inhibition of mTORC2 consequently results in hyperactivation of P27361 /2 . Increased P27361 /2 activity antagonizes P19320 expression by repressing P01375 induction of the transcription factor P10914 . Preventing activation of P27361 /2 reduced the ability of rapamycin to inhibit P01375 -induced P19320 expression . In vivo , rapamycin inhibited mTORC2 activity and potentiated activation of P27361 /2 . These changes correlated with reduced endothelial expression of P01375 -induced P19320 , which was restored via pharmacological inhibition of P27361 /2 . Functionally , rapamycin reduced infiltration of leukocytes into renal glomeruli , an effect which was partially reversed by inhibition of P27361 /2 . These data demonstrate a novel mechanism by which rapamycin modulates the ability of vascular endothelium to mediate inflammation and identifies endothelial mTORC2 as a potential therapeutic target . The renal injury and inflammation caused by ischemia-reperfusion are reduced by genetic inhibition of P01375 -α Q96GN5 : a comparison with infliximab treatment . The role of the tumor necrosis factor ( P01375 ) -α in the pathophysiology of renal ischemia/reperfusion ( I/R ) injury is unclear . We investigate the effects of P01375 -α Q96GN5 gene deletion and infliximab administration on the degree of renal injury induced by I/R . P01375 -α Q96GN5 knockout ( P01375 -αR1KO ) and wild-type ( P01375 -αWT ) mice were subjected to bilateral renal artery occlusion ( 30min ) and reperfusion ( 24h ) . DB00065 ( 10mg/kg subcutaneously , s.c. ) was administered 1h before ischemia . At the end of experiments , urea , creatinine , γGT , and Q9NRA2 were measured to assess renal function and reperfusion injury . Markers of oxidative stress , pro-inflammatory mediators , P35228 , P35354 , and NF-κB signaling pathway were measured . Kidney myeloperoxidase ( P05164 ) activity and malondialdehyde ( MDA ) levels were measured to study polymorphonuclear cell infiltration and lipid peroxidation . P01375 -α Q96GN5 gene deletion and infliximab administration prevented the increase of urea , creatinine , γGT , kidney Q9NRA2 levels , P35228 and P35354 expression , NF-κB translocation , P05164 activity and MDA levels . P01375 -α Q96GN5 gene deletion and infliximab administration lowered the histological evidence of renal damage associated with I/R and caused a reduction of nitrotyrosine suggesting reduced nitrosative stress . Our results demonstrate that P01375 -α plays an important role in I/R injury and put forward the hypothesis that modulation of P01375 -α expression may represent a novel and possible strategy . [ DB00065 ( anti- P01375 ) treatment in patients with adult Still 's disease. Experience in 2 cases ] . Adult Still 's disease is a systemic inflammatory disorder of unknown etiology . First-line treatment for Still 's disease includes nonsteroidal anti-inflammatory drugs and corticosteroids . In refractory cases o when the dose of corticosteroid is unacceptably high , other disease modifying antirheumatic drugs have been used . But recent study showed the efficacy anti- P01375 therapy in adult Sill 's disease refractory to conventional therapy . We report a favourable response to infliximab in two patients who has proved resistant to conventional therapy . DB00065 and insulin resistance . P01308 resistance is the most important pathophysiologic feature of obesity , type 2 diabetes mellitus and prediabetic states . P01375 , a proinflammatory cytokine , plays a pivotal role in the pathogenesis of inflammation-associated insulin resistance during the course of rheumatic diseases . Therapies aimed at neutralizing P01375 , such as the monoclonal antibody infliximab , represent a novel approach for the treatment of rheumatic diseases and allow to obtain significant results in terms of control of the inflammatory process . In this article we reviewed the scientific evidence published in the literature about a potential role of P01375 blockade in improving insulin resistance in non-diabetic rheumatic patients . DB00065 for the treatment of ulcerative colitis . DB00065 ( IFX ) , an anti- P01375 biologic agent , has been demonstrated to offer benefits for the treatment of autoimmune disorders , such as rheumatoid arthritis and Crohn 's disease . Several trials have also investigated the efficacy of IFX for the treatment of ulcerative colitis ( UC ) . IFX was found to be well tolerated . In most trials , IFX treatment was more effective than placebo for patients with moderate , moderate-to-severe or severe UC . However , its place in the treatment algorithms for UC remains to be defined and , to this end , clinical trials comparing IFX treatment to conventional therapies are needed . Donor pre-treatment with everolimus or cyclosporine does not reduce ischaemia-reperfusion injury in a rat kidney transplant model . BACKGROUND : Immunosuppressive agents have been investigated in renal ischaemia-reperfusion injury ( IRI ) and have frequently demonstrated a beneficial effect . Most studies focused on treatment of the recipient at the time of transplantation . Pre-treatment of these organs before injury ( pharmacological pre-conditioning ) may particularly protect these organs . This study aimed to investigate the possible protective effects of donor pre-treatment with cyclosporine ( DB00091 ) or the P42345 inhibitor everolimus or their combination against IRI during renal transplantation in a rat model . METHODS : Donors received vehicle , DB00091 ( 5 mg/kg ) , everolimus ( 0.5 mg/kg ) or CsA + everolimus . Two oral doses were administered to the donors at 24 h and again at 6 h prior to donor kidney removal . Syngeneic rat kidneys were preserved in UW solution for 24 h prior to transplantation . After 24 h of reperfusion , blood and tissue samples were collected from recipients for further analysis . RESULTS : Renal functions as determined by creatinine and necrosis scores were not different between the experimental groups . Cleaved caspase-3 , heat shock protein 70 ( HSP70 ) , tumor-necrosis factor-alpha ( P01375 -α ) and nitrotyrosine protein levels were not statistically different between the four treatment groups at 24 h post-transplantation . Blood NMR analysis on metabolic markers for IRI reveals no beneficial effects of donor pre-treatment on the 24-h outcome in transplantation . CONCLUSIONS : When given alone or as a combination to donors before organ recovery , cyclosporine or everolimus does not appear to ameliorate IRI . Aerosol vaccination with AERAS-402 elicits robust cellular immune responses in the lungs of rhesus macaques but fails to protect against high-dose Mycobacterium tuberculosis challenge . Development of a vaccine against pulmonary tuberculosis may require immunization strategies that induce a high frequency of Ag-specific P01730 and CD8 T cells in the lung . The nonhuman primate model is essential for testing such approaches because it has predictive value for how vaccines elicit responses in humans . In this study , we used an aerosol vaccination strategy to administer AERAS-402 , a replication-defective recombinant adenovirus ( rAd ) type 35 expressing Mycobacterium tuberculosis Ags Ag85A , Ag85B , and TB10.4 , in bacillus Calmette-Guérin ( BCG ) -primed or unprimed rhesus macaques . Immunization with BCG generated low purified protein derivative-specific P01730 T cell responses in blood and bronchoalveolar lavage . In contrast , aerosolized AERAS-402 alone or following BCG induced potent and stable Ag85A/b-specific P01730 and CD8 effector T cells in bronchoalveolar lavage that largely produced IFN-γ , as well as P01375 and P60568 . Such responses induced by BCG , AERAS-402 , or both failed to confer overall protection following challenge with 275 CFUs M. tuberculosis Erdman , although vaccine-induced responses associated with reduced pathology were observed in some animals . Anamnestic T cell responses to Ag85A/b were not detected in blood of immunized animals after challenge . Overall , our data suggest that a high M. tuberculosis challenge dose may be a critical factor in limiting vaccine efficacy in this model . However , the ability of aerosol rAd immunization to generate potent cellular immunity in the lung suggests that using different or more immunogens , alternative rAd serotypes with enhanced immunogenicity , and a physiological challenge dose may achieve protection against M. tuberculosis . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . DB00065 for steroid-refractory acute GVHD : a case series . Acute and chronic graft-versus-host disease ( GVHD ) remain major barriers to successful hematopoietic stem cell transplantation ( P09683 ) . P01375 has been implicated in the pathogenesis of GVHD and P01375 blockade has been explored for treatment of GVHD . The development of a chimeric mouse/human monoclonal antibody ( infliximab ) which binds to cells producing P01375 , allowing for not only the neutralization of P01375 but also lysis of the cells producing the P01375 , makes this an attractive drug to explore in GVHD . We report on 11 patients with acute GVHD who were treated with infliximab after failing other therapies . The survival was very poor , in keeping with previously published reports of steroid-refractory acute GVHD . Two patients with severe diarrhea from acute GI GVHD resolved their symptoms after treatment with infliximab . Only these two patients survived . It appears that of all acute GVHD manifestations , gastrointestinal GVHD may be more responsive to treatment with infliximab than others . Caution is recommended when using this agent since it may exacerbate active infections , particularly aspergillosis . Furthermore , we do not know the correct dose or schedule to use with this drug . Given these data , controlled studies assessing dose and timing of administration may be warranted to study infliximab in acute GVHD . Integrating anti-tumor necrosis factor therapy in inflammatory bowel disease : current and future perspectives . Crohn 's disease and ulcerative colitis are two idiopathic inflammatory disorders of the GI tract . Manifestations of disease can be severe and lead to long term therapy with a variety of medications and/or surgery . Standard medical therapy consists of agents that either treat suppurative complications or modulate the inflammatory cascade in a nonspecific manner . Many specific chemokine and cytokine effectors that promote intestinal inflammation have been identified . Such work has led to experimental clinical trials with a variety of cytokine antagonists . Compounds directed against one such cytokine , tumor necrosis factor alpha ( P01375 ) , have demonstrated the greatest clinical efficacy to date . This is consistent with scientific observations that suggest a central role for P01375 in the inflammatory cascade . DB00065 is a chimeric monoclonal antibody against P01375 that has been demonstrated to be effective for the treatment of Crohn 's disease . DB00065 is Food and Drug Administration approved for the treatment of Crohn 's disease . There exist several other P01375 antagonists in various phases of investigation , including the monoclonal antibody CDP 571 , the fusion peptide etanercept , the phosphodiesterase inhibitor oxpentifylline , and thalidomide . The clinical efficacy of these agents and the role of P01375 in the pathogenesis of inflammatory bowel disease is reviewed . [ The role of anti- P01375 therapy in ulcerative colitis ] . Anti- P01375 -alfa molecules are currently being used to treat ulcerative colitis regarding to the fact that P01375 has an important role in the pathogenesis of Q9UKU7 . Although these drugs improved the therapy of patients , immunogenicity limits their potential for clinical use . DB00065 and adalimumab are effective for induction and maintenance of remission in outpatients with moderate to severe steroid-refractory ulcerative colitis . Biologics can be a drug of choice for patients with refractory proctitis and refractory pouchitis . In hospitalized patients with steroid-resistant severe ulcerative colitis who are candidates for colectomy , infliximab may be second-line option . Adequate long-term maintenance therapy with anti- P01375 is required after rescue therapy for a sustained benefit . Regarding to the known risk for side-effects of anti- P01375 drugs especially in patients concomitantly treated with thiopurines it is urgent future research . DB00065 for the treatment of orofacial Crohn 's disease . Orofacial manifestations of Crohn 's disease can be difficult to diagnose and treat . We report a case in which the orofacial lesions occurred 7 years prior to the diagnosis of underlying inflammatory bowel disease . The patient was refractory to mesalamine and systemic corticosteroids but responded to infliximab , the chimeric monoclonal antibody to tumor necrosis factor ( P01375 ) . A review of the literature of the orofacial granulomatoses is presented as well . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Tuberculosis following the use of etanercept , a tumor necrosis factor inhibitor . DB00065 , a tumor necrosis factor ( P01375 ) antagonist , is associated with tuberculosis ( TB ) , but it is unknown whether this phenomenon is true of all P01375 antagonists . We reviewed 25 cases of TB due to another P01375 antagonist , etanercept , that were reported to the US Food and Drug Administration ( FDA ) between November 1998 and March 2002 . Such cases are sometimes incomplete and are subject to underreporting . Fifteen patients received other immunosuppressive medications . The median interval between the receipt of the first dose of etanercept and the diagnosis of TB was 11.5 months . Thirteen patients had extrapulmonary TB at the time of diagnosis . Diagnosis was made on the basis of culture results for 12 patients , biopsy findings for 9 , and sputum staining for 4 . There were 2 deaths , 1 of which was directly attributed to TB . The estimated number of TB cases reported to the FDA for each person-year of treatment with etanercept ( i.e. , the " reporting rate " ) among patients with rheumatoid arthritis ( RA ) was ~10 cases/100,000 patient-years of exposure . Clinicians considering etanercept for patients with RA should be alert to the possibility of the occurrence of TB , sometimes with an unusual extrapulmonary presentation . It is unclear whether etanercept therapy increases the risk of TB beyond the elevated TB rates already documented for patients with RA . Circulating apoptotic proteins are increased in long-term disease-free breast cancer survivors . Circulating apoptotic proteins are increased in patients with heart failure . We evaluated whether circulating soluble ( s ) apoptosis-related proteins and inflammation markers are increased in long-term disease free breast cancer survivors and associated with cardiotoxicity , and if subgroups could be identified based on the applied treatments . Circulating tumour necrosis factor ( P01375 ) alpha , sTNF-receptor ( sTNF-R ) 1 and 2 , sFas , sFas ligand , sTNF-related apoptosis inducing ligand ( sTRAIL ) and serum P04626 were measured with immunoassay . High-sensitivity P02741 ( HS-CRP ) , fibrinogen , plasma B-type and N-terminal atrial natriuretic peptide ( NT- P01160 and DB04899 ) were also determined . Thirty-four patients with median 6.0 years follow-up and 12 healthy age-matched women were enrolled . Chemotherapy , consisting of five cycles fluorouracil , epirubicin ( 90 mg/m(2) ) , cyclophosphamide ( FEC ) ( n=14 ) or four cycles FEC followed by myeloablation with high-dose carboplatin , cyclophosphamide , thiotepa ( n=20 ) , preceded irradiation and tamoxifen . Circulating apoptosis markers were higher in patients than in controls . No associations with cardiac dysfunction were observed . sFas ligand and sTRAIL were higher in the high-dose than in the standard-dose group . In conclusion , we observed increased circulating apoptotic protein levels in long-term disease-free breast cancer survivors , treated with adjuvant chemoradiotherapy , particularly after myeloablative chemotherapy . The potential relation with late cardiotoxicity of antineoplastic therapy deserves further study . Phenotype and functional changes of Vgamma9/Vdelta2 T lymphocytes in Behçet 's disease and the effect of infliximab on Vgamma9/Vdelta2 T cell expansion , activation and cytotoxicity . INTRODUCTION : DB00065 is a chimeric monoclonal antibody against tumor necrosis factor alpha ( P01375 ) that has been introduced recently for Behçet 's disease ( BD ) patients who were resistant to standard treatment . The aim of this study was to analyse the functional changes of Vgamma9/Vdelta2 T lymphocytes in both active and inactive disease and the effect of infliximab on Vgamma9/Vdelta2 T cell expansion , activation and cytotoxicity . METHODS : We investigated 1 ) cell expansion , 2 ) expression of TNFRII receptor , 3 ) perforin and gamma interferon ( IFN ) content , 4 ) release of granzyme A ( GrA ) and 5 ) phenotype changes , in vitro and in vivo , in Vgamma9/Vdelta2 T lymphocytes by means of fluorescence-activated cell sorter analysis of lymphocyte cultures from patients with active and inactive BD and healthy subjects . RESULTS : Cell expansion , expression of TNFRII , perforin and gamma IFN content and release of granzyme A were significantly higher in active patients . In vitro and ex vivo treatment with infliximab resulted in a significant reduction of all parameters together with changes in the phenotype of Vgamma9/Vdelta2 T cells . CONCLUSIONS : All together these data indicate that infliximab is capable of interfering with Vgamma9/Vdelta2 T cell function in BD and although cell culture models can not reliably predict all potential effects of the drug in vivo , our results present the possibility that this drug may find use in a range of immunological disorders , characterized by dysregulated cell-mediated immunity . [ A case of psoriasis induced by infliximab treatment for Crohn 's disease ] . DB00065 , the monoclonal antibody to tumor necrosis factor , is indicated for refractory luminal and fistulizing Crohn 's disease and rheumatoid arthritis . DB00065 treatment has adverse events including infusion reactions , opportunistic infections , and the potential for the event such as reactivation of latent tuberculosis . Cutaneous adverse reactions of P01375 -α agents include skin rash , urticaria , pruritus , lupus-like eruption , and injection site reactions . Most of all , psoriasis or psoriasiform dermatitis induced by infliximab treatment for Crohn 's disease is rarely reported in Korea . We report a case of psoriasis induced by infliximab treatment for Crohn 's disease with a review of world literature . DB00065 induced T lymphocyte apoptosis in Crohn 's disease . Crohn 's disease ( CD ) is a chronic inflammatory disorder of the gastrointestinal tract of unknown origin . Therapies include immune modulating agents , biological therapies , and surgery . The activity and efficacy of the anti-tumor necrosis factor ( P01375 ) therapies infliximab and etanercept have proved to be different : infliximab is effective to induce and maintain remission in refractory CD , while etanercept is not . This brief review considers the question of whether this disparity can be explained by the different structure of the proteins , their different binding affinities , or the subsequent effects on T lymphocytes . [ Intestines and joints : an entity ? ] . The intestinal inflammation and the inflammatory arthropathy in Crohn 's disease and in spondyloarthropathy are tightly linked . Both forms of inflammation occur in clinical and subclinical forms in both diseases : 60 % of patients with Crohn 's disease have clinical and/or radiological signs of articular involvement -- 65 % of patients belonging to the concept of spondyloarthropathy have endoscopic and/or histological signs of gut inflammation . The macrophages and the lymphocytes play a role either by the transport of bacterial antigens by macrophages from the gut to the synovium , either by the recirculation of activated lymphocytes attracted to the tissue by adhesion molecules . In both diseases , P01375 alpha play a central role resulting in a rapid and spectacular improvement of the intestinal and articular inflammation by the administration of a monoclonal chimeric antibody directed against this cytokine ( DB00065 ) . This clinical effect is associated or secondary to a restoration of the production of proinflammatory cytokines . According the importance of genetic factors in the etiopathogenesis of both diseases ( mutation in the Q9HC29 gene for Crohn -- HLA- Q8TCY5 phenotype for spondyloarthropathy ) future studies are necessary to find a common alteration explaining the link between gut and joint . This knowledge can help in the elucidation of the origin of both diseases . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . DB00065 treatment in a case of rheumatoid scleromalacia perforans . Ocular rheumatoid disease manifests as hyperemia of the conjunctiva and episclera , and in severe cases , episcleritis can result in nodular sclerotic and scleromalacia perforans . A clinical case of scleromalacia perforans in a 56-year-old woman with 20 years of seropositive rheumatoid arthritis of functional class IV is presented here . During that period , she received exclusively non-steroidal anti-inflammatory drugs ( NSAIDs ) . She developed acute episcleritis of the left ocular globe , which rapidly progressed to scleromalacia perforans . Since the left eye became perforated , it was surgically enucleated , and the patient was maintained with steroidal therapy . Nevertheless , two months later she developed new-onset episcleritis of the right eye followed by scleromalacia . She was first evaluated by a rheumatologist and treated with 200 mg/dose of infliximab , which was administered monthly for the following four months . The biological treatment was accompanied by methotrexate and prednisone . With this therapy , the ocular lesion dramatically improved , and complete remission of rheumatoid arthritis and scleritis was archived four months later . In conclusion , tumour necrosis factor ( P01375 ) blockers are effective therapeutic agents in ocular complications of rheumatoid arthritis . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Short-term P01375 inhibition reduces short-term and long-term inflammatory changes post-ischemia/reperfusion in rat intestinal transplantation . BACKGROUND : P01375 ( P01375 ) -α inhibition was shown to reduce ischemia/reperfusion injury ( IRI ) after intestinal transplantation ( ITX ) . We studied the effects of different TNFα inhibitors on acute IRI and long-term inflammatory responses in experimental ITX . METHODS : Orthotopic ITX was performed in an isogenic ischemia/reperfusion model in Lewis rats . The TNFα inhibition groups received infliximab post-reperfusion ; etanercept pre-reperfusion and at postoperative days ( POD ) 1 , 3 , 5 , and 7 ; or pentoxifylline pre-reperfusion and at POD 1 to 5 . Tissue samples were taken from proximal and distal graft sections and mesenteric lymph nodes at 20 min , 12 hr , 7 day , and 6 months post-reperfusion for histopathology , immunohistology , terminal deoxyribosyl transferase-mediated dUTP nick-end labeling ( TUNEL ) assay , and real-time RT-PCR . Lung sections were stained for the myeloperoxidase assay . RESULTS : TNFα inhibitors decreased inflammatory changes after IRI in all treatment groups . DB00065 significantly improved 7-day survival and reduced the histological and immunohistochemical signs of IRI , the numbers of graft-infiltrating T cells and Q92838 monocytes and macrophages , and pulmonary neutrophil infiltration , and also enhanced the accumulation of cytoprotective markers . Graft injury was more prominent in the distal graft than in the proximal graft in all groups , regardless of TNFα inhibition . CONCLUSION : DB00065 significantly reduced both acute IRI and , as with other TNFα inhibitors , long-term inflammatory responses after rat ITX . TNFα inhibition may help diminish chronic inflammatory long-term effects and avoid chronic allograft enteropathy . Redo Ileal pouch-anal anastomosis combined with anti- P01375 -α maintenance therapy for Crohn 's disease with pelvic fistula : report of two cases . Pouch failure has been reported to occur after ileal pouch-anal anastomosis for Crohn 's disease . We report two cases of patients with Crohn 's disease , who underwent redo ileal pouch-anal anastomosis ( redo-IPAA ) combined with anti- P01375 -α maintenance therapy , with good functional results . The first patient , a man with presumed ulcerative colitis , suffered pelvic fistula recurrence and anastomotic dehiscence . He underwent redo-IPAA , at which time longitudinal ulcers were found . DB00065 was started 4 days postoperatively and continued . The second patient , a woman treated for ulcerative colitis , underwent laparoscopic IPAA 8 years later . After the development of a pelvic fistula , twisted mesentery of the ileal pouch was found intraoperatively and Crohn 's disease was diagnosed . DB00051 therapy resulted in fistula closure . Redo-IPAA was performed to normalize the twisted mesentery of the ileal pouch . No complications have been observed in either patient , both of whom have experienced good functional results after closure of the covering stomas . T cell lymphoproliferative disorders associated with anti-tumor necrosis factor alpha antibody therapy for ulcerative colitis : literature summary . The enhanced risk of development of lymphoproliferative disorders in patients with inflammatory bowel disease has been attributed to immunosuppressive/immunomodulatory therapies . DB00065 is a chimeric monoclonal immunoglobulin P55008 antibody directed against tumor necrosis factor alpha ( P01375 -α ) that was approved by the Food and Drug Administration ( FDA ) in 1998 as an effective therapeutic agent against inflammatory bowel disease . Malignant lymphomas of both B and T cell lineage have been described in patients undergoing therapy involving P01375 -α blockade . To date , eight cases of Epstein-Barr virus ( EBV ) -negative hepatosplenic T cell lymphoma associated with infliximab have been reported to the FDA 's Adverse Event Reporting System , as well as several other T cell lymphoproliferative disorders with aggressive clinical outcomes . We present the histologic , immunophenotypic , and molecular features of a T cell lymphoproliferative disorder involving the axillary lymph node of a 33-year-old male following infliximab treatment for ulcerative colitis . These EBV-negative lymphomas suggest that lymphoproliferative disorders following infliximab treatment for inflammatory bowel disease may involve EBV-independent immune dysregulation . The spectrum of lymphoproliferative disorders associated with infliximab and the potential mechanisms by which they occur are discussed . [ The importance of biologicals in the treatment of SoJIA ] . Systemic onset juvenile idiopathic arthritis ( SoJIA ) remains difficult to treat . In addition to conventional antirheumatic therapy with non-steroidal antirheumatic drugs ( NSARDs ) , steroids or disease-modifying antirheumatic drugs ( DMARDs ) , biologicals offer a new therapeutic approach for this disease in that they are able to target pathogenically relevant cytokines and effector cells . Some biologicals are already approved for use in children with rheumatic disease.In order to assess the currently available data on the use of biologicals in SoJIA , we performed a Medline search for the period 2005 to March 2010 , including the MeSH terms " SoJIA " , " systemic juvenile idiopathic arthritis " and " biologicals " , as well as an NIH study registry search . At Present there are scant and unconvincing data on the use of DB00005 or DB00051 for the treatment of SoJIA . No results are published on the use of DB00065 or other new P01375 inhibitors . The inhibition of IL-1 or P05231 shows promising results . Data on the efficacy of DB01281 is limited due to very low numbers of SoJIA patients in the studies.Further studies on the use of biologicals in SoJIA while taking individual factors into consideration are required . The long-term safety of all biologicals should be investigated in prospective registers . Oral ulcerations are associated with the loss of response to infliximab in Crohn 's disease . We describe a 25-year-old Caucasian man with a 13-year history of inflammatory Crohn 's disease ( CD ) who was suffering recurrent severe oral and esophageal ulcerations for the past 3 years . DB00117 CD had been treated with infliximab infusions among other medications . The loss of efficacy was confirmed by antibodies to infliximab ( ATI ) and serum infliximab tests that showed high levels of ATIs and undetectable levels of infliximab respectively . These findings were consistent with significant immunogenic response to infliximab leading to loss of effect . DB00065 infusions and prednisone were discontinued and treatment of the CD was instituted with adalimumab , a human anti-tumor necrosis factor ( P01375 ) -alpha biologic agent , to control the inflammatory small intestinal disease and dapsone for the oral and esophageal CD ulcerations . The patient 's oral and esophageal lesions as well as the enteric CD are under control after 5 months of therapy . DB00065 modifies mesenteric adipose tissue alterations and intestinal inflammation in rats with TNBS-induced colitis . OBJECTIVE : DB00065 is a monoclonal anti- P01375 -α antibody that is used therapeutically to treat Crohn 's disease ( CD ) . High levels of pro-inflammatory cytokines , especially P01375 -α , have been observed in the gastrointestinal tract of CD patients and were associated with alterations in the mesenteric adipose tissue , which also contributed to the high levels of adipokine release . The authors used a rat model of colitis that produces mesenteric adipose tissue alterations that are associated with intestinal inflammation to study the effects that infliximab treatment has on adipokine production , morphological alterations in adipose tissue and intestinal inflammation . MATERIAL AND METHODS : The ability of infliximab to neutralize rat P01375 -α was evaluated in vitro using U937 cells . Colitis was induced by repeated intracolonic trinitrobenzene sulfonic acid instillations and was evaluated by macroscopic score , histopathological analysis , myeloperoxidase activity , P01375 -α and P22301 expression as well as P35228 ( inducible NO synthase ) expression and JNK phosphorylation in colon samples . The alterations in adipose tissue were assessed by P01375 -α , P22301 , leptin , adiponectin and resistin levels as well as adipocyte size and peroxisome proliferator-activated receptor ( Q07869 ) -γ expression . RESULTS : DB00065 treatment controlled intestinal inflammation , which reduced lesions and neutrophil infiltration . Inflammatory markers , such as P35228 expression and JNK phosphorylation , were also reduced . In mesenteric adipose tissue , infliximab increased the production of P22301 and resistin , which was associated with the restoration of adipocyte morphology and Q07869 -γ expression . CONCLUSIONS : Our results suggest that infliximab could contribute to the control of intestinal inflammation by modifying adipokine production by mesenteric adipose tissue . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . [ DB00065 ( chimeric monoclonal antibody to tumor necrosis factor alpha ) ] . In rheumatoid arthritis and inflammatory bowel disease , such as Crohn 's disease , certain immunological abnormalities are considered as its cause , but the fundamental mechanism remains unclear . However , recent researches revealed that inflammatory cytokines , such as P01375 , IL-1 and P05231 , are greatly involved in these immunological abnormalities . This led to the development of anti-cytokine therapy using monoclonal antibodies to these cytokines . Among them , DB00065 , a chimeric monoclonal antibodies to P01375 , is most clinically applied and marked therapeutic effect is seen in various diseases . [ Proteomic analysis of changes in the serum protein profile by anti- P01375 therapy ] . We analyzed the alternation in serum protein by infliximab therapy using proteomics-based technique . More than 50 gel spots were seen to increase or decrease in correlation with clinical improvements of rheumatoid arthritis ( RA ) . Spots of interest were identified by two dimensional electrophoresis and mass spectrometry . DB00065 therapy reduced the inflammatory proteins such as P02741 , serum amyloid protein A , serum amyloid protein P , and alpha1-acid glycoprotein , while the therapy increased Apo A-I , retinol-binding protein , vitamin D-binding protein , and gelsolin . This suggested that infliximab therapy shifted the inflammatory status of serum protein profile of RA patients to normal and modified the extracellular actin-scavenging system as well as vitamin and lipid profile . P01375 alpha blockade reduces the synovial cell infiltrate early after initiation of treatment , but apparently not by induction of apoptosis in synovial tissue . OBJECTIVE : To determine whether treatment with the chimeric anti-tumor necrosis factor alpha antibody infliximab could reduce cellularity by the induction of apoptosis in synovial tissue . METHODS : Twenty-four rheumatoid arthritis patients with active disease were randomized to receive either infliximab ( 3 mg/kg ) ( n = 12 ) or placebo ( n = 12 ) intravenously . All patients were subjected to arthroscopic synovial biopsy directly before initiation of treatment . A second arthroscopic synovial biopsy of the same index joint was performed 48 hours after the first arthroscopy . After the second arthroscopy , the patients who had initially received placebo were also treated with infliximab in an extension study . A third arthroscopy was performed in all patients on day 28 . Immunohistologic analysis was performed to characterize the cell infiltrate . In situ detection of apoptotic cells was performed by TUNEL assay and electron microscopy . RESULTS : At 48 hours after initiation of infliximab treatment , there was a significant reduction in the number of intimal macrophages ; this was not observed in the placebo group . The number of sublining macrophages , T cells , and plasma cells also tended to be decreased in infliximab-treated patients , but not in the placebo group . Of interest , we did not detect any increase in the number of apoptotic cells after infliximab treatment . CONCLUSION : DB00065 therapy may reduce the number of inflammatory cells in rheumatoid synovial tissue as soon as 48 hours after initiation of treatment , but apparently not by induction of apoptosis . Conceivably , decreased cell infiltration primarily results from early inhibition of cell migration . Treatment of pyoderma gangrenosum with infliximab in Crohn 's disease . Pyoderma gangrenosum ( PG ) is an ulcerating noninfectious disease of the skin seen in 1 to 5 % of patients with inflammatory bowel disease . The pathogenesis of PG has yet to be determined but may be related to abnormal T cell responses and the production of P01375 , a powerful proinflammatory cytokine . DB00065 , a chimeric monoclonal antibody to P01375 , has been approved for the treatment of Crohn 's disease . We present four patients with PG treated with DB00065 for fistulizing Crohn 's in whom complete healing of PG was achieved . Four patients with active fistulizing Crohn 's disease and PG were treated . All patients were females ranging in age from 48 to 60 years , with a mean age of 54 years . Three of four patients had PG lesions located on the lower extremities ; one patient had peristomal disease . All patients had at least colonic involvement of their Crohn 's . The patients received either a single infusion or a series of three 5 mg/kg DB00065 infusions . All four patients demonstrated rapid healing of PG within 4 weeks of the first infusion of DB00065 . PG healing followed improvement in bowel disease . Complete resolution without recurrence was noted in all patients . Rapid resolution of PG was noted in four female patients with fistulizing Crohn 's disease treated with DB00065 . Healing was complete , without recurrence . The anti- P01375 properties of DB00065 suggest that healing may be mediated by the drug 's effect on cytokine pathways , perhaps by blunted T cell activation early in the inflammatory cascade . We suggest an independent effect of DB00065 on PG . Interferon-induced increase in sensitivity of ovarian cancer targets to lysis by lymphokine-activated killer cells : selective effects on P04626 /neu-overexpressing cells . Overexpression of the P04626 /neu oncogene in ovarian tumor cells is associated with relative resistance to lymphokine-activated killer ( Q96QP1 ) cell cytotoxicity . Treatment with gamma-interferon ( P01579 ) ( 200-2000 units/ml ) for 3 days markedly enhanced the sensitivity of P04626 /neu-overexpressing ovarian tumor cells to Q96QP1 cells but had no effect on the sensitivity of nonexpressing ovarian targets . Increased sensitivity to lysis was associated with an increase in effector-target conjugate formation , the induction of target cell intercellular adhesion molecule 1 ( P05362 ) expression , and the down-regulation of P04626 /neu expression . Anti- P05362 antibody blocked the enhanced lysis , indicating that P05362 is important in the increased sensitivity to Q96QP1 cells . However , induction of P05362 expression did not correlate well with enhanced sensitivity to lysis ; it was maximal after 24 h of exposure to P01579 and still present 24 h after removing P01579 . In contrast , enhanced lysis required 3 days of exposure to P01579 and was reversed within 24 h after removal of P01579 . These data indicate that , although P05362 is necessary , it is not sufficient for the P01579 -induced enhancement of sensitivity to Q96QP1 lysis . Disseminated tuberculosis infection and paradoxical reaction during antimycobacterial treatment related to P01375 blocker agent DB00065 . P01375 ( P01375 ) -alpha inhibitors play an important role in the treatment of immun-mediated diseases such as Crohn 's disease . But they also have been related to increased risk for disseminated Mycobacterium tuberculosis infections and paradoxical response to antimycobacterial treatment . Here we report a disseminated tuberculosis case and a paradoxical response to treatment after receiving P01375 -inhibitor agent DB00065 for Crohn 's disease . The patient had a severe clinical condition before the antimycobacterial treatment and although proper treatment was initiated his radiological findings were worsened one month after initiation of the treatment . All control microbologic tests for secondary infections were negative and this situation was accepted as a paradoxical response to antimycobacterial treatment and treatment was continued with the same regimen . At the end of the second month of the treatment , most of the symptoms disappeared and chest radiograph findings were better than the previous one . In conclusion , P01375 inhibitor therapy increases risk of mycobacterial infections and patients should be examined carefully about tuberculosis before starting this therapy . Also , it is important for physicians to recognize and know how to manage paradoxical response related to P01375 inhibitors during anti-tuberculosis treatment . DB00065 in patients who have spondyloarthropathy : clinical efficacy , safety , and biological immunomodulation . A major breakthrough has been achieved in the treatment of patients who have AS and other types of SpA . The identification of the expression and role of P01375 in patients who have these diseases and the recognition of their relation with gut inflammation ( where infliximab therapy has proven efficacious already ) has led to the successful use of P01375 blockade in SpA , establishing a new indication for this type of anticytokine therapy . Evidence supports equal response in cases of axial or peripheral disease . DB00065 therapy has been most extensively documented in this new indication for anti- P01375 therapy , but other compounds are also in the field . Gorman et al reported on 40 patients who had active AS who were randomly assigned to receive twice-weekly subcutaneous injections of etanercept ( 25 mg ) or placebo for 4 months [ 65 ] . The primary endpoint was a composite of improvements . Treatment with etanercept resulted in significant and sustained improvement ( treatment response in 80 % in the etanercept group versus 30 % in the placebo ) . Data regarding the human anti- P01375 monoclonal antibody adalimumab in SpA are not yet available . Different questions remain open , including optimal dosing , long-term safety , and effects of this new treatment on the structural articular level ; however , a therapeutic breakthrough like the one currently reviewed has seldom occurred in arthritis care . Antiinflammatory steroid action in human ovarian surface epithelial cells . The human ovarian surface epithelium ( OSE ) is subject to serial injury and repair during ovulation , which is a natural inflammatory event . We asked whether there is a compensatory antiinflammatory component to this process , involving steroid hormones produced locally at the time of ovulation . Quantitative RT-PCR analysis of total RNA from cultured human OSE cell monolayers showed that exposure to proinflammatory IL1alpha ( 500 pg/ml ) increased mRNA levels of cyclooxygenase-2 ( P35354 ) ( P < 0.01 ) at 48 h . The P35354 mRNA response to IL1alpha was associated with an approximate 18-fold ( P < 0.01 ) increase in mRNA levels of 11beta-hydroxysteroid dehydrogenase type 1 ( 11betaHSD1 ) , encoding the steroid dehydrogenase that reversibly reduces cortisone to antiinflammatory cortisol . Addition of cortisol to OSE cell culture medium dose-dependently suppressed the P35354 mRNA response to IL1alpha ( P < 0.01 ) but reciprocally enhanced the 11betaHSD1 mRNA response ( P < 0.05 ) , with both effects strongest at 1 microm cortisol . Presence of glucocorticoid receptor-alpha mRNA and protein was established in OSE cell monolayers and treatment with IL1alpha shown to significantly up-regulate the glucocorticoid receptor-alpha mRNA level ( P < 0.05 ) . P04150 antagonist ( DB00834 , 10 microm ) fully reversed the inhibitory effect of 1 microm cortisol on IL1alpha-stimulated P35354 mRNA expression . Progesterone also suppressed IL1alpha-induced P35354 mRNA expression but had no significant effect on IL1alpha-stimulated 11betaHSD1 expression . These data provide direct evidence for antiinflammatory actions of cortisol and progesterone in human OSE cells . Inhibition of JAKs in macrophages increases lipopolysaccharide-induced cytokine production by blocking P22301 -mediated feedback . Macrophages are an important source of cytokines following infection . Stimulation of macrophages with TLR agonists results in the secretion of P01375 -α , P05231 , and IL-12 , and the production of these cytokines is controlled by multiple feedback pathways . Macrophages also produce P22301 , which acts to inhibit proinflammatory cytokine production by macrophages via a JAK/ P40763 -dependent pathway . We show in this paper that , DB08877 , a recently described selective inhibitor of JAKs , increases P01375 , P05231 , and IL-12 secretion in mouse bone marrow-derived macrophages stimulated with LPS . This effect is largely due to its ability to block P22301 -mediated feedback inhibition on cytokine transcription in macrophages . Similar results were also obtained with a second structurally unrelated Jak inhibitor , DB08895 . In addition , LPS induced the production of IFN-β , which was then able to activate JAKs in macrophages , resulting in the stimulation of P42224 phosphorylation . The initial induction of P22301 was independent of JAK signaling ; however , inhibition of JAKs did reduce P22301 secretion at later time points . This reflected a requirement for the IFN-β feedback loop to sustain P22301 transcription following LPS stimulation . In addition to P22301 , IFN-β also helped sustain P05231 and IL-12 transcription . Overall , these results suggest that inhibition of JAKs may increase the inflammatory potential of macrophages stimulated with O00206 agonists . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . Can we modulate the clinical course of inflammatory bowel diseases by our current treatment strategies ? Ulcerative colitis and Crohn 's disease are chronic disabling lifelong diseases which may be disturbed by severe flares and anatomical complications requiring surgery . Until the very last years there was no clear indication that treatment was able to modify the long-term natural history of the disease . In particular , there are no data demonstrating a clear improvement through the period 1950-2003 in disease activity , occurrence of complications and need for surgery , in spite of an increased use of immunosuppressants since the 1990s . However , in inflammatory bowel disease , both thiopurines and methotrexate are very efficient in about one half of the patients , and in responders , may heal the mucosa and decrease the need for surgery . The early use of immunosuppressants in selected patients may have an impact on occurrence of severe flares and complications , and need for surgery . Moreover , anti- P01375 now used for 10 years in Crohn 's disease and for 5 years in ulcerative colitis demonstrated in two thirds of the patients a remarkable anatomic effect , healing the mucosa , closing fistulae and preventing strictures . DB00065 does prevent endoscopic recurrence following ileal resection for Crohn 's disease . Actually , because irreversible anatomical damage may develop during the first years of disease , there is a need to classify early in the course of the disease patients who will benefit from anti- P01375 and classical immunosuppressants , respectively . There is the need in the next few years to better define these subgroups and to compare different strategies within each group through randomized interventional studies . Alopecia areata universalis during off-label treatment with DB00065 in a patient with Behçet disease . DB00065 , a chimeric monoclonal anti- P01375 -alfa agent used to treat autoimmune diseases , has shown a paradoxical side effect in the development of autoimmunity . We describe a case of alopecia areata universalis associated with infliximab treatment in a patient with Behçet disease . This case suggests a complex and contradictory role of P01375 -α in the pathogenesis of alopecia areata . DB00065 for treatment of resistant pyoderma gangrenosum associated with Crohn 's disease . We present the case of an 18-year-old woman with Crohn 's disease manifested by diffuse abdominal pain , bloody diarrhea accompanied by arthralgia , and swelling of large joints . On the lateral aspect of her right ankle there was an hemorrhagic , necrotic bullous lesion measuring 3 x 4 cm , surrounded by cutaneous inflammation and erythema . Biopsy showed a neutrophilic abscess-like ulcerative skin inflammation , which was diagnosed as pyoderma gangrenosum . The patient was treated with high doses of parenteral methylprednisolone , but her condition failed to improve and infliximab , a P01375 blocking agent , was instituted . An immediate response of Crohn 's disease was observed and , over the next 5 weeks , the ulcer on her right ankle also healed completely . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Comparison of drug survival rates for tumor necrosis factor antagonists in rheumatoid arthritis . BACKGROUND : Persistence of anti-tumor necrosis factor ( P01375 ) therapy in rheumatoid arthritis ( RA ) is an overall marker of treatment success . OBJECTIVE : To assess the survival of anti- P01375 treatment and to define the potential predictors of drug discontinuation in RA , in order to verify the adequacy of current practices . DESIGN : An observational , descriptive , longitudinal , retrospective study . SETTING : The Hospital Clínico Universitario de Valladolid , Valladolid , Spain . PATIENTS : RA patients treated with anti- P01375 therapy between January 2011 and January 2012 . MEASUREMENTS : Demographic information and therapy assessments were gathered from medical and pharmaceutical records . Data is expressed as means ( standard deviations ) for quantitative variables and frequency distribution for qualitative variables . Kaplan-Meier survival analysis was used to assess persistence , and Cox multivariate regression models were used to assess potential predictors of treatment discontinuation . RESULTS : In total , 126 treatment series with infliximab ( n = 53 ) , etanercept ( n = 51 ) or adalimumab ( n = 22 ) were administered to 91 patients . DB00065 has mostly been used as a first-line treatment , but it was the drug with the shortest time until a change of treatment . Significant predictors of drug survival were : age ; the anti- P01375 agent ; and the previous response to an anti- P01375 drug . LIMITATION : The small sample size . CONCLUSION : The overall efficacy of anti- P01375 drugs diminishes with time , with infliximab having the shortest time until a change of treatment . The management of biologic therapy in patients with RA should be reconsidered in order to achieve disease control with a reduction in costs . Anti- P01375 treatment in Crohn 's disease : toward tailored therapy ? DB00065 is a potent therapy for induction and maintenance of remission in Crohn 's disease . Unfortunately , many patients lose response and/or develop allergic reactions caused by the chimeric antibody . By means of measuring the presence of antibodies against infliximab and the trough levels of the drug , it seems easier to predict whether patients will respond to intensified dose regimens or will rather benefit from a switch to alternative agents . B cell activation in rheumatoid arthritis patients under infliximab treatment . OBJECTIVE : To determine whether anti- P01375 alpha ( infliximab ) treatment affects B cell activation in patients with rheumatoid arthritis ( RA ) METHODS : B cell activation was analyzed in fifteen anti- P01375 -treated RA patients . CD23 expression was used as a B cell activation marker and was studied before and after three months of infliximab treatment . PBMC were stimulated with anti-CD3 mAb during 18 h and were separated by rosseting into E+ and E-cells . B cells were assessed in E-population by double staining with P15391 and CD23 . ELISA assays were used to assess both soluble P01375 alpha and circulant immune complexes ( Q96RK0 ) containing P01375 alpha . We also used B cells from tonsils to establish the relationship between B cell activation and P01375 alpha Q96RK0 . RESULTS : The proportion of B cells expressing CD23 was higher before infliximab exposure than after treatment ( 48.3 +/- 16.7 versus 29.5 +/- 12.5 , p = 0.007 ) . T-B cell interactions were assessed by means of blocking antibodies to CD154 , P25942 , Q07108 , and P05107 ; these interactions were not specially affected by infliximab treatment . We could demonstrate Q96RK0 containing P01375 alpha after infliximab treatment , these Q96RK0 , similarly to others IgG-containing immune complexes , were capable to downregulate CD23 on B cells . CONCLUSIONS : DB00065 treatment in RA downregulates CD23 expression on T-cell activated B cells . This downregulation is connected with the presence of Q96RK0 containing P01375 alpha . Presumably , the Fc gamma RIIb1 endows IgG-containing immune complexes , as P01375 alpha-anti- P01375 alpha , with the capacity to regulate B cells and inflammatory cells . DB00065 treatment for resistant Achilles tendonitis . Undifferentiated spondyloarthropathies USPA can sometimes be refractory to usual disease modifying agents . Anti-tumor necrosis factor P01375 agents have been shown to be effective in spondyloarthropathies . Few articles described the efficacy of P01375 antagonists in USPA . Our patient had refractory Achilles tendonitis as an early manifestation of USPA which responded dramatically to infliximab treatment . DB00065 as disease-modifying therapy . Recent insights in the pathophysiology of Crohn 's disease have revealed that tumour necrosis factor ( P01375 ) plays a pivotal role in mucosal inflammation . DB00065 is a chimeric anti- P01375 monoclonal antibody with potent anti-inflammatory effects , probably based on apoptosis of inflammatory cells . Numerous controlled trials have demonstrated efficacy in both active and fistulating Crohn 's disease . Appropriate indications for using infliximab and growing experience with safety aspects have made this treatment a highly valuable tool in the management of Crohn 's disease . Mycobacterium marinum infection after infliximab therapy . A case of Mycobacterium Marinum infection of the nasal cavity is described . A 57 years old man was being on DB00065 for 2 years for severe psoriasis presented with five months history of epistaxis , nasal blockage and snoring . Local examination revealed bilateral nasal mass . The diagnosis of mycobacterial infection was suspected based upon the histopathological finding of granuloma in the biopsy specimen , and later confirmed by Mycobacterial culture . The patient was treated with 3 months therapy of Ethambutol and DB01045 with good clinical response.The clinical presentation of the case is discussed with a review of the literature about current guidelines for prophylaxis and other preventive strategy for infection among patients receiving P01375 antagonists . Treatment of intestinal Behçet 's syndrome with chimeric tumour necrosis factor alpha antibody . Few patients with Behçet 's syndrome have gastrointestinal ulceration . Such patients are difficult to treat and have a higher mortality . Faced with refractory symptoms in two patients with intestinal Behçet 's , we used the tumour necrosis factor alpha ( P01375 ) monoclonal antibody infliximab to induce remission . Both women ( one aged 27 years , the other 30 years ) presented with orogenital ulceration , pustular rash , abdominal pain , bloody diarrhoea due to colonic ulceration , weight loss , and synovitis . One had thrombophlebitis , digital vasculitis , perianal fistula , and paracolic abscess ; the other had conjunctivitis and an ulcer in the natal cleft . Treatment with prednisolone , methyl prednisolone , and thalidomide in one and prednisolone , colchicine , and cyclosporin in the other was ineffective . After full discussion , infliximab ( 3 mg/kg , dose reduced because of recent sepsis in one , and 5 mg/kg in the other ) was administered . Within 10 days the ulcers healed , with resolution of bloody diarrhoea and all extraintestinal manifestations . A second infusion of infliximab was necessary eight weeks later in one case , followed by sustained ( > 15 months ) remission on low dose thalidomide . Remission was initially sustained for 12 months in the other but thalidomide had to be stopped due to intolerance , and a good response to retreatment lasted only 12 weeks without immunosuppression , before a third infusion . The cause of Behçet 's syndrome is unknown but peripheral blood P08575 gammadelta T cells in Behçet 's produce > 50-fold more P01375 than controls when stimulated with phorbol myristate acetate and anti-CD3 . DB00065 could have a role for inducing remission in Behçet 's syndrome . Incidence and clinical significance of immunogenicity to infliximab in Crohn 's disease : a critical systematic review . BACKGROUND : DB00065 ( IFX ) is a chimeric ( mouse/human ) anti- P01375 monoclonal antibody approved for the treatment of refractory luminal and fistulizing Crohn 's disease ( CD ) . It is a source of potential immunogenicity for humans , with the occurrence of anti-infliximab antibodies ( ATIs ) , which are thought to interfere with the pharmacodynamics and/or pharmacokinetics of the compound . It remains unclear whether ATIs have any clinical importance for drug efficacy or safety . We review studies specifically evaluating the incidence of ATIs in CD and their impact on the efficacy and safety of IFX . METHODS : A systematic review was undertaken by electronic searches of the PubMed and SCOPUS databases from earliest records to October 2008 , as well as reference lists of all relevant articles and relevant abstracts from meetings . RESULTS : The biological and clinical mechanisms of ATI development are poorly understood . The incidence of ATIs in vivo depends on multiple analytical and clinical factors , both patient- and treatment-related . The presence of ATIs is weakly and variably associated with clinical response or infusion reactions , but not with reactions relevant to clinical decision-making . Enormous variation in the methods of reporting ATIs and immunogenicity of IFX make almost any interpretation possible from different studies , but few have clinical relevance . CONCLUSIONS : There is no clear evidence that ATIs have an impact on efficacy or safety , nor a need to measure or prevent them in clinical practice . Circulating drug concentration may be a more relevant measure of immunogenicity . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Effects of infliximab on cytokines , myeloperoxidase , and soluble adhesion molecules in patients with juvenile idiopathic arthritis . OBJECTIVE : DB00065 is effective and well tolerated in the treatment of juvenile idiopathic arthritis ( JIA ) . The aim of the present study was to measure circulating levels of inflammatory mediators in patients with JIA during treatment with infliximab . METHODS : Eight patients with active JIA refractory to standard treatments were treated with infliximab ( 3-4 mg/kg ) at weeks 0 , 2 and 6 and thereafter at approximately 6-week intervals up to 24 weeks . RESULTS : All patients ( n = 8 ) responded to the treatment . By 6 weeks of treatment the number of active joints had reduced from 16+/-4 ( mean+/-SEM ) to 4+/-1 ( p < 0.01 ) and P02741 ( CRP ) levels had fallen from 31+/-8 to 8+/-3 ( p < 0.001 ) . DB00065 treatment also reduced the serum concentrations of interleukin-6 ( P05231 ) , myeloperoxidase ( P05164 ) , and soluble adhesion molecules P05362 ( intercellular adhesion molecule-1 ) , and P16581 . Tumour necrosis factor-alpha ( TNFalpha ) levels tended to increase while the concentrations of endogenous P01375 antagonists ( sTNF-RI and sTNF-RII ) reduced in most patients during treatment . CONCLUSIONS : DB00065 reduced serum levels of P05231 , P05164 and soluble adhesion molecules in JIA patients , producing a good clinical response to the treatment . [ Efficacy and security of tumor necrosis factor antagonists in the treatment of rheumatoid arthritis ] . In the last decade , tumor necrosis factor ( P01375 ) antagonists had supposed an important therapeutic advance in the treatment of patients with rheumatoid arthritis ( RA ) in both early and established RA . Three agents currently available -- infliximab , etanercept , and adalimumab -- have been designed to modify the biologic effects of P01375 . DB00065 and adalimumab are monoclonal antibodies , whereas etanercept is a soluble protein , with different pharmacokinetic and pharmacodynamic properties , conditioning some possible adverse effects . Although comparative studies are not available , the 3 drugs have demonstrated efficacy and security , with a better quality of life of patients with RA . DB00065 , etanercept and adalimumab have been proved alone and in combination with methotrexate , with a better therapeutic , clinical , radiological and functional response in the group under combined therapy . Both clinical trials and post-market experience have demonstrated the security of these drugs , minimizing the risks with an adequate selection of patients . P04150 signaling in a bronchial epithelial cell line . Glucocorticoids are an effective anti-inflammatory therapy for the treatment of asthma . The anti-inflammatory effects of glucocorticoids may be due to the inhibition of transcription factors that regulate cytokine synthesis . Because of the potential role of the bronchial epithelium in asthmatic inflammation and the possibility that this cell may be the main target of inhaled glucocorticoids , we have characterized glucocorticoid receptors ( GR ) and GR signaling in the human bronchial epithelial cell line BEAS-2B . Western blot analysis and radioligand binding studies demonstrated that BEAS-2B cells have functional GR that bind to dexamethasone ( DB00514 ) ( dissociation constant = 5.6 nM and maximal density of binding sites = 228 +/- 3.3 fmol/mg protein ) . GR were activated by DB00514 as assessed using a glucocorticoid-responsive reporter plasmid . Transfection of BEAS-2B cells with an activator protein-1 ( AP-1 ) reporter construct followed by 12-O-tetradecanoylphorbol-13-acetate ( TPA ) treatment resulted in a fivefold induction of reporter gene activity . Transfection with a nuclear factor ( NF ) -kappa B reporter construct followed by tumor necrosis factor-alpha ( P01375 ) treatment resulted in a 10-fold induction of reporter gene activity . DB00514 ( 10(-7) M ) markedly repressed both the induced AP-1 and NF-kappa B activity . The GR antagonist DB00834 inhibited the repressive effect of DB00514 on P01375 -induced NF-kappa B activity by 81 % but only counteracted the repressive effect of DB00514 on TPA-induced AP-1 activity by 43 % . These studies demonstrate that cross-signaling between AP-1 and NF-kappa B with GR may explain the anti-inflammatory properties of glucocorticoids in airway epithelial cells . DB00065 for psoriasis . This review summarizes the use of inflximab in psoriasis and other immune-mediated inflammatory disorders ( IMIDs ) . The magnitude and speed of the response to infliximab monotherapy of moderate to severe psoriasis vulgaris is substantial , being similar to those achieved with cyclosporin . In contrast with cyclosporin , clinical improvement after the initial 3 intravenous influsions of infliximab is maintained for as long as 6 months in approximately half the patients with the absence of any additional treatment . Additionally , infliximab monotherapy normalizes keratinocyte proliferation and differentiation and markedly decreases epidermal inflammation . These results provide a convincing argument for the role of P01375 in the pathogenesis of psoriasis and for the clinical development of infliximab for the treatment of psoriasis .

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interacts_with DB00054?

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Q13444 is an adhesion receptor for platelet P08514 -IIIa and induces platelet activation . Cell adhesion and proteolytic matrix degradation are central processes in atherosclerosis . Being a member of the family of ADAMs ( " a disintegrin and metalloproteinase " ) , metargidin ( Q13444 ) combines a metalloproteinase domain and an RGD aminoacid sequence . We studied the potential role of Q13444 as an adhesion receptor on endothelial cells and interactions between platelets and Q13444 with respect to platelet adhesion , activation and thrombus formation . Q13444 was found to be expressed on cultured endothelial cells ( HUVEC ) . Platelet adhesion to immobilized recombinant Q13444 was effectively enhanced under both static and high shear rate conditions reaching the maximum level of adhesion to fibrinogen . Consistently , platelet adhesion onto Q13444 overexpressing endothelial cells was significantly increased . Adhesion to Q13444 was reduced by blockade of P08514 -IIIa using neutralizing anti-alpha(IIb)beta3 mAbs ( DB00054 , 2G12 ) , but not by anti-alpha(v)beta3 ( LM609 ) . Soluble Q13444 binds to activated but not to resting P08514 -IIIa . Moreover , platelets adherent to Q13444 additionally attracted platelets under high shear rates indicating an initial role of platelet- Q13444 interactions for thrombus formation . Furthermore , incubation of platelets with soluble Q13444 showed a dose-dependent increase in secretion of CD62P and P29965 . Q13444 is expressed on endothelial cells and can serve as an adhesion receptor for platelets via P08514 -IIIa binding . Platelet adhesion to Q13444 leads to platelet activation , secretion and promotes thrombus formation . Thus , Q13444 may represent a novel target for antithrombotic strategies in cardiovascular pathologies . Altered P25942 and P12830 expression -- putative role in oral lichen planus . BACKGROUND : Oral lichen planus ( OLP ) is characterized among other features by apoptosis of basal keratinocytes . To identify potential regulatory mechanisms associated with basal cell apoptosis in OLP , we investigated the expression of P25942 , P29965 ( P29965 ) , P16070 and epithelial ( E ) -cadherin . METHODS : Biopsies from 22 patients with OLP were investigated by immunohistochemistry for detection of P25942 , P29965 , P12830 , P16070 , Laminin-5 and Collagen IV , double-labelling for P25942 and CD3 , and in situ mRNA hybridization for P25942 and P29965 . RESULTS : In actively diseased areas of OLP lesions , basal keratinocytes did not express P25942 and were focally P12830 -negative , in contrast to non-diseased areas and normal oral mucosa . Demonstration of intraepithelial T cells expressing P25942 and P29965 , indicates a potential role in inflammatory cell responses involved in the disease process of OLP . CONCLUSION : T cells may orchestrate inflammatory cell responses in OLP via P25942 - P29965 interactions . As basal keratinocytes downregulate P25942 , they may escape P25942 - P29965 -induced apoptosis in OLP . On the other hand , loss of P12830 expression may contribute to epithelial basal cell destruction and T-cell migration into the epithelial compartment in OLP . Neurokinin-1 receptor antagonism in a rat model of subarachnoid hemorrhage : prevention of upregulation of contractile ETB and P28222 receptors and cerebral blood flow reduction . OBJECT : Cerebral vasospasm following subarachnoid hemorrhage ( Q53FZ2 ) leads to reduced cerebral blood flow ( Q03701 ) and to cerebral ischemia , in some cases even producing infarction and long-term disability . The goal of the present study was to investigate the hypothesis that inhibition of neurokinin-1 receptors ( NK1Rs ) by administration of L-822429 blunts the decrease in Q03701 as well as cerebrovascular receptor upregulation in an animal model of Q53FZ2 . METHODS : Subarachnoid hemorrhage was induced in rats by injection of 250 microl of blood into the prechiasmatic cistern . The P25103 inhibitor L-822429 was injected intracisternally 30 minutes and 24 hours after the induction of Q53FZ2 . Two days after Q53FZ2 induction , the basilar arteries were harvested , and contractile responses to endothelin-1 ( ET-I , an P25101 - and ETB-receptor agonist ) and 5-carboxamidotryptamine ( a 5-hydroxytryptamine- I1 [ 5-HT1 ] -receptor agonist ) were investigated using sensitive myographs . To determine whether Q8TDQ1 inhibition had an influence on local Q03701 after post- Q53FZ2 , a quantitative autoradiographic technique was used . After Q53FZ2 , the vascular receptor phenotype was changed in cerebral arteries through upregulation of contractile ET , and P28222 receptors , while regional and total Q03701 were markedly reduced . Treatment with the selective P25103 inhibitor L-822429 prevented both the receptor upregulation and the reduction in regional and global Q03701 . CONCLUSIONS : The data reveal the coregulation of vascular receptor changes and blood flow effects , and also show that interaction with a small-molecule P25103 antagonist is a promising area of focus for the development of specific treatments for Q53FZ2 . DB00054 pharmacodynamics are unaffected by antecedent therapy with other P08514 /IIIa antagonists in non-human primates . BACKGROUND : Tirofiban and eptifibatide are currently approved for the medical stabilization of non-ST segment elevation acute coronary syndromes . In patients undergoing percutaneous coronary intervention ( P05154 ) during infusion of these drugs , conversion to abciximab , which has long term proven clinical efficacy and cost-effectiveness , following P05154 may be desirable . The purpose of this study was to determine if the binding or pharmacodynamics of abciximab is affected by a prior infusion of either tirofiban or eptifibatide . METHODS : In vitro binding experiments were performed to determine if prior exposure to tirofiban or eptifibatide altered the affinity and extent of binding of abciximab to P08514 /IIIa . For in vivo experiments , cynomolgus monkeys were pretreated with a bolus and 18 hour infusion of saline , tirofiban , or eptifibatide . At the end of the initial treatment , a bolus and 12 hr infusion of abciximab was started without delay . Inhibition of platelet aggregation , P08514 /IIIa receptor blockade and abciximab pharmacokinetics were measured during and after both infusions . RESULTS : Equilibrium binding of abciximab in vitro was unaffected by tirofiban or eptifibatide . The extent and duration of abciximab inhibition of ex vivo platelet aggregation , receptor blockade , and abciximab pharmacokinetics in monkeys during and after the abciximab infusion were not affected by prior infusion of the animals with tirofiban or eptifibatide . CONCLUSIONS : In vitro and in vivo studies revealed that the molecular interaction of abciximab with the platelet P08514 /IIIa receptor is not altered by immediate prior exposure of platelets to small molecule P08514 /IIIa antagonists . These preclinical studies suggest that the efficacy of abciximab should not be impaired if it is initiated following termination of therapy with small molecule P08514 /IIIa antagonists . Concentration-dependent effect of abciximab on platelets and neutrophils in a model of cardiopulmonary bypass . DB00054 is a P08514 /IIIa antagonist used in percutaneous coronary interventions to avoid platelet activation , thrombosis and inflammation . We investigated whether abciximab influenced platelet activation and platelet interaction with neutrophils and polyvinyl chloride ( PVC ) in a cardiopulmonary bypass ( P15086 ) model . Isolated platelets , preincubated with and without 0.1-20 microg/mL of abciximab , were resuspended with neutrophils in plasma and recirculated by a roller pump . Platelet , but not neutrophil adhesion to PVC was inhibited by abciximab . Only high doses of abciximab reduced platelet aggregation size , but simultaneously increased platelet-neutrophil aggregation . DB00054 had no effect on platelet CD62P expression or degranulation , but platelet activation on platelet-neutrophil aggregates increased with high doses . Only low doses inhibited neutrophil degranulation . The concentration-dependent effect of abciximab on platelet-neutrophil interaction reduces its usefulness and stresses the dependency on experimental design in the evaluation of abciximab . Our study does not support the use of abciximab alone in P15086 . However , incorporation of surface-coating the biomaterial with abciximab may be an interesting option . Potential future clinical applications for the P08514 /IIIa antagonist , abciximab in thrombosis , vascular and oncological indications . DB00054 ( ReoPro ) is a mouse-human chimeric monoclonal antibody Fab fragment of the parent murine monoclonal antibody DB00054 , and was the first of these agents approved for use as adjunct therapy for the prevention of cardiac ischemic complications in patients undergoing percutaneous coronary intervention ( P05154 ) . DB00054 binds with high avidity to both the non-activated and activated form of the P08514 /IIIa receptor of platelets , the major adhesion receptor involved in aggregation . Additional cardiovascular indications for abciximab are unstable angina , carotid stenting , ischemic stroke and peripheral vascular diseases . DB00054 also interacts with two other integrin receptors ; the a av b b3 receptor , which is present in low numbers on platelets but in high density on activated endothelial and smooth muscle cells , and a aMb b2 integrin which is present on activated leukocytes . Cell types that express integrins P08514 /IIIa and a av b b3 such as platelets , endothelial and tumor cells have been implicated in angiogenesis , tumor growth and metastasis . Since abciximab interacts with high avidity to integrins P08514 /IIIa and a av b b3 , it is reasonable to assume that it may possess anti-angiogenic properties in angiogenesis-related diseases , as well as anti-metastastatic properties in case of disseminating tumors expressing the target integrin receptors . Platelet-derived microparticle formation involves glycoprotein IIb-IIIa . Inhibition by RGDS and a Glanzmann 's thrombasthenia defect . While the physiologic role of platelet microparticles may include a stable , physical dispersion of concentrated surface procoagulant activity the mechanism(s) of platelet vesiculation remains unknown . We demonstrate using flow cytometric methods a central role for the beta 3 integrin glycoprotein ( GP ) IIb-IIIa complex and its ligand tetrapeptide DB00125 - DB00145 - DB00128 - DB00133 ( RGDS ) binding site in platelet vesiculation . Time- and calcium-dependent vesiculation of platelets in response to ADP , collagen , thrombin , phorbol myristate acetate , and the thrombin peptide SFLLRN were dramatically inhibited , in a concentration-dependent manner , by monoclonal antibodies to P08514 -IIIa ( A2A9 , DB00054 , PAC1 ) and RGDS . Complete inhibition with A2A9 and RGDS occurred at 7.5 micrograms/ml and 75 microM , respectively , while control antibodies and a mock peptide had no effect . Platelet vesiculation requires intact P08514 -IIIa and is fully supported by the intracellular pool of P08514 -IIIa alone since de-complexing of this heterodimer by calcium chelation completely abolished microparticle formation in response to collagen ( no alpha-granule release ) but not to thrombin or SFLLRN . A central role for P08514 -IIIa is supported by the near total inability of Glanzmann 's thrombasthenic ( type I ) platelets to vesiculate in response to thrombin , ADP , collagen , and phorbol 12-myristate 13-acetate . This extends the biologic roles of P08514 -IIIa to include platelet vesiculation and suggests that one or all of its binding ligands play a role . Arterial reocclusion and persistent distal occlusion after thrombus aspiration . BACKGROUND AND PURPOSE : Early reocclusion of intracranial arteries can lead to poor clinical outcome . We report reocclusion detection after endovascular clot aspiration , followed by administration of P08514 -IIIa antagonist under continuous ultrasound monitoring . CASE DESCRIPTION : A 73-year-old man developed the right middle cerebral artery ( MCA ) occlusion with NIHSS 17 points , 6 days after aortic valve replacement . Recanalization was achieved with Penumbra™ system and reocclusion was detected with transcranial Doppler ( P24386 ) 30 minutes postcompletion of intra-arterial procedure . Proximal recanalization was achieved with the second thrombus aspiration while M2 MCA occlusion persisted beyond the reach of the device . Intravenous abciximab was administered under continuous P24386 monitoring . Recanalization with Thrombolysis in Brain Ischemia ( TIBI ) flow grade 4 was observed at 60 minutes postintervention accompanied with clinical recovery to NIHSS 3 points . DB00054 was given for 12 hours with no hemorrhagic transformation on repeat CT scan . Patient was discharged home with mild left pronator drift and facial droop , and his modified ranking score was 1 at 6-week follow-up visit . CONCLUSIONS : Early arterial reocclusion can occur after successful thrombus aspiration while P08514 -IIIa antagonist administration may lead to subsequent recanalization of persisting distal occlusions not amenable to mechanical removal . The cytokines ( P01579 , P60568 , P05112 , P22301 , Q16552 ) and Treg cytokine ( TGF-beta1 ) levels in adults with immune thrombocytopenia . Previous studies have indicated that autoimmune diseases might be caused by an imbalance of T helper cells ( Th ) , cytokines , and regulatory T cells ( Treg ) cytokines . We measured the plasma concentrations of Th1-associated cytokines ( P01579 , P60568 ) , Th2 -associated cytokines ( P05112 , P22301 ) , Th17-associated cytokine ( Q16552 ) and Treg -associated cytokine ( TGF-beta1 ) in adult patients with immune thrombocytopenia ( ITP ) and evaluated their clinical relevance . Plasma P01579 , P60568 , P05112 , P22301 , Q16552 and TGF-beta1 concentrations of 52 ITP patients and 30 age- and sex-matched healthy controls were measured by enzyme-linked immunosorbent assay method ( ELISA ) . Concentration of Th2 cytokines ( P05112 and P22301 ) were significantly higher in ITP patients compared to controls ( P < 0.05 ) . However , concentrations of Th1 cytokines ( P01579 , P60568 ) , Th17 cytokine ( Q16552 ) and Treg cytokine ( TGF-beta1 ) were lower in ITP patients ( P < 0.05 ) . Concentration of Q16552 was significantly higher in chronic ITP patients compared to severe ITP patients ( P < 0.05 ) , and no significant difference of cytokine concentration among the other subgroups in ITP patients was found . Among the ITP patients , concentration of P01579 correlated positively and significantly with PAIgG ( r = 0.48 , P = 0.02 ) . A significant correlation was neither found between other cytokine levels and platelet count , nor between cytokine levels and megakaryocytes number , nor between cytokines levels and PAIgG or P08514 /IIIa and/or GPIb/IX autoantibodies . The present study demonstrates that an imbalance of Th and Treg cytokines may mediate the pathogenesis of ITP . Investigation of interaction of human platelet membrane components with anticoagulant drugs DB00054 and DB00063 . DB00054 ( Abci ) and eptifibatide ( Epti ) are antiaggregate drugs which may reduce thrombotic complications in acute coronary syndromes . The aim of this work was the investigation of the interaction between the phospholipid- P08514 /IIIa glycoprotein complex and Abci or Epti , and the influence of these drugs on the phospholipid ratio in the platelet membrane . The interaction between the phospholipid- P08514 /IIIa glycoprotein complex and antiaggregate drugs were investigated using the Surface Plasmon Resonance Imaging technique ( SPRI ) . Phospholipids phosphatidylinositol ( PI ) , phosphatidylserine ( PS ) , phosphatidylethanolamine ( PE ) , phosphatidylcholine ( PC ) and sphingomyelin ( SM ) were first immobilized onto the gold chip surface . The phospholipid ratio in the platelet membrane was determined by the HPLC . Only PI , PS , PE and PC were determined . Human platelets treated ' in vitro ' with Abci or Epti exhibit changes in the phospholipid ratio in the platelet membrane . The ratio of PS decreases and PC rises . The SPRI distinctly shows interactions between phospholipids and glycoprotein P08514 /IIIa , and between the phospholipid-glycoprotein P08514 /IIIa complex and Abci or Epti . The interaction between phospholipids and glycoprotein P08514 /IIIa is growing in the sequence : PI << SM < PE < PC < PS . The interaction between phospholipid-glycoprotein P08514 /IIIa complex and Abci/Epti is growing in the sequence : PS < PI < PC < PE < SM . SPRI was proved to be excellent tool for observation of such interactions . The disulfide-rich region of platelet glycoprotein ( GP ) IIIa contains hydrophilic peptide sequences that bind anti- P05106 autoantibodies from patients with immune thrombocytopenic purpura ( ITP ) . Immune thrombocytopenic purpura ( ITP ) is an autoimmune blood disease caused by autoantibody-mediated destruction of blood platelets . Platelet glycoprotein ( GP ) IIb/IIIa is a common target for antiplatelet autoantibodies . The present studies were undertaken (1). to confirm whether the disulfide rich repeat region of P05106 contains target epitopes for antiplatelet antibodies in patients with ITP ; (2). to determine whether these antigens were defined by peptide sequences in the absence of post-translational modification ; and (3). to correlate observed immunologic reactivity with the recently solved X-ray crystallographic structure of an analogous integrin complex , the vitronectin receptor , alpha(V)beta(3) . Recombinant fusion proteins of four P05106 extracellular sequences were prepared and purified . Immunoblotting results with purified recombinant peptides showed potent reactivity of 16 of 24 ITP patient serum anti- P08514 /IIIa antibodies with the fusion protein containing the P05106 sequence of residues from 468 to 691 . These results are consistent with a report by Kekomaki et al. that a 50 kDa chymotryptic digestion product of P05106 isolated from blood platelets contains target epitopes for serum antiplatelet antibodies in 16 of 33 ITP patients . Smaller peptides including residues 446-501 and residues 593-691 each reacted with only 5 of the 24 patient sera ; furthermore all but 3 of these interactions were very weak . Visualization of the conformation of the extracellular portion of alpha(V)beta(3) reveals the location of the 222-residue antigenic P05106 ( beta(3) ) peptide ' B ' at the immediately extracellular region of the protein that includes a beta-tail domain and several integrin- P01133 domains . In summary , predictions of hydrophilicity , surface accessibility and antigenicity and the three dimensional structure of the beta(3) integrin correlate with autoantibody binding to a recombinant P05106 peptide ' B ' containing residues 468-691 . Antithrombotic therapy in the acute phase : new approaches . Both anticoagulants and antiplatelet agents have been advocated , used and studied for the treatment of acute ischemic stroke . Randomized trials of unfractionated heparin , low-molecular-weight heparin and heparinoids have failed to show an overall benefit to these agents largely because the benefits in reducing thromboembolic events are offset by the increased risk of bleeding complications . The International Stroke Trial , the Trial of ORG 10172 Acute Stroke Treatment and studies of fraxaripine all failed to show an overall benefit to anticoagulation in the patients studied . DB00945 has been shown to offer a modest benefit when studied in patients treated within 48 h of stroke onset . DB05099 is an antithrombotic agent that acts by reducing circulating fibrinogen levels . Patients treated within 3 h of stroke symptom onset had a better functional outcome at 90 days compared to placebo-treated patients with both the benefits and the risk of intracerebral bleeding related to the fibrinogen lowering achieved . DB00054 is a blocker of the platelet P08514 /IIIa receptor . A dose finding safety study suggests that in doses up to that typically given in patients with acute coronary occlusion syndromes , there is no increased risk of symptomatic intracerebral bleeding and suggestions of potential benefits on neurological outcome . Neonatal platelets are less reactive than adult platelets to physiological agonists in whole blood . Previous studies have reported that the platelets of healthy term neonates have either diminished or normal reactivity compared to the platelets of adults . To circumvent the methodologic problems of previous studies , we used a whole blood flow cytometric method to study neonatal platelet reactivity to thrombin , a combination of ADP and epinephrine , and U46619 ( a stable thromboxane A2 analogue ) . Inclusion in the assay of the peptide GPRP ( an inhibitor of fibrin polymerization ) enabled us to study platelet reactivity to human alpha-thrombin in whole blood . Umbilical cord blood and day 1 peripheral blood were collected from 30 healthy term neonates and compared to peripheral blood from 20 normal adults . In whole blood samples without added agonist , there were no significant differences between neonates and adults in the platelet binding of monoclonal antibodies 6D1 ( GPIb-specific ) or DB00054 ( P08514 -IIIa complex-specific ) . As determined by P28222 ( a P16109 -specific monoclonal antibody ) , neither neonates nor adults had circulating degranulated platelets . However , in both cord and peripheral whole blood samples , neonatal platelets were significantly less reactive than adult platelets to thrombin , ADP/epinephrine , and U46619 , as determined by the extent of increase in the platelet surface expression of P16109 and the P08514 -IIIa complex , and the extent of decrease in the platelet surface expression of the GPIb-IX complex. ( ABSTRACT TRUNCATED AT 250 WORDS ) Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Time course of the effects of a single bolus injection of F(ab')2 fragments of the antiplatelet P08514 /IIIa antibody 7E3 on arterial eversion graft occlusion , platelet aggregation , and bleeding time in dogs . The time course of the effects of a single intravenous bolus injection of 10 mg/kg aspirin or 0.8 mg/kg F(ab')2 fragments of the monoclonal antiplatelet glycoprotein IIb/IIIa receptor antibody DB00054 [ 7E3-F(ab')2 ] on arterial occlusion , platelet aggregation , and bleeding time was studied in 30 dogs with an everted ( inside out ) carotid arterial segment inserted into the femoral artery . In the absence of an antiplatelet agent , the eversion grafts occluded spontaneously with platelet-rich thrombus within 30 minutes . With aspirin , arterial occlusion persisting for 2 hours occurred in 5 of 10 dogs and cyclic occlusion and reflow in 4 animals ; arterial occlusion was observed in all dogs at 24 hours . With 7E3-F(ab')2 , arterial patency persisted throughout a 2-hour observation period in all of 10 dogs and for 24 hours in 4 of the 10 dogs . Contralateral eversion grafting 24 hours after aspirin or 7E3-F(ab')2 injection was associated with graft patency for 2 hours in 1 of 5 aspirin dogs and in 3 of 5 7E3-F(ab')2 dogs ; patency persisted for 24 hours . In dogs grafted 48 hours after aspirin or 7E3-F(ab')2 injection , patency at 24 hours was seen in 0 of 5 dogs given aspirin and 3 of 5 dogs given 7E3-F(ab')2 . The overall frequencies of arterial graft patency at 2 , 24 , 48 , and 72 hours after study drug injection were significantly higher in the 7E3-F(ab')2 groups than in the aspirin groups ( P < .0005 , n = 10 in each group ; P < .05 , n = 15 ; P < .005 , n = 15 ; and P = .05 , n = 5 , respectively ) . ( ABSTRACT TRUNCATED AT 250 WORDS ) Characterization of canine platelet P16109 ( CD 62 ) and its utility in flow cytometry platelet studies . 1. P16109 ( P16109 , GMP140 , or CD62 ) a member of lectin-like adhesive proteins is expressed on the surface of activated degranulated canine platelets and is the calcium-dependent receptor for leukocyte adhesion . 2 . The electrophoretic mobility of P16109 , by Western blot analysis and immunoprecipitation from radiolabeled membranes of canine and human platelets , was similar or identical and immunocytochemical studies localized P16109 in internal vesicles similar to the alpha granule localization in human platelets . 3 . Two antibodies to human P16109 KC4.1 and Q99440 .2 crossreacted with canine platelets whose surface binding , in response to agonists thrombin , calcium ionophore ( A23187 ) , phorbol esters and ADP , was similar . 4 . Anti- P16109 antibodies in conjunction with crossreacting anti- P08514 /IIIa antibodies ( A2A9 , DB00054 , RUU-PL7F12 ) enables the analysis of activated platelets , platelet-derived microparticles and platelet-leukocyte interactions in canine models by flow cytometry . Genetic ablation of Adamts13 gene dramatically accelerates the formation of early atherosclerosis in a murine model . OBJECTIVE : Q76LX8 ( a disintegrin and metalloprotease with thrombospondin type 1 repeats-13 ) cleaves P04275 , thereby modulating thrombosis and inflammation . Low plasma Q76LX8 activity is associated with cardiovascular events , including myocardial and cerebral infarction . Here , we investigated the role of Q76LX8 in the development of early atherosclerosis in a murine model . METHODS AND RESULTS : P02649 -null ( ApoE(-/-) ) and Adamts13-null ( Adamts13(-/-) ) ApoE(-/-) mice were fed with a high-fat Western diet for 12 weeks . Atherosclerotic lesions in the aorta and aortic roots were quantified after staining . Leukocyte rolling and adhesion onto cremaster venules after oxidative injury were determined by intravital microscopy . Although plasma cholesterol levels were largely similar in both groups , the extent of atherosclerotic lesions in the aorta en face and in the aortic roots in the Adamts13(-/-)ApoE(-/-) mice increased ≈ 5.5-fold ( P=0.0017 ) and ≈ 6.1-fold ( P=0.0037 ) , respectively . In addition , the ratio of plasma high- to low-molecular-weight P04275 multimers increased ≈ 3-fold . The leukocyte rolling velocities were significantly reduced ( P < 0.001 ) , with an increased number of leukocyte rolling ( P=0.0026 ) and macrophage infiltration into the atherosclerotic lesions in the Adamts13(-/-)ApoE(-/-) mice . CONCLUSIONS : Our results suggest that Q76LX8 plays a critical role in modulating the development of early atherosclerosis , likely through the proteolytic cleavage of ultra-large P04275 multimers , thereby inhibiting platelet deposition and inflammation . A new short chain RGD-containing disintegrin , accutin , inhibits the common pathway of human platelet aggregation . A new short-chain disintegrin , accutin , was purified from the Formosan Agkistrodon acutus venom by using of gel filtration , ion exchanger and reverse phase HPLC . The homogeneous protein is a 47-residue polypeptide with a molecular mass of 5241 Da containing an DB00125 - DB00145 - DB00128 sequence and seven cysteinyl residues at positions highly homologous to other disintegrins . Accutin dose-dependently inhibited human platelet aggregation stimulated by ADP , collagen , thrombin or the thromboxane analogue U46619 in platelet suspension with IC50 values of 66-267 nM . It was also active in inhibiting platelet aggregation of platelet-rich plasma . However , accutin apparently did not affect the shape change caused by these agonists . Accutin also inhibited fibrinogen-induced aggregation of human elastase-treated platelets in a dose-dependent manner . Furthermore , accutin dose-dependently inhibited the binding reaction of fluorescein isothiocyanate ( FITC ) -conjugated arietin , a member of the disintegrin family , to human platelets . In addition , the binding of FITC-conjugated accutin to platelets was almost completely blocked by a monoclonal antibody , DB00054 , raised against the platelet glycoprotein IIb/IIIa complex . On the other hand , accutin as well as other disintegrins , rhodostomin and arietin , exhibited an inhibitory effect on 7E3 binding toward platelets and endothelial cells in a dose-dependent manner . It is concluded that accutin , a new platelet aggregation inhibitor belonging to the short-chain disintegrin family , acts specifically on a binding epitope of P08514 /IIIa overlapping with that of DB00054 , leading to the blockade of fibrinogen binding to its receptor . DB01109 potentiation of collagen-induced platelet aggregation is related to the P08514 / P05106 receptor and not to the GPIb receptor , as tested by whole blood aggregometry . To determine whether heparin potentiation of platelet aggregation is related to platelet GP IIb/IIIa and GP Ib receptors , four series of experiments were performed on blood from normal volunteers . In the first experiment pretreatment with the monoclonal antibody DB00054 ( MAb DB00054 ) , which antagonizes at the GP IIb/IIIa receptor , potently inhibited the collagen-induced platelet aggregation ( p less than 0.001 ) . With heparin added to blood pretreated with MAb DB00054 , the aggregation increased ( p less than 0.005 ) to an extent similar to that when only saline was used for pretreatment . In the second experiment , monoclonal antibody 10E5 ( MAb 10E5 ) and peptide RGDS , substances which also antagonize at the GP IIb/IIIa receptor , decreased collagen-induced platelet aggregation to an extent similar to that after pretreatment with MAb DB00054 . Following pretreatment with RGDS , heparin increased platelet aggregation ( p less than 0.03 ) , while after pretreatment with antibody MAb 10E5 heparin did not enhance platelet aggregation . In the third experiment aurin , an inhibitor of P04275 and its interaction with the platelet GPIb receptor , decreased platelet aggregation dose-dependently . In the fourth experiment heparin enhanced platelet aggregation to a similar extent ( p less than 0.005 ) , regardless of pretreatment of the blood with saline , aurin or monoclonal antibody 6D1 ( MAb 6D1 ) , the latter an antagonist at the GP Ib receptor . In conclusion , the potentiation of collagen-induced platelet aggregation by heparin was not inhibited by MAb DB00054 , RGDS , aurin or MAb 6D1 , but was abolished by MAb 10E5 , implying that the heparin effect is related to activation of the platelet GP IIb/IIIa receptor complex . Minimal influence of tocilizumab on P01579 synthesis by tuberculosis antigens . P01579 ( P01579 ) production is a critical step of antituberculosis ( anti-TB ) immune response . The purpose of this study was to determine the influences of biologics , including the interleukin ( IL ) -6 receptor-inhibitor tocilizumab ( TCZ ) , and tumor necrosis factor ( P01375 ) antagonists infliximab ( P27352 ) and etanercept ( P25101 ) , on Mycobacterium tuberculosis ( MTB ) antigen-induced P01579 production . MTB antigen ( ESAT-6 and P27918 -10 ) -induced P01579 -releasing assay was performed with or without addition of biologics ( TCZ , P25101 , and P27352 ) using whole blood from patients with active TB . P25101 and P27352 inhibited P01579 production in a dose-dependent manner . In whole blood from TB patients , ESAT-6 stimulated significant production of P01579 ( 1.30 +/- 1.95 IU/ml ) , and TCZ did not inhibit P01579 production ( 1.56 +/- 1.88 IU/ml ) . P01579 production by ESAT-6 was inhibited by P25101 and P27352 ( 0.98 +/- 1.74 , 0.75 +/- 1.66 IU/ml , respectively ) . P27918 -10 stimulated significant production of P01579 ( 1.46 +/- 1.60 IU/ml ) , and TCZ did not inhibit P01579 production ( 1.51 +/- 1.77 IU/ml ) . P01579 production by P27918 -10 was inhibited by P25101 and P27352 ( 0.91 +/- 0.99 , 0.72 +/- 0.88 IU/ml , respectively ) . P24386 did not inhibit MTB antigen-induced P01579 production . As P01579 production is important in antimycobacterial host defenses , the minimal influence of TCZ on P01579 -releasing assay suggests a low risk of latent TB infection reactivation during tocilizumab therapy . [ P08514 -IIIa inhibitors ] . Therapy involving the use of anti- P08514 -IIIa inhibitors has progressively evolved in recent years for patients undergoing percutaneous coronary intervention or with acute coronary syndromes . Patients receiving anti-GP IIb-IIIa therapy have a lower risk of death or myocardial infarction than those receiving the classic anti-agregant , aspirin , alone . Two classes of products have been used in clinic , the chimeric monoclonal antibody Fab fragment , c7E3 or abciximab ( ReoPro ) , which has been the pioneer , and synthetic peptides or peptidomimetics such as eptifibatide ( Integrilin ) or tirofiban ( Agrastat ) . DB00054 is a long-acting , high-affinity receptor blocker , whereas eptifibatide and tirofiban have much shorter biological half-lives . Another property that differentiates these compounds is that the peptides bind exclusively to GP IIb-IIIa whereas c7E3 also binds to alpha v beta 3 , the vitronectin receptor . The potent inhibitory effect of these compounds increases the risk of bleeding . By carefully controlling the levels of heparin and by removing the sheath as early as possible , the hemorrhagic problems may be limited . Another potential complication is the rapid development of thrombocytopenia . The cause has yet to be found and for c7E3 no correlation with the development of HACA ( human anti-chimeric antibodies ) has been observed . Because of the chronic nature of coronary artery disease , evaluation of the readministration of c7E3 to the same patient two or even more times is under investigation . The first results do not show major problems . The best biological way to investigate the efficiency of anti- P08514 -IIIa has to be determined . Interestingly , a new point-of-care test has been proposed , while monoclonal antibodies are available that differentiate between nonoccupied and occupied P08514 -IIIa complexes . New antithrombotics for the treatment of acute and chronic arterial ischemia . The established antithrombotic agents are effective but they have limitations which have provided opportunities for the development of new antithrombotic compounds . Of these new agents , the antithrombin III-independent thrombin inhibitors and the platelet P08514 /IIIa receptor antagonists are the most advanced in their development . Other new antithrombotic agents include the antithrombin III-independent factor Xa inhibitors , activated protein C , soluble thrombomodulin and tissue factor pathway inhibitor . Of the P08514 /IIIa antagonists , the humanized DB00054 and integrin have been evaluated in phase III studies . The DB00054 was effective in preventing both short-term and longer-term complications of coronary angioplasty . The antithrombin III-independent thrombin inhibitors hirudin and hirulog have also been evaluated in phase III studies . The studies with hirudin as an adjuvant to coronary thrombolysis had to be terminated and restarted at lower dosages because of an unacceptable incidence at intracranial hemorrhage and the study with hirulog produced equivocal results . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . Neutrophil P16109 -glycoprotein-ligand-1 binding to platelet P16109 enhances metalloproteinase 2 secretion and platelet-neutrophil aggregation . Platelets and neutrophils constitute a high source of metalloproteinases ( MMPs ) , and their interactions via P16109 and P16109 -glycoprotein-ligand-1 ( Q14242 ) are involved in thrombosis , vascular remodelling , and restenosis . We investigated the impact of these interactions on platelet P08253 secretion and function in platelet and neutrophil aggregation . The secretion of P08253 from human platelets was significantly increased three-fold after thrombin activation , and enhanced two-fold in the presence of neutrophils . Neutrophil supernatant had no effect on platelet P08253 secretion . While no P08253 was detected in the supernatant of neutrophils , a high amount of P14780 was released by neutrophils , and remained unchanged upon thrombin activation or in the presence of platelets . Platelet P16109 , which increased significantly after activation , triggered platelet binding to neutrophils that was completely inhibited by P16109 or Q14242 antagonists , and was reduced by 50 % with a P08514 / IIIa antagonist . P16109 or Q14242 antagonism abolished the enhanced secretion of platelet P08253 in the presence of neutrophils and reduced platelet-neutrophil aggregation . Platelet activation and binding to neutrophils enhance the secretion of platelet P08253 via an adhesive interaction between P16109 and Q14242 , which contribute to increase platelet-neutrophil aggregation . Progress in the field of P08514 /IIIa antagonists . Platelet aggregation plays an important role in pathological situations such as myocardial infarction , unstable angina , peripheral artery disease , and stroke . Thus , pharmacological agents that specifically inhibit platelet aggregation are of great interest in the treatment and prevention of these cardiovascular diseases . Since binding of activated glycoprotein IIb/IIIa complex , a platelet surface integrin , to fibrinogen is the final step leading to platelet aggregation regardless of the initial stimulus , many researches have focused on the development of drugs that could antagonize this integrin . Three intravenous glycoprotein IIb/IIIa antagonists are currently marketed for the prevention of myocardial infarction in patients undergoing percutaneous intervention : DB00054 , DB00063 and Tirofiban . To further test the clinical efficacy of these agents , oral glycoprotein IIb/IIIa antagonists have been developed but only led to disappointing clinical results . Nevertheless , due to recognized usefulness of oral agents for the prevention and treatment of cardiovascular diseases , a great number of new orally active compounds are under clinical or preclinical evaluation . The aim of this review is to describe the chemical , pharmacological and clinical properties of existing and forthcoming glycoprotein IIb/IIIa antagonists . [ DB00054 ( ReoPro ) in the treatment of acute coronary syndromes ] . Platelet activation plays a major role in the pathophysiology of acute coronary syndromes ( ACS ) . Inhibition of platelet function is the basic pharmacological treatment of ACS . P08514 /IIIa inhibitors , a new class of potent antiplatelet agents , have been used in the treatment of ACS and in the prevention of complications after percutaneous coronary interventions ( P05154 ) . Several large clinical trials have demonstrated the effectiveness of this class of agents . The first of these agents to show beneficial effects after coronary interventions was the mouse/human chimeric Fab fragment antibody c7E3 abciximab ( ReoPro ) . The purpose of this article is to describe the pharmacology of abciximab and to review the results of the clinical trials carried out with the drug in patients with ACS , treated either with or without acute/elective P05154 . Assembly of a three-dimensional multitype bronchiole coculture model using magnetic levitation . A longstanding goal in biomedical research has been to create organotypic cocultures that faithfully represent native tissue environments . There is presently great interest in representative culture models of the lung , which is a particularly challenging tissue to recreate in vitro . This study used magnetic levitation in conjunction with magnetic nanoparticles as a means of creating an organized three-dimensional ( 3D ) coculture of the bronchiole that sequentially layers cells in a manner similar to native tissue architecture . The 3D coculture model was assembled from four human cell types in the bronchiole : endothelial cells , smooth muscle cells ( SMCs ) , fibroblasts , and epithelial cells ( EpiCs ) . This study represents the first effort to combine these particular cell types into an organized bronchiole coculture . These cell layers were first cultured in 3D by magnetic levitation , and then manipulated into contact with a custom-made magnetic pen , and again cultured for 48 h . Hematoxylin and eosin staining of the resulting coculture showed four distinct layers within the 3D coculture . Immunohistochemistry confirmed the phenotype of each of the four cell types and showed organized extracellular matrix formation , particularly , with collagen type I . Positive stains for CD31 , P04275 , smooth muscle α-actin , vimentin , and fibronectin demonstrate the maintenance of the phenotype for endothelial cells , SMCs , and fibroblasts . Positive stains for mucin-5AC , cytokeratin , and P12830 after 7 days with and without 1 % fetal bovine serum showed that EpiCs maintained the phenotype and function . This study validates magnetic levitation as a method for the rapid creation of organized 3D cocultures that maintain the phenotype and induce extracellular matrix formation . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . 7E3 F(ab')2 , an effective antagonist of rat alphaIIbbeta3 and alphavbeta3 , blocks in vivo thrombus formation and in vitro angiogenesis . DB00054 ( c7E3 Fab , ReoPro ) blocks P08514 /IIIa and alphavbeta3 and inhibits thrombotic and proliferative events only in humans and non-human primates . The bivalent F(ab')2 fragment is an effective anti-thrombotic agent in canine models . In the present study , 7E3 F(ab')2 was also found to bind to rat P08514 /IIIa ( KD = 27 +/- 4 microg/mL ) and alphavbeta3 ( KD = 9 +/- 8 microg/mL ) , to block in vitro rat platelet aggregation ( IC50 = 16 +/- 6 microg/mL ) , and to inhibit alphavbeta3-mediated microvessel sprout formation in a rat aortic ring assay . Following administration of 7E3 F(ab')2 ( 4 mg/kg ) to rats , platelet aggregation was completely blocked for up to 6 h and thrombus formation in response to a rat abdominal aorta double crush injury was prevented . Effective chronic dosing was achieved with 6 mg/kg daily I.P. injections . In vitro mixing experiments indicated that 7E3 F(ab')2 redistributed to unlabeled platelets in 2 h . Ex vivo , 7E3 F(ab')2 was detected on platelets for up to 4 days after a single 4-mg/kg injection . These data suggest that 7E3 F(ab')2 may be a useful agent to study the effects of P08514 /IIIa and alphavbeta3 blockade in rat models of thrombosis and vascular disease . Platelets and their chemokines in atherosclerosis-clinical applications . The concept of platelets as important players in the process of atherogenesis has become increasingly accepted due to accumulating experimental and clinical evidence . Despite the progress in understanding the molecular details of atherosclerosis , particularly by using animal models , the inflammatory and thrombotic roles of activated platelet s especially in the human system remain difficult to dissect , as often only the complications of atherosclerosis , i.e. , stroke and myocardial infarction are definable but not the plaque burden . Platelet indices including platelet count and mean platelet volume ( MPV ) and soluble mediators released by activated platelets are associated with atherosclerosis . The chemokine P02776 has multiple atherogenic activities , e.g. , altering the differentiation of T cells and macrophages by inhibiting neutrophil and monocyte apoptosis and by increasing the uptake of oxLDL and synergizing with P13501 . P13501 is released and deposited on endothelium by activated platelets thereby triggering atherogenic monocyte recruitment , which can be attenuated by blocking the corresponding chemokine receptor P51681 . Atheroprotective and plaque stabilizing properties are attributed to P48061 , which plays an important role in regenerative processes by attracting progenitor cells . Its release from luminal attached platelets accelerates endothelial healing after injury . Platelet surface molecules P08514 /IIIa , GP1bα , P16109 , Q9Y624 and the P25942 / P29965 dyade are crucially involved in the interaction with endothelial cells , leukocytes and matrix molecules affecting atherogenesis . Beyond the effects on the arterial inflammatory infiltrate , platelets affect cholesterol metabolism by binding , modifying and endocytosing LDL particles via their scavenger receptors and contribute to the formation of lipid laden macrophages . Current medical therapies for the prevention of atherosclerotic therapies enable the elucidation of mechanisms linking platelets to inflammation and atherosclerosis . Absence of potentiation with murine antiplatelet P08514 /IIIa antibody of thrombolysis with recombinant tissue-type plasminogen activator ( rt-PA ) in a canine venous thrombosis model . F(ab')2 fragments of a murine monoclonal anti-platelet P08514 /IIIa antibody ( DB00054 ) are a potent platelet aggregation inhibitor , which in a canine coronary artery thrombosis model accelerate lysis with recombinant tissue-type plasminogen activator ( rt-PA ) and prevent reocclusion ( 7 ) . In the present study , we have investigated the potential value of platelet aggregation inhibition as adjunctive therapy to lysis of venous thrombi , by measuring the thrombolytic potency of 7E3-F(ab')2 and rt-PA used alone or in combination , in dogs with a 125I-fibrin labeled femoral vein thrombus . The dose-response of thrombolysis with rt-PA infused over 4 hours was linear : doses of 0.075 mg/kg , 0.15 mg/kg and 0.3 mg/kg produced 37 +/- 3 , 57 +/- 11 and 83 +/- 4 % lysis respectively , against a background value of 20 +/- 2 % . With F(ab')2 fragments of 7E3 given as a bolus of 1.2 mg/kg , which saturated 70 % of the platelet P08514 /IIIa receptors and prolonged the bleeding to more than 30 min , lysis was not significantly increased over background . Combination of 0.3 or 0.6 mg/kg of 7E3-F(ab')2 with either 0.03 or 0.06 mg/kg of rt-PA did not produce more lysis than obtained with a comparable dose of rt-PA alone . No significant changes in plasma fibrinogen or alpha 2-antiplasmin were observed with either agent alone or with the combination . It is concluded that extensive inhibition of platelet aggregation does not potentiate the thrombolytic effect of rt-PA in this venous thrombosis model . Substance P promotes expansion of human mesenteric preadipocytes through proliferative and antiapoptotic pathways . White adipose tissue is intimately involved in the regulation of immunity and inflammation . We reported that human mesenteric preadipocytes express the DB05875 ( SP ) -mediated neurokinin-1 receptor ( P25103 ) , which signals proinflammatory responses . Here we tested the hypothesis that SP promotes proliferation and survival of human mesenteric preadipocytes and investigated responsible mechanism(s) . Preadipocytes were isolated from mesenteric fat biopsies during gastric bypass surgery . Proliferative and antiapoptotic responses were delineated in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium ( MTS ) , bromodeoxyuridine ( BrdU ) , caspase-3 , and TUNEL assays , as well as Western immunoanalysis . SP ( 10(-7) M ) increased MTS and proliferation ( BrdU ) and time dependently ( 15-30 min ) induced Akt , P01133 receptor , IGF receptor , integrin alphaVbeta3 , phosphatidylinositol 3-kinase , and PKC-theta phosphorylation . Furthermore , pharmacological antagonism of Akt and PKC-theta activation significantly attenuated SP-induced preadipocyte proliferation . Exposure of preadipocytes to the proapoptotic P48023 ( P48023 , 100 microM ) resulted in nuclear DNA fragmentation ( TUNEL assay ) , as well as increased cleaved poly ( ADP-ribose ) polymerase , cleaved caspase-7 , and caspase-3 expression . Cotreatment with SP almost completely abolished these responses in a P25103 -dependent fashion . SP ( 10(-7) M ) also time dependently stimulated expression 4E binding protein 1 and phosphorylation of P08133 S6 kinase , which increased protein translation efficiency . SP increases preadipocyte viability , reduces apoptosis , and stimulates proliferation , possibly via cell cycle upregulation and increased protein translation efficiency . SP-induced proliferative and antiapoptotic pathways in fat depots may contribute to development of the creeping fat and inflammation characteristic of Crohn 's disease . Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation . The platelet integrin glycoprotein ( GP ) IIb/IIIa , which mediates platelet aggregation , has been the target for novel antiplatelet agents , the P08514 /IIIa antagonists . Several P08514 /IIIa antagonists have been developed based on the peptide RGDS present in adhesion proteins , including the principle ligand fibrinogen . The apoptosis enzyme , procaspase-3 , contains an RGD-recognition sequence and is activated by RGDS . We examined the effects of RGDS and several P08514 /IIIa antagonists on cell death and procaspase-3 activation in rat neonatal cardiomyocytes . These antagonists do not recognize rat integrins , yet RGDS , orbofiban , and xemilofiban induced dose-dependent apoptosis and procaspase-3 activation in cardiomyocytes over 72 h , particularly under hypoxic conditions . Scrambled peptide , the monoclonal antibody 7E3 or integrelin ( a peptide containing a KGD sequence ) , had little or no effect . Immunoprecipitation of procaspase-3 followed by treatment with the compounds showed that procaspase-3 was activated directly by RGDS , orbofiban , xemilofiban , and by monoclonal DB00054 , the latter demonstrating that compounds must enter cells to induce apoptosis through caspase activation . DB00063 had no effect . Binding studies with (3)H-SC52012B , a P08514 /IIIa antagonist analogue of orbofiban , showed no specific binding to cardiomyocytes , but the radioligand accumulated intracellularly over 72 h . (3)H-SC52012B also bound directly to human recombinant caspase-3 ( K(d) , 59 +/- 2 nm ) , and this was prevented by orbofiban , xemilofiban , and the monoclonal DB00054 but not by integrelin . Finally confocal microscopy showed that RGDS co-localized with caspase-3 inside the cell . These data show that RGDS and its mimetics induce cardiomyocyte apoptosis by direct activation of procaspase-3 . Suppression of autoreactive T-cell response to glycoprotein IIb/IIIa by blockade of P25942 /CD154 interaction : implications for treatment of immune thrombocytopenic purpura . The potential immunosuppressive effect of an anti-CD154 monoclonal antibody ( mAb ) on the pathogenic autoreactive T-cell response was evaluated using an in vitro culture system with glycoprotein IIb/IIIa ( P08514 /IIIa ) -reactive T cells from patients with immune thrombocytopenic purpura ( ITP ) . The anti-CD154 mAb did not inhibit T-cell proliferation , but suppressed anti- P08514 /IIIa antibody production , in bulk peripheral blood mononuclear cell cultures stimulated with P08514 /IIIa . Repeated antigenic stimulation of P08514 /IIIa-reactive P01730 (+) T-cell lines in the presence of anti-CD154 mAb resulted in the loss of proliferative capacity and helper function for promoting anti- P08514 /IIIa antibody production . These anergic T-cell lines showed a cytokine profile of low interferon gamma and high interleukin 10 and suppressed anti- P08514 /IIIa antibody production . Our results indicate that blockade of the P25942 /CD154 interaction induces generation of autoantigen-specific anergic P01730 (+) T cells with regulatory function and could be a therapeutic option for suppressing pathogenic autoimmune responses in patients with ITP . A role for exposed mannosylations in presentation of human therapeutic self-proteins to P01730 + T lymphocytes . Several therapeutic self-proteins elicit immune responses when administered to patients . Such adverse immune responses reduce drug efficacy . To induce an immune response , a protein must interact with different immune cells , including antigen-presenting cells , T cells , and B cells . Each cell type recognizes distinct immunogenic patterns on antigens . Mannose-terminating glycans have been identified as pathogen-associated molecular patterns that are essential for internalization of microbes by antigen-presenting cells , leading to presentation . Here , we have investigated the importance of exposed mannosylation on an immunogenic therapeutic self-protein , procoagulant human factor VIII ( FVIII ) . Administration of therapeutic FVIII to hemophilia A patients induces inhibitory anti-FVIII antibodies in up to 30 % of the cases . We demonstrate that entry of FVIII into human dendritic cells ( DC ) leading to T cell activation , is mediated by mannose-terminating glycans on FVIII . Further , we identified macrophage mannose receptor ( CD206 ) as a candidate endocytic receptor for FVIII on DC . Saturation of mannose receptors on DC with mannan , and enzymatic removal of mannosylated glycans from FVIII lead to reduced T cell activation . The interaction between FVIII and CD206 was blocked by P04275 , suggesting that , under physiological conditions , the intrinsic mannose-dependent immunogenicity of FVIII is quenched by endogenous immunochaperones . These data provide a link between the mannosylation of therapeutic self-proteins and their iatrogenic immunogenicity . Such a link would be of special relevance in the context of replacement therapy where mechanisms of central and peripheral tolerance have not been established during ontogeny because of the absence of the antigen . A bispecific antifibrin-antiplatelet urokinase conjugate ( BAAUC ) induces enhanced clot lysis and inhibits platelet aggregation . Thrombolysis is well established in the treatment of acute myocardial infarction . However , clinical application of thrombolytic agents has limitations with respect to efficacy and specificity . To achieve highly effective and at the same time clot-selective plasminogen activation urokinase was coupled to a bispecific antibody consisting of the monovalent Fab ' from the antifibrin monoclonal antibody 59D8 and the monovalent Fab ' from the anti-glycoprotein P08514 /IIIa monoclonal antibody DB00054 . The bispecific antifibrin-antiplatelet urokinase conjugate ( BAAUC ) was synthesized and characterized . Assays with either immobilized platelets , P08514 /IIIa or fibrin showed an increase in plasminogen activation compared to uncoupled urokinase by 10-fold , 58-fold and 13-fold , respectivley ( p < 0.0001 each ) . In vitro clot lysis was performed on platelet-rich and fibrin-rich clots and revealed an up to 5-fold higher potency of BAAUC compared to uncoupled urokinase ( p < 0.0001 ) . In vitro platelet aggregation was effectively inhibited by the hybrid molecule , whereas urokinase had no effect . BAAUC and two monospecific urokinase-conjugates , UK-59D8-IgG and UK-7E3-(Fab')2 were compared with each other with regard to similar tests . In vitro clot assays with platelet-rich and platelet-poor clots were performed . BAAUC achieved a significantly higher plasminogen activation compared to each of the monospecific conjugates ( p < 0.05 , respectively ) . We conclude that BAAUC , a bispecific plasminogen activator with antifibrin and antiplatelet properties has the potency to lyse both fibrin-rich and platelet-rich thrombi with high efficacy and to effectively inhibit platelet aggregation . Role of Q14116 in overt pain-like behaviour in mice . There are evidences that targeting Q14116 might be beneficial to inhibit inflammatory symptoms , including hypernociception ( decrease in nociceptive threshold ) . The mechanism of Q14116 mechanical hypernociception depends on endothelin in rats and mice . However , the role of Q14116 in overt pain-like behaviour remains undetermined . Therefore , we addressed the role of Q14116 in writhing response induced by intraperitoneal ( i.p. ) injection of phenyl-p-benzoquinone ( PBQ ) and acetic acid in mice . Firstly , it was detected that PBQ and acetic acid i.p. injection induced a dose-dependent number of writhes in Balb/c mice . Subsequently , it was observed that the PBQ - but not the acetic acid-induced writhes were diminished in Q14116 deficient ( ( -/- ) ) mice . Therefore , considering that P01579 , endothelin and prostanoids mediate Q14116 -induced mechanical hypernociception , we also investigated the role of these mediators in the same model of writhing response in which Q14116 participates . It was noticed that PBQ-induced writhes were diminished in P01579 (-/-) mice and by the treatment with DB00559 ( mixed endothelin P25101 /ETB receptor antagonist ) , BQ 123 ( cyclo[DTrp-DAsp-Pro-DVal- DB00149 ] , selective endothelin P25101 receptor antagonist ) , BQ 788 ( N-cys-2,6 dimethylpiperidinocarbonyl-l-methylleucyl-d-1-methoxycarboyl-d-norleucine , selective endothelin ETB receptor antagonist ) or indomethacin ( cycloxigenase inhibitor ) . Thus , Q14116 , P01579 , endothelin acting on endothelin P25101 and ETB receptors , and prostanoids mediate PBQ-induced writhing response in mice . To conclude , these results further advance the understanding of the physiopathology of overt pain-like behaviour , and suggest for the first time a role for Q14116 in writhing response in mice . Genetic polymorphisms for the study of multifactorial stroke . Single-gene disorders explain only a minority of stroke cases . Stroke represents a complex trait , which is usually assumed to be polygenic . On this topic , the role of a wide number of candidate genes has been investigated in stroke through association studies , with controversial results . Therefore , it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke . This review is an update and an extensive analysis of the more recent association studies conducted in stroke . We evaluated a number of studies on several candidate genes ( including P12259 , F2 , P02671 / P02675 / P02679 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / Q9UKI9 / P04054 / P08514 , P17301 , P07359 , P12821 , AGT , NOS3 , P02649 , P06858 , P27169 , Q08499 , P20292 , P42898 , Q99707 , and P35520 ) , providing a final panel of genes and molecular variants . We categorized this panel in relation to the degree of association with stroke , supported by the results of meta-analyses and case-control studies . Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses . Dynamics of P08514 /IIIa-mediated platelet-platelet interactions in platelet adhesion/thrombus formation on collagen in vitro as revealed by videomicroscopy . The conventional description of platelet interactions with collagen-coated surfaces in vitro , based on serial static measurements , is that platelets first adhere and spread to form a monolayer and then recruit additional layers of platelets . To obtain dynamic information , we studied gravity-driven platelet deposition in vitro on purified type 1 collagen by video phase-contrast microscopy at 22 degrees C . With untreated human and wild-type mouse platelets , soon after the initial adhesion of a small number of " vanguard " platelets , " follower " platelets attached to the spread-out vanguard platelets . Follower platelets then adhered to and spread onto nearby collagen or over the vanguard platelets . Thus , thrombi formed as a concerted process rather than as sequential processes . Treatment of human platelets with monoclonal antibody ( mAb ) DB00054 ( anti- P08514 /IIIa ( alphaIIbbeta3 ) + alphaVbeta3 ) or tirofiban ( anti- P08514 /IIIa ) did not prevent platelet adhesion but nearly eliminated the deposition of follower platelets onto vanguard platelets and platelet thrombi . Similar results were obtained with Glanzmann thrombasthenia platelets . Wild-type mouse platelets in the presence of mAb 1B5 ( anti- P08514 /IIIa ) and platelets from beta3-null mice behaved like human platelets in the presence of 7E3 or tirofiban . Deposition patterns of untreated human and wild-type mouse platelets were consistent with random distributions under a Poisson model , but those obtained with 7E3- and tirofiban-treated human platelets , 1B5-treated mouse platelets , or beta3-null platelets demonstrated a more uniform deposition than predicted . Thus , in this model system , absence or blockade of P08514 /IIIa receptors interferes with thrombus formation and alters the pattern of platelet deposition . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . The high molecular mass , glycoprotein Ib-binding protein flavocetin-A induces only small platelet aggregates in vitro . The direct effects of snake venom glycoprotein ( GP ) Ib-binding proteins on platelet receptors during the formation of platelet aggregates were determined by a particle counting method using light scattering . Flavocetin-A induces small platelet aggregates , but not medium or large ones . However , neither jararaca GPIb-BP nor tokaracetin induce platelet aggregation . The flavocetin-A dose-response curve for formation of small aggregates is bell-shaped , with maximal effect at 1 to 2 microg/mL . The formation of small aggregates was not observed when fixed human platelets were used . Jararaca GPIb-BP , the anti-GPIb monoclonal antibody GUR83-35 , prostaglandin I2 , and ethylene diamine-N,N-dimethylformamide all inhibited flavocetin-A-induced small aggregate formation , but acetylsalicylic acid did not . Furthermore , anti- P08514 /IIIa monoclonal antibodies , DB00054 , and YM337 significantly but partially inhibited aggregate formation , but the anti- P04275 monoclonal antibody NMC-4 had no effect . The formation of small aggregates required extracellular calcium , but flavocetin-A did not elevate cytosolic calcium . These results suggest that flavocetin-A binds to intact platelets , initiating platelet responses and inducing platelet aggregate formation by cross-linking platelets . Consequently , flavocetin-A may be a useful tool to study the mechanism of GPIb-mediated platelet activation and the structure-function relationships of GPIb . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Repetitive profound thrombocytopenia after treatment with tirofiban : a case report . The P08514 /IIIa inhibitors are used in the acute coronary syndromes and interventional cardiology as antiplatelet agents . These drugs induce thrombocytopenia in approximately 1-5 % of patients . Thrombocytopenia is rapid in onset and antibody mediated . DB00054 is associated with higher incidence of thrombocytopenia than eptifibatide and tirofiban . Profound thrombocytopenia has reportedly been an issue with abciximab , but not with tirofiban . We reported a case of acute profound thrombocytopenia due to on tirofiban treatment in the same patient at two different times . Factor XIIIa binding to activated platelets is mediated through activation of glycoprotein IIb-IIIa . Stabilization of a clot is dependent on fibrin cross-linking mediated by the transglutaminase , factor XIIIa ( FXIIIa ) . In addition to fibrin stabilization , FXIIIa acts on a number of platelet-reactive proteins , including fibronectin and vitronectin , as well as the platelet proteins , glycoprotein ( GP ) IIb-IIIa , myosin , and actin . However , conditions inducing the platelet-activation dependent binding of FXIIIa have not been characterized nor have the sites mediating FXIIIa binding been identified . The generation of FXIIIa and consequent detection of FXIIIa on the platelet surface were compared with other thrombin-induced activation events ; the rate at which FXIIIa bound to activated platelets was much slower than platelet degranulation or fibrin(ogen) binding . Whereas platelets could be rapidly induced to express a functional receptor for FXIIIa , the rate of FXIIIa binding to platelets is limited by the rate of conversion of FXIII to FXIIIa . Immunoprecipitation of radiolabeled platelets using polyclonal anti-FXIII A-chain antibody identified two proteins corresponding to P08514 and P05106 . Preincubation of intact platelets with DB00054 , a monoclonal antibody that blocks the fibrinogen binding site , or GRGDSP peptide inhibited FXIIIa binding by about 95 % when measured by flow cytometry ; FXIIIa binding to purified P08514 -IIIa was also inhibited by DB00054 . The binding of FXIIIa to purified P08514 -IIIa was enhanced by the addition of fibrinogen , but not by that of fibronectin or thrombospondin , suggesting that FXIIIa also binds to fibrinogen associated with the complex . These observations suggest that activated platelets bearing FXIIIa may enhance stabilization of platelet-rich thrombi through surface-localized cross-linking events . Comparative studies of a humanized anti-glycoprotein IIb/IIIa monoclonal antibody , YM337 , and abciximab on in vitro antiplatelet effect and binding properties . The effects of YM337 , the Fab fragment of a humanized anti-glycoprotein IIb/IIIa ( P08514 /IIIa ) monoclonal antibody C4G1 , on in vitro platelet function and binding properties were compared with those of abciximab , the Fab fragment of the human/murine chimeric anti- P08514 /IIIa monoclonal antibody DB00054 . Both agents completely inhibited platelet aggregation caused by all agonists tested except ristocetin . Further , both inhibited human platelet adhesion to P04275 , fibrinogen , fibronectin and subendothelial matrix with similar potency . DB09222 binding to washed platelets was dose-dependently inhibited by both agents . In binding assay using 125I-YM337 and 125I-abciximab , Kd values determined with platelet-rich plasma were 6.74 +/- 0.56 nM for YM337 and 6.65 +/- 1.45 nM for abciximab , and the number of binding sites were 42,700 +/- 3,000 for YM337 and 76,000 +/- 5,400 for abciximab . P08514 /IIIa was precipitated from the solubilized fraction of platelets by both agents . In contrast , integrin alphavbeta3 was precipitated from the solubilized fraction of human umbilical vein endothelial cells by abciximab but not by YM337 . DB09222 binding to purified P08514 /IIIa was dose-dependently inhibited by both agents . In contrast , vitronectin binding to purified integrin alphavbeta3 was dose-dependently inhibited by abciximab but not by YM337 , supporting the idea that abciximab reacts to integrin alphavbeta3 . Therefore , YM337 was suggested to bind to a different epitope of P08514 /IIIa from abciximab . These results suggest that YM337 specifically acts on platelet P08514 /IIIa receptors and has similar inhibitory properties on platelet aggregation and platelet adhesion to abciximab . Flow cytometric analysis of platelet activation throughout normal gestation . OBJECTIVE : To measure platelet activation in normal pregnancy , before and after stimulation with agonists , with a whole blood flow cytometric technique . METHODS : In a cross-sectional study , 5 mL of whole blood was collected from healthy volunteers ( nine in the first trimester , ten in the second trimester , 35 in the third trimester , and 32 nonpregnant controls ) . Platelets were treated with an agonist ( thrombin or U-46619 , a thromboxane A2 analogue ) or buffer and were exposed to saturating concentrations of monoclonal antibodies directed against platelet membrane glycoproteins ( GPs ) : DB00054 ( fibrinogen receptor P08514 /IIIa ) , P28222 ( alpha granule marker P16109 ) , and 6D1 ( P04275 receptor GPIb ) . Mean fluorescence intensity was determined for 5000 platelets per sample by using a flow cytometer . RESULTS : In the absence of agonist , no significant difference between groups was found in antibody binding . At no stage of pregnancy were circulating activated platelets detected . Platelets from third-trimester subjects bound significantly less 7E3 than platelets of controls or of first- or second-trimester subjects after stimulation with high-dose thrombin ( P < .05 for all comparisons ) . Down-regulation of 6D1 on platelets after stimulation with high-dose U-46619 was significantly greater in third-trimester gravidas than in controls or first-trimester subjects ( P < .05 ) . CONCLUSION : Pregnancy does not increase the percentage of activated platelets in the circulation . Platelet reactivity is altered in the third trimester , as evidenced by decreased antibody binding to fibrinogen receptor epitope and enhanced down-regulation of a P04275 receptor epitope . Conjugation and evaluation of 7E3 x P4B6 , a chemically cross-linked bispecific F(ab')2 antibody which inhibits platelet aggregation and localizes tissue plasminogen activator to the platelet surface . A bispecific F(ab')2 monoclonal antibody which recognizes both the platelet P08514 /IIIa receptor and human tissue plasminogen activator was produced to target tPA to platelets for enhancement of thrombolysis . A stable , thioether-cross-linked bispecific F(ab')2 ( 7E3 X P4B6 ) combining the P08514 /IIIa-specific monoclonal antibody DB00054 , which inhibits platelet aggregation , and a nonneutralizing anti-tPA monoclonal antibody ( P4B6 ) was produced . This was performed by coupling each of the parental Fab ' moieties with the homobifunctional cross-linker bis ( maleimido methyl ) ether ( BMME ) . 7E3 X P4B6 was sequentially purified using gel-filtration chromatography and hydrophobic interaction ( HIC ) HPLC . HIC was shown to completely resolve each of the parental F(ab')2 species from the bispecific one . 7E3 X P4B6 was shown to retain completely each of the parental immunoreactivities in P08514 /IIIa and tPA binding EIA 's . The bispecific antibody inhibited platelet aggregation in vitro at levels comparable to those for 7E3 Fab . Recruitment of tPA activity to washed human platelets was demonstrated using the S-2251 chromogenic substrate assay . 7E3 X P4B6 recruited 12-fold more tPA to the washed platelets than a mixture of the parental F(ab')2 molecules used as controls . c7E3 Fab inhibits human tumor angiogenesis in a SCID mouse human skin xenograft model . The alphavbeta3 integrin plays an important role in tumor growth and angiogenesis . Inhibition of this receptor by intact bivalent antibodies has been shown to inhibit angiogenesis and tumor growth . In this study we tested the chimeric Fab of DB00054 ( c7E3 Fab ) , an antibody reactive with human platelet P08514 /IIIa and alphavbeta3 to determine if it would inhibit in vivo angiogenesis and tumor growth in a SCID mouse/human skin tumor growth and angiogenesis model . c7E3 Fab inhibited human tumor angiogenesis and tumor growth . These data suggest monovalent antibody fragments devoid of antibody effector function can have efficacy in preclinical models of angiogenesis . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . A chimeric murine/human antibody Fab fragment directed against the platelet P08514 /IIIa receptor enhances and sustains arterial thrombolysis with recombinant tissue-type plasminogen activator in baboons . Inhibition of the platelet glycoprotein ( GP ) IIb/IIIa receptor with the murine monoclonal antibody 7E3 abolishes ex vivo platelet aggregation , reduces thrombogenicity , and sustains arterial recanalization with recombinant tissue-type plasminogen activator ( rt-PA ) . A chimeric murine/human Fab fragment of DB00054 ( c7E3-Fab ) has a markedly reduced immunogenicity , but its potency as an adjunct for thrombolysis with rt-PA has not been evaluated . The effects of a single intravenous bolus injection of aspirin ( 17 mg/kg ) or c7E3-Fab ( 0.45 mg/kg ) on thrombolysis and reocclusion induced with rt-PA were studied in groups of six baboons with femoral arterial thrombosis and superimposed high-grade stenosis . This dose of c7E3-Fab blocked 96 +/- 1 % of the platelet P08514 /IIIa receptors and abolished ADP-induced platelet aggregation . Bolus intravenous injections of rt-PA ( 0.25 mg/kg ) were repeated at 15-minute intervals until reperfusion occurred ( maximum of four injections ) . In the aspirin group , reperfusion was obtained within 51 +/- 16 minutes ( mean +/- SD ) but was rapidly followed by reocclusion within 6 +/- 9 minutes and by cyclic reflow and reocclusion . In the c7E3-Fab group , reperfusion was obtained within 25 +/- 8 minutes ( P < .01 versus aspirin group ) and was associated with a delayed reocclusion of 63 +/- 63 minutes ( P < .05 versus aspirin group ) . Template bleeding times remained unchanged in the aspirin/rt-PA group but were markedly prolonged ( to > 30 minutes ) in the c7E3-Fab/rt-PA group. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00054 , eptifibatide , and tirofiban exhibit dose-dependent potencies to dissolve platelet aggregates . Platelet P08514 /IIIa antagonists are not only used to prevent platelet aggregation , but also in combination with thrombolytic agents for the treatment of coronary thrombi . Recent data indicate a potential of abciximab alone to dissolve thrombi in vivo . We investigated the potential of abciximab , eptifibatide , and tirofiban to dissolve platelet aggregates in vitro . DB00640 diphosphate ( ADP ) -induced platelet aggregation could be reversed in a concentration-dependent manner by all three P08514 /IIIa antagonists when added after the aggregation curve reached half-maximal aggregation . The concentrations chosen are comparable with in vivo plasma concentrations in clinical applications . Disaggregation reached a maximum degree of 72.4 % using 0.5 microg/ml tirofiban , 91.5 % using 3.75 microg/ml eptifibatide , and 48.4 % using 50 microg/ml abciximab ( P < 0.05 , respectively ) . A potential fibrinolytic activity of the P08514 /IIIa antagonists was ruled out by preincubation with aprotinin or by a plasma clot assay . A stable model Chinese hamster ovary ( CHO ) cell line expressing the activated form of P08514 /IIIa was used to confirm the disaggregation capacity of P08514 /IIIa antagonists found in platelets . Not only abciximab , but also eptifibatide and tirofiban have the potential to disaggregate newly formed platelet clusters in vitro . Because enzyme-dependent fibrinolysis does not appear to be involved , competitive removal of fibrinogen by the receptor antagonists is the most likely mechanism . Prevention of rethrombosis after coronary thrombolysis in a chronic canine model . I . Adjunctive therapy with monoclonal antibody 7E3 F(ab')2 fragment . We examined the efficacy of the monoclonal antibody ( MoAb ) 7E3 F(ab')2 fragment , an inhibitor of the platelet glycoprotein (GP)IIb/IIIa receptor , to prevent coronary artery rethrombosis after successful thrombolysis with rt-PA . The circumflex coronary artery of anesthetized dogs was instrumented with a flow probe , an electrode , and a stenosis . After recovery from the surgical procedure , the animals were reanesthetized on post-operative day 9 , and vessel wall injury was induced with current applied to the intimal surface of the circumflex coronary artery . The resulting occlusive thrombus was aged for 30 min , and recombinant tissue plasminogen activator ( rt-PA ) was administered . The animals were allocated to receive either placebo or a single dose of DB00054 [ 0.8 mg/kg intravenous ( i.v. ) bolus ] as the sole adjunctive agent . Ex vivo platelet function and coronary artery blood flow velocity were recorded on each of 5 consecutive days . Reocclusion and mortality were reduced significantly in animals treated with 7E3 as compared with the placebo-treated group . Significant inhibition of ex vivo platelet aggregation persisted for 48 h after a single injection of DB00054 . The MoAb 7E3 F(ab')2 fragment is effective as the sole adjunctive agent with rt-PA for prevention of rethrombosis . The present study is unique in that it examined the efficacy of P08514 /IIIa inhibition in an experimental model for an extended time , demonstrating the duration of antiplatelet therapy required to prevent rethrombosis after thrombolysis . P08514 /IIIa antagonists and other anti-integrins . Platelet aggregation involves the binding of adhesive proteins ( fibrinogen , P04275 ) to the alphaIIbbeta3 integrin , which assumes a high-affinity state for adhesive proteins during platelet activation . The occupied integrin sends signals back into the platelet , and the bound adhesive protein forms the bridges linking platelets together . Anti-integrin therapy is designed to inhibit this process in arterial thrombosis . DB00054 , mouse-human chimeric Fab fragments , blocks platelet aggregation and provides proven clinical benefit in acute situations such as in patients with unstable angina undergoing angioplasty or stenting . DB00063 and tirofiban are small molecular mass inhibitors also in current use . In contrast , oral inhibitors of alphaIIbbeta3 have proved disappointing , provoking increased mortality without assuring an adequate blockade of alphaIIbbeta3 . The problems of using anti-integrin therapy are discussed in this article as are ways of improving its efficacity . Final thoughts provide ideas for a new generation of inhibitors . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Therapeutic Implications of a Specific Murine Monoclonal Antibody ( DB00054 ) to the Platelet Receptor P08514 /IIIa . The murine monoclonal antibody 7E3 blocks the platelet glycoprotein IIb/IIIa receptor , and is a potent inhibitor of platelet function in both animals and man . Animal models of the acute coronary syndromes suggest that 7E3 abolishes the in vivo formation of platelet thrombi , accelerates thrombolysis with tissue plasminogen activator , and prevents subsequent reocclusion . Human studies with 7E3 suggest that complete inhibition of platelet function may be safely undertaken for periods of up to 36 h , and preliminary studies indicate effectiveness in the therapy of clinical unstable angina . Potential problems with its use include immunogenicity and thrombocytopenia . The outcome of the acute coronary thrombotic syndromes , which include unstable angina , acute myocardial infarction and abrupt closure following coronary angioplasty , may be significantly improved with 7E3 therapy . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . New anticoagulant strategies . The limitations of standard heparin have prompted the development of a variety of newer antithrombotic agents . In fact , a LMWH preparation has recently been approved for clinical use in North America . Of these novel preparations , LMWH , the direct thrombin inhibitors , and inhibitors of P08514 -IIIa have been used clinically and are in advanced stages of evaluation . Not only is LMWH effective in the prevention of venous thromboembolic disease in high-risk patients , but its more predictable dose response makes it an ideal candidate for the treatment of venous thrombosis . Further studies are needed to determine whether LMWH is superior to standard heparin as adjunctive therapy in patients undergoing coronary thrombolysis or angioplasty . Particularly promising in the setting of arterial thrombosis are hirudin , hirulog , and DB00054 . With the encouraging results reported to date , it is likely that these agents will soon find their way into the treatment armamentarium of arterial thrombosis . Platelet-associated anti- P08514 -IIIa autoantibodies in chronic immune thrombocytopenic purpura recognizing epitopes close to the ligand-binding site of glycoprotein ( GP ) IIb . Localization of epitopes for platelet-associated ( PA ) anti- P08514 -IIIa ( alpha(IIb)beta(3) ) autoantibodies in chronic immune thrombocytopenic purpura remains elusive . Previous studies suggest that PA antibodies recognize the tertiary structure of intact glycoprotein ( GP ) IIb-IIIa . To localize their epitopes using antigen-capture enzyme-linked immunosorbent assay ( ELISA ) , the reactivity of 34 PA anti- P08514 -IIIa antibodies was examined with recombinant P08514 -IIIa having a defect in ligand-binding sites in either P08514 or P05106 , and no major conformational change was induced : KO variant P08514 -IIIa was attributed to a 2-amino acid insertion between residues 160 and 161 in the W3 4-1 loop in P08514 , and P62158 variant P08514 -IIIa was attributed to D119Y in P05106 . In one third ( 11 of 34 ) of the patients , PA antibodies showed a marked decrease ( less than 50 % ) in reactivity with KO compared with wild-type P08514 -IIIa . Their reactivity was also impaired against GPIIbD163A-IIIa . In sharp contrast , they reacted normally with P62158 P08514 -IIIa . OP-G2 , a ligand-mimetic monoclonal antibody , markedly inhibited their binding to P08514 -IIIa in patients with impaired binding to KO P08514 -IIIa , but small P08514 -IIIa antagonists did not . In addition , a newly developed sensitive ELISA indicated that autoantibodies showing impaired binding to KO are more potent inhibitors for fibrinogen binding . The present data suggest that certain PA anti- P08514 -IIIa autoantibodies recognize epitopes close to the ligand-binding site in P08514 , but not in P05106 . Monoclonal antibody against the platelet glycoprotein ( GP ) IIb/IIIa receptor prevents coronary artery reocclusion after reperfusion with recombinant tissue-type plasminogen activator in dogs . Localized thrombosis was produced in the left anterior descending ( LAD ) coronary artery of open chest dogs by constricting a segment so as to produce greater than 90 % stenosis ( reducing blood flow to 40 +/- 10 % of baseline ) , and placing a thrombus in the segment immediately proximal to the stenosis by inducing endothelial cell injury and instilling a mixture of blood and thrombin . Intravenous infusion of recombinant tissue-type plasminogen activator ( rt-PA ) at a rate of 15-30 micrograms/kg per min for 30 or 60 min in eight dogs induced coronary artery reperfusion within 23 +/- 7 min ( mean +/- SD ) , but reocclusion occurred despite heparin anticoagulation in all but one of these dogs within 7 +/- 5 min . Intravenous injection of 0.8 mg/kg of the F(ab')2 fragment of a monoclonal antibody ( DB00054 ) directed against the platelet P08514 /IIIa receptor , prevented reocclusion in 10/10 dogs during an observation period of 2 h ( P less than 0.001 vs. rt-PA alone ) . The antibody abolished ADP-induced platelet aggregation and markedly prolonged the bleeding time . Intravenous aspirin or dipyridamole prevented reocclusion for 1 h or more in only 2/7 and 1/6 dogs , respectively . We conclude that the monoclonal antibody is very effective in preventing reocclusion after successful thrombolysis of occluded coronary arteries with rt-PA . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) G-protein-gated inwardly rectifying potassium channels regulate ADP-induced P47712 activity in platelets through Src family kinases . ADP-induced TXA2 generation requires the costimulation of P47900 , Q9H244 , and the P08514 /IIIa receptors . Signaling events downstream of the P2Y receptors that contribute to ADP-induced TXA2 generation have not been clearly delineated . In this study , we have investigated the role of G-protein-gated inwardly rectifying potassium channels ( GIRKs ) , a recently identified functional effector for the Q9H244 receptor , in the regulation of ADP-induced TXA2 generation . At 10-microM concentrations , the 2 structurally distinct GIRK channel blockers , SCH23390 and U50488H , caused complete inhibition of ADP-induced P47712 phosphorylation and TXA2 generation , without affecting the conversion of AA to TXA2 or ADP-induced primary platelet aggregation in aspirin-treated platelets . In addition , Src family kinase selective inhibitors abolished 2MeSADP-mediated P47712 phosphorylation and TXA2 generation . Furthermore , these GIRK channel blockers completely blocked Gi-mediated Src kinase activation , suggesting that GIRK channels are upstream of Src family tyrosine kinase activation . In weaver mouse platelets , which have dysfunctional P48051 subunits , ADP-induced TXA2 generation was impaired . However , we did not observe any defect in 2MeSADP-induced platelet functional responses in P48051 -null mouse platelets , suggesting that functional channels composed of other GIRK subunits contribute to ADP-induced TXA2 generation , via the regulation of the Src and P47712 activity . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane .

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interacts_with DB00208?

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Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Nerve agent exposure elicits site-specific changes in protein phosphorylation in mouse brain . Organophosphorus ( OP ) compounds cause toxic symptoms , including convulsions , coma , and death , as the result of irreversible inhibition of acetylcholinesterase ( P22303 ) . The development of effective treatments to block these effects and attenuate long-term cognitive and motor disabilities that result from OP intoxication is hampered by a limited understanding of the CNS pathways responsible for these actions . We employed a candidate method ( called CNSProfile ) to identify changes in the phosphorylation state of key neuronal phosphoproteins evoked by the OP compound , diisopropyl fluorophosphate ( DB00677 ) . Focused microwave fixation was used to preserve the phosphorylation state of phosphoproteins in brains of DB00677 -treated mice ; hippocampus and striatum were analyzed by immunoblotting with a panel of phospho-specific antibodies . DB00677 exposure elicited comparable effects on phosphorylation of brain phosphoproteins in both C57BL/6 and FVB mice . DB00677 treatment significantly altered phosphorylation at regulatory residues on glutamate receptors , including Serine897 ( S897 ) of the Q9UHB4 DB01221 receptor . Q9UHB4 phosphorylation was bi-directionally regulated after DB00677 in striatum versus hippocampus . Q9UHB4 phosphorylation was reduced in striatum , but elevated in hippocampus , compared with controls . Q9UD71 phosphorylation in striatum was selectively increased at the Cdk5 kinase substrate , Threonine75 ( T75 ) . Phencynonate hydrochloride , a muscarinic cholinergic antagonist , prevented seizure-like behaviors and the observed changes in phosphorylation induced by DB00677 . The data reveal region-specific effects of nerve agent exposure on intracellular signaling pathways that correlate with seizure-like behavior and which are reversed by the muscarinic receptor blockade . This approach identifies specific targets for nerve agents , including substrates for Cdk5 kinase , which may be the basis for new anti-convulsant therapies . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Vascular transcriptional alterations produced by juvenile obesity in Ossabaw swine . We adopted a transcriptome-wide microarray analysis approach to determine the extent to which vascular gene expression is altered as a result of juvenile obesity and identify obesity-responsive mRNAs . We examined transcriptional profiles in the left anterior descending coronary artery ( LAD ) , perivascular fat adjacent to the LAD , and descending thoracic aorta between obese ( n = 5 ) and lean ( n = 6 ) juvenile Ossabaw pigs ( age = 22 wk ) . Obesity was experimentally induced by feeding the animals a high-fat/high-fructose corn syrup/high-cholesterol diet for 16 wk . We found that expression of 189 vascular cell genes in the LAD and expression of 165 genes in the thoracic aorta were altered with juvenile obesity ( false discovery rate ≤ 10 % ) with an overlap of only 28 genes between both arteries . Notably , a number of genes found to be markedly upregulated in the LAD of obese pigs are implicated in atherosclerosis , including P13686 , P61626 , O95715 , P02649 , Q13093 , P17931 , P10451 , P05107 , P04839 , and Q9H244 . Furthermore , pathway analysis revealed the induction of proinflammatory and pro-oxidant pathways with obesity primarily in the LAD . Gene expression in the LAD perivascular fat was minimally altered with juvenile obesity . Together , we provide new evidence that obesity produces artery-specific changes in pretranslational regulation with a clear upregulation of proatherogenic genes in the LAD . Our data may offer potential viable drug targets and mechanistic insights regarding the molecular precursors involved in the origins of overnutrition and obesity-associated vascular disease . In particular , our results suggest that the oxidized LDL/ P78380 /NF-κB signaling axis may be involved in the early initiation of a juvenile obesity-induced proatherogenic coronary artery phenotype . Gq-mediated Akt translocation to the membrane : a novel PIP3-independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol-3,4,5-trisphosphate ( PIP3 ) -dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5'-diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3-kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K/PIP3/Gi-independent mechanism . An Akt scaffolding protein , P38936 -activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi/PI3K/PIP3-independent mechanism . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Poly(ADP-ribose) polymerase-1 is a nuclear epigenetic regulator of mitochondrial DNA repair and transcription . Poly(ADP-ribose) polymerase-1 ( P09874 ) is a NAD-consuming enzyme with an emerging key role in epigenetic regulation of gene transcription . Although P09874 expression is characteristically restricted to the nucleus , a few studies report the mitochondrial localization of the enzyme and its ability to regulate organelle functioning . Here , we show that , despite exclusive nuclear localization of P09874 , mitochondrial homeostasis is compromised in cell lines exposed to P09874 pharmacological inhibitors or small interfering RNA . P09874 suppression reduces integrity of mitochondrial DNA ( mtDNA ) , as well as expression of mitochondria-encoded respiratory complex subunits P23219 , P35354 , and ND-2 . Accordingly , P09874 localizes at promoters of nuclear genes encoding both the mtDNA repair proteins P13051 , P12882 , and P27695 and the mtDNA transcription factors Q8WVM0 and Q9H5Q4 . It is noteworthy that poly(ADP-ribosyl)ation is required for nuclear gene expression of these mitochondrial proteins . Consistent with these findings , P09874 suppression impairs mitochondrial DB00171 production . Our results indicate that P09874 plays a central role in mitochondrial homeostasis by epigenetically regulating nuclear genes involved in mtDNA repair and transcription . These data might have important implications in pharmacology of P09874 inhibitors as well as clinical oncology and aging . Echicetin coated polystyrene beads : a novel tool to investigate GPIb-specific platelet activation and aggregation. P04275 /ristocetin ( P04275 /R ) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin , which also binds P04275 . These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets . Here , we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads , which specifically activate GPIb . We compared platelet activation induced by echicetin beads to P04275 /R . Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling . Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets , while under the same conditions P04275 /R treatment led only to αIIbβ3-independent platelet agglutination . The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation , while the total amount of echicetin used for activation is not critical . Echicetin beads induced strong phosphorylation of several proteins including p38 , P29323 and P31749 . Synergistic signaling via Q9H244 and thromboxane receptor through secreted ADP and TxA2 , respectively , were important for echicetin bead triggered platelet activation . Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation . Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways . A critical appraisal of the functional evolution of Q9H244 antagonists as antiplatelet drugs . Q9H244 receptor mediated inhibition of platelet aggregation is one of the most explored and exploited pathways in antiplatelet drug therapy to prevent ischemic events in patients undergoing percutaneous coronary intervention ( P05154 ) for the treatment of the acute coronary syndrome ( ACS ) . DB00208 , DB00758 , Prasugrel , DB08816 , DB06441 and Elinogrel are the Q9H244 inhibitors that act as antiplatelet drugs . In this review , the features of these drugs and the factors reported to be responsible for drug resistance or drug ineffectiveness were described . The features like drug metabolism , reversible or irreversible binding of drugs to their target protein and the mode of administration were observed to evolve along with the antiplatelet drugs . These features also include the drug-drug interactions , the pharmacogenetics and pharmacodynamics of Q9H244 inhibitors . We attempted to critically analyze how the desirable features were met by the Q9H244 inhibitors in the course of time . This review provides an overview of the evolution of Q9H244 inhibitors and may guide the researchers to develop better antiplatelet drugs in the future . Antiinflammatory and neurological activity of pyrithione and related sulfur-containing pyridine N-oxides from Persian shallot ( Allium stipitatum ) . ETHNOPHARMACOLOGICAL RELEVANCE : Persian shallot ( Allium stipitatum ) is a bulbous plant native to Turkey , Iran and Central Asia . It is frequently used in folk medicine for the treatment of a variety of disorders , including inflammation and stress . Antiinflammatory and neurological activities of pyrithione and four related sulfur-containing pyridine N-oxides which are prominent constituents of Allium stipitatum were tested . METHODS : The antiinflammatory activity was tested by the ability of the compounds to inhibit cyclooxygenase ( P23219 and P35354 ) , whereas the neurological activities were evaluated by assessing the compounds ability to inhibit monoamine oxidase-A ( P21397 ) and acetylcholinesterase ( P22303 ) . The compounds׳ affinity for the serotonin transport protein ( P31645 ) and the GABAA-benzodiazepine receptor were also investigated . RESULTS : 2-[(Methylthio)methyldithio]pyridine N-oxide showed very high antiinflammatory effects which are comparable with those of common pharmaceuticals ( IC₅₀ of 7.8 and 15.4 µM for P23219 and P35354 , respectively ) . On the other hand , neurological activities of the compounds were rather modest . Some compounds moderately inhibited P22303 ( IC₅₀ of 104-1041 µM ) and P21397 ( IC₅₀ of 98-241 µM ) and exhibited an affinity for the P31645 and GABAA-benzodiazepine receptor . CONCLUSIONS : Our findings may help to rationalize the wide use of Persian shallot for the treatment of inflammatory disorders . DB00171 stimulates O14788 expression through P47900 receptor-cyclo-oxygenase-dependent pathway in human periodontal ligament cells . BACKGROUND AND OBJECTIVE : Our previous study showed that human periodontal ligament cells responded to mechanical stress by increasing adenosine triphosphate ( DB00171 ) release , accompanied by the increased expression of O14788 and osteopontin . We found that the signaling pathway of mechanical stress-induced osteopontin was mediated through DB00171 /P2Y(1) receptor and Rho kinase activation but that of mechanical stress-induced O14788 was different . In this study , we further investigated the effect of extracellular DB00171 on the expression of O14788 and the mechanism involved . MATERIAL AND METHODS : Human periodontal ligament cells were treated with DB00171 ( 10-40 microm ) . The expressions of O14788 and cyclo-oxygenase 2 ( P35354 ) were examined by RT-PCR and western blot analysis . The level of prostaglandin E(2) was determined using ELISA . Signaling pathways were investigated by using inhibitors and antagonist . RESULTS : DB00171 induced the expression of O14788 . Indomethacin , an inhibitor of P36551 , could abolish the induction of O14788 expression , suggesting a P36551 -dependent mechanism . A DB02527 -dependent protein kinase inhibitor , H89 , and a nuclear factor kappaB ( NF kappaB ) inhibitor , pyrrolidine dithiocarbamate , inhibited O14788 expression , prostaglandin E(2) production and NF kappaB translocation . In addition , a specific P2Y(1) receptor antagonist , MRS2179 , and P2Y(1) small interfering RNA diminished the effect of DB00171 . CONCLUSION : Extracellular DB00171 stimulates O14788 expression in human periodontal ligament cells through a pathway dependent on the P2Y(1) receptor , DB02527 -dependent protein kinase , NF kappaB and P36551 . Our results suggest that , among the molecules responsible for the effect of mechanical stress , DB00171 participates in bone resorption or bone homeostasis by mediating its signal through the P2Y(1) receptor and the NF kappaB- P36551 - O14788 axis in periodontal tissue . Comparative effects of the anti-platelet drugs , clopidogrel , ticlopidine , and cilostazol on aspirin-induced gastric bleeding and damage in rats . AIMS : The present study compared the effects of frequently used anti-platelet drugs , such as clopidogrel , ticlopidine , and cilostazol , on the gastric bleeding and ulcerogenic responses induced by intraluminal perfusion with 25 mM aspirin acidified with 25 mM HCl ( acidified ASA ) in rats . MAIN METHODS : The stomach was perfused with acidified ASA at a rate of 0.4 ml/min for 60 min under urethane anesthesia , and gastric bleeding was measured as the concentration of hemoglobin in the luminal perfusate , which was collected every 15 min . DB00758 ( 10-100mg/kg ) , ticlopidine ( 10-300 mg/kg ) , or cilostazol ( 3-30 mg/kg ) was given p.o . 24h or 90 min before the perfusion of acidified ASA , respectively . KEY FINDINGS : Perfusion of the stomach with acidified ASA alone led to slight bleeding and lesions in the stomach . The pretreatment with clopidogrel , even though it did not cause bleeding or damage by itself , dose-dependently increased the gastric bleeding and ulcerogenic responses induced by acidified ASA . DB00208 also aggravated the severity of damage by increasing gastric bleeding , and the effects of ticlopidine at 300 mg/kg were equivalent to those of clopidogrel at 100mg/kg . In contrast , cilostazol dose-dependently decreased gastric bleeding and damage in response to acidified ASA . SIGNIFICANCE : These results demonstrated that clopidogrel and ticlopidine , Q9H244 receptor inhibitors , increased gastric bleeding and ulcerogenic responses to acidified ASA , to the same extent , while cilostazol , a phosphodiesterase III inhibitor , suppressed these responses . Therefore , cilostazol may be safely used in dual anti-platelet therapy combined with ASA , without increasing the risk of gastric bleeding . Cyclo-oxygenases and prostaglandins in acute inflammatory demyelination of the peripheral nerve . OBJECTIVE : To investigate the expression of cyclo-oxygenases ( P36551 ) , key enzymes in propagating inflammatory responses by converting arachidonic acid to prostaglandins , in inflammatory demyelinating disorders of the peripheral nervous system ( PNS ) . METHODS : Expression and distribution of P36551 messenger RNA ( mRNA ) and protein were studied in sural nerve biopsies , serum , and P04141 samples from patients with Guillain-Barré syndrome ( GBS ) , chronic inflammatory demyelinating polyradiculoneuropathy ( CIDP ) , or , for comparison , with vasculitic neuropathy ( VN ) , which is a inflammatory nondemyelinating disorder , and noninflammatory neuropathies ( Q8N4C6 ) using RT-PCR , immunohistochemistry , and immunoblotting . To confirm functional P35354 activity , the expression of prostaglandin E(2) ( PGE(2) ) and prostaglandin F(2alpha) ( P49763 (2alpha) ) was evaluated by ELISA ex vivo and in vitro . RESULTS : Whereas P23219 expression was unaltered in all investigated groups , a significant upregulation of P35354 mRNA was detected in sural nerves from patients with GBS , CIDP , or VN but not in control subjects with noninflammatory disorders . Macrophages were identified as its primary cellular source . Increased P35354 protein levels were detectable in serum and P04141 from all patients with GBS and , in smaller numbers only , in samples from patients with CIDP or VN but not from the Q8N4C6 group studied . Moreover , increased levels of PGE(2) and P49763 (2alpha) were measurable in sera from patients with GBS , CIDP , or VN and in cell culture supernatants from in vitro stimulated macrophages , indicative of P35354 activity . CONCLUSIONS : Cyclo-oxygenase-2 , expressed by macrophages , may generate prostaglandins during acute inflammatory demyelination of the peripheral nerve . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . ADP receptors -- targets for developing antithrombotic agents . Platelet P2 receptors -- P47900 , Q9H244 , and P51575 -- constitute the means by which adenine nucleotides can activate platelets . Coactivation of the Galphaq-coupled P47900 and Galphai2-coupled Q9H244 receptors is necessary for ADP-mediated platelet activation , which forms the basis of using P2 antagonists as antithrombotic drugs . P47900 receptor antagonists inhibit platelet activation , while P47900 knockout mice show longer bleeding times than normal mice but few other problems ; however , its ubiquitous expression in other tissues renders P47900 questionable as an antithrombotic target . The Q9H244 receptor is expressed nearly exclusively in platelets and brain , making it an attractive antithrombotic target . Antagonists for the Q9H244 receptor have been developed that either require metabolic activation to covalently inhibit Q9H244 and are irreversible , or simply are competitive in nature and thus reversible . DB00208 and clopidogrel are irreversible Q9H244 antagonists and have been repeatedly proven as clinical antithrombotic agents . In addition , a recently reported Q9H244 antagonist , CS-747 , shows promise as a future antithrombotic drug . The AR-C series of compounds represent reversible Q9H244 antagonists and have been used extensively to characterize the function of Q9H244 in platelets . Clinical studies show that AR-C69931MX is as effective as clopidogrel ; furthermore , the combination of AR-C69931MX ( cangrelor ) and clopidogrel confers greater antagonism of Q9H244 than either antagonist alone . The P51575 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change , and its inhibition or loss has little if any effect on hemostasis . A combination of P47900 and Q9H244 antagonists may represent an additional course of antithrombotic treatment . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . Nucleotide coronary vasodilation in guinea pig hearts . The role of P1 receptors and P47900 receptors in coronary vasodilator responses to adenine nucleotides was examined in the isolated guinea pig heart . Bolus arterial injections of nucleotides were made in hearts perfused at constant pressure . Peak increase in flow was measured before and after addition of purinoceptor antagonists . Both the P1 receptor antagonist 8-(p-sulfophenyl)theophylline and adenosine deaminase inhibited adenosine vasodilation . AMP-induced vasodilation was inhibited by P1 receptor blockade but not by adenosine deaminase or by the selective P47900 antagonist N6-methyl-2'-deoxyadenosine 3',5'-bisphosphate ( P59665 2179 ) . ADP-induced vasodilation was moderately inhibited by P1 receptor blockade and greatly inhibited by combined P1 and P47900 blockade . DB00171 -induced vasodilation was antagonized by P1 blockade but not by adenosine deaminase . Addition of P47900 blockade to P1 blockade shifted the DB00171 dose-response curve further rightward . It is concluded that in this preparation DB00171 -induced vasodilation results primarily from AMP stimulation of P1 receptors , with a smaller component from DB00171 or ADP acting on P47900 receptors . ADP-induced vasodilation is largely due to P47900 receptors , with a smaller contribution by AMP or adenosine acting via P1 receptors . AMP responses are mediated solely by P1 receptors . DB00640 contributes very little to vasodilation resulting from bolus intracoronary injections of DB00171 , ADP , or AMP . Contribution of the Q9H244 receptor-mediated pathway to platelet hyperreactivity in hypercholesterolemia . BACKGROUND : In hypercholesterolemia , platelets demonstrate increased reactivity and promote the development of cardiovascular disease . OBJECTIVE : This study was carried out to investigate the contribution of the ADP receptor Q9H244 -mediated pathway to platelet hyperreactivity due to hypercholesterolemia . METHODS : P01130 -deficient mice and C57Bl/6 wild-type mice were fed on normal chow and high-fat ( Western or Paigen ) diets for 8 weeks to generate differently elevated cholesterol levels . Q9H244 receptor-induced functional responses via G(i) signaling were studied ex vivo when washed murine platelets were activated by 2MeSADP and PAR4 agonist AYPGKF in the presence and absence of indomethacin . Platelet aggregation and secretion , α(IIb)β(3) receptor activation and the phosphorylation of extracellular signal-regulated protein kinase ( P29323 ) and Akt were analyzed . RESULTS : Plasma cholesterol levels ranged from 69 ± 10 to 1011 ± 185 mg dL(-1) depending on diet in mice with different genotypes . Agonist-dependent aggregation , dense and α-granule secretion and JON/A binding were gradually and significantly ( P < 0.05 ) augmented at low agonist concentration in correlation with the increasing plasma cholesterol levels , even if elevated thromboxane generation was blocked . These functional responses were induced via increased levels of G(i) -mediated P29323 and Akt phosphorylation in hypercholesterolemic mice vs. normocholesterolemic animals . In addition , blocking of the Q9H244 receptor by AR-C69931MX ( DB06441 ) resulted in strongly reduced platelet aggregation in mice with elevated cholesterol levels compared with normocholesterolemic controls . CONCLUSIONS : These data revealed that the Q9H244 receptor pathway was substantially involved in platelet hyperreactivity associated with mild and severe hypercholesterolemia . [ Thienopyridines in the treatment and prevention of cardiovascular diseases. Part I. DB00208 ] . In a series of articles the authors consider clinical pharmacology and experience of clinical application of blockers of platelet Q9H244 receptors , most well known representatives of which ticlopidine and clopidogrel according to chemical structure belong to thienopyridine derivatives . In the first communication pharmacodynamics and pharmacokinetics of the first thienopyridine ticlopidine are described in detail . Results of randomized studies in which cerebro and cardioprotective efficacy and safety of ticlopidine was studied in patients with cerebrovascular , peripheral artery diseases , and acute coronary syndromes are discussed . It has been established that ticlopidine is more effective and safe in patients having undergone coronary and femoral bypass surgery . Results of meta analyses have shown which evidence that ticlopidine is not less and may be more effective than clopidogrel in patients after coronary bypass surgery . Most frequent and most severe side effects of ticlopidine and measures of their prevention are also considered . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . Activation of Gi-coupled receptors releases a tonic state of inhibited platelet aggregation . Single-receptor pharmacology does not satisfactorily explain the physiology of the ADP-induced platelet aggregation response . It has been shown that , in addition to Gq-coupled receptor activation , one Gi-coupled receptor , either the ADP P2T or the alpha2-adrenoceptor , is required for elicitation of aggregation . The underlying mechanism of this action , however , has not been elucidated . By systematically assaying the entire time course of the aggregation and its fade using two methods of aggregometry , we have investigated the role of graded activation of these two Gi-coupled receptors . We demonstrate that constant activation of either of two Gq-coupled receptors , the ADP P47900 or the 5- Q13049 , and incremental activation of either of the two Gi-coupled receptors , tightly regulates the aggregation response in vitro , through the apparent release of a tonic inhibition of platelet aggregation . This tightly regulated release of inhibition , which appears analogous to the phenomena of disinhibition observed in the central nervous system , may be instrumental for the continuous adaptation of the aggregation response to variable physiological conditions . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Alteration of synaptic activity-regulating genes underlying functional improvement by long-term exposure to an enriched environment in the adult brain . BACKGROUND : Housing animals in an enriched environment ( EE ) enhances behavioral function . However , the mechanism underlying this EE-mediated functional improvement and the resultant changes in gene expression have yet to be elucidated . OBJECTIVES : We attempted to investigate the underlying mechanisms associated with long-term exposure to an EE by evaluating gene expression patterns . METHODS : We housed 6-week-old CD-1 ( ICR ) mice in standard cages or an EE comprising a running wheel , novel objects , and social interaction for 2 months . Motor and cognitive performances were evaluated using the rotarod test and passive avoidance test , and gene expression profile was investigated in the cerebral hemispheres using microarray and gene set enrichment analysis ( GSEA ) . RESULTS : In behavioral assessment , an EE significantly enhanced rotarod performance and short-term working memory . Microarray analysis revealed that genes associated with neuronal activity were significantly altered by an EE . GSEA showed that genes involved in synaptic transmission and postsynaptic signal transduction were globally upregulated , whereas those associated with reuptake by presynaptic neurotransmitter transporters were downregulated . In particular , both microarray and GSEA demonstrated that EE exposure increased opioid signaling , acetylcholine release cycle , and postsynaptic neurotransmitter receptors but decreased Na+ / Cl- -dependent neurotransmitter transporters , including dopamine transporter Slc6a3 in the brain . Western blotting confirmed that Q01959 , Q9UD71 ( Q9UD71 ) , and Q9H244 were largely altered in a region-specific manner . CONCLUSION : An EE enhanced motor and cognitive function through the alteration of synaptic activity-regulating genes , improving the efficient use of neurotransmitters and synaptic plasticity by the upregulation of genes associated with postsynaptic receptor activity and downregulation of presynaptic reuptake by neurotransmitter transporters . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . The thienopyridine derivatives ( platelet adenosine diphosphate receptor antagonists ) , pharmacology and clinical developments . The thienopyridines , ticlopidine and clopidogrel , are antiplatelet drugs . They are prodrugs and are metabolised in the liver to active metabolites that are non-competitive antagonists of the platelet adenosine diphosphate receptor , Q9H244 . Inhibition of platelet aggregation by these drugs is delayed until 24-48 h after administration , with maximal inhibition achieved after 3-5 days . Recovery of platelet function after drug withdrawal is slow ( 7-14 days ) . DB00208 and clopidogrel are effective in preventing atherothrombotic events in cardiovascular , cerebrovascular and peripheral vascular disease . Gastrointestinal side effects and skin rashes are common . However , neutropenia and thrombotic thrombocytopenic purpura are significant and sometimes fatal adverse effects of ticlopidine . DB00758 appears to offer several advantages over ticlopidine : a more rapid onset of action and a lower incidence of neutropenia and thrombotic thrombocytopenic purpura.A combination of clopidogrel and aspirin has become standard for antithrombotic therapy in cardiovascular disease . The anaesthetic considerations of patients taking the thienopyridine compounds are discussed . Pharmacological effects of lipid-lowering drugs recapitulate with a larger amplitude the phenotypic effects of common variants within their target genes . BACKGROUND : A major expectation underlying the search for novel susceptibility genes for common diseases using genome-wide association studies ( GWAS ) is that these discoveries will lead to new drug targets . This claim has not been verified yet . Here , we tested the hypothesis that common single nucleotide polymorphisms ( SNPs ) within drug target genes are associated with the corresponding phenotypes , using a population-based GWAS dataset and lipid-lowering drugs as a test case . METHODS : We examined the association between 36 genotyped and 193 imputed SNPs within four lipid-lowering drug target genes ( P04035 , Q07869 , Q8TDS4 / Q8TDS4 and P11597 ) and four non-lipid drug target genes ( P12821 , P30556 , Q9H244 , and P51164 ) and lipid phenotypes , blood pressure , and coronary artery disease in 5635 adult participants of the Lausanne , Switzerland , CoLaus study , genotyped using the Affymetrix 500K SNP chip technology . RESULTS : The phenotypes associated with SNPs within drug target genes recapitulated to a certain extent the pharmacological effects of the drug . The amplitude of the SNP effect was about 10 times smaller than the pharmacological effect of the corresponding drug . In particular , several P11597 SNPs were associated with an elevation in HDL-cholesterol levels , yet a lower diastolic blood pressure , providing evidence that the blood pressure elevation induced by the P11597 inhibitor torcetrapib is more likely compound specific than class specific . CONCLUSION : Pharmacological modulation of lipid-lowering drug targets recapitulates , and markedly amplifies , the phenotypic effects of common SNPs within these target genes . This data provides indirect evidence that , with certain limitations , large-scale GWAS represent a new tool for the discovery and the development of innovative drugs . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 and an inducible form termed P35354 . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 and P35354 . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 with the P23219 inhibitor and an R0 of 21 A for P35354 with the P35354 inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 and 18 A in P35354 . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Platelet Q9H244 receptor inhibition by thienopyridines : status and future . Thienopyridines have a well-established role in the treatment of coronary artery disease , especially in the setting of acute coronary syndromes and percutaneous coronary interventions . DB00208 , the first FDA-approved thienopyridine , was shown to be effective in reducing coronary events in high risk patients , but the original enthusiasm was hampered by concerns about its serious bone marrow toxicity . DB00758 a second generation thienopyridine with lesser side effects , is not only at least as effective as ticlopidine , but in combination with a low dose of aspirin , has been demonstrated to reduce the risk of major cardiovascular events in acute coronary syndrome patients in large-scale , randomised trials . Recent studies have highlighted major flaws in clopidogrel pharmacokinetics due to its delayed onset of action , and much attention has been devoted to the phenomenon of clopidogrel ' resistance ' . Among the novel , third generation thienopyridines , prasugrel as compared to clopidogrel has demonstrated lower inter-patient response variability and a reduced incidence of ischaemic events , but at an increased risk of major bleeding . Currently , several studies are continuing to test new direct Q9H244 receptor antagonists , such as cangrelor and AZD6140 , characterised by a faster reversal of platelet inhibition . Dehydration activates an NF-kappaB-driven , P35354 -dependent survival mechanism in renal medullary interstitial cells . Renal prostaglandin ( PG ) synthesis is mediated by cyclooxygenase-1 and -2 ( P23219 and P35354 ) . After dehydration , the maintenance of normal renal function becomes particularly dependent upon PG synthesis . The present studies were designed to examine the potential link between medullary P23219 and P35354 expression in hypertonic stress . In response to water deprivation , P35354 , but not P23219 , mRNA levels increase significantly in the renal medulla , specifically in renal medullary interstitial cells ( RMICs ) . DB09145 deprivation also increases renal NF-kappaB-driven reporter expression in transgenic mice . NF-kappaB activity and P35354 expression could be induced in cultured RMICs with hypertonic sodium chloride and mannitol , but not urea . RMIC P35354 expression was also induced by driving NF-kappaB activation with a constitutively active O15111 alpha ( IKKalpha ) . Conversely , introduction of a dominant-negative IkappaB mutant reduced P35354 expression after hypertonicity or IKKalpha induction . RMICs failed to survive hypertonicity when P35354 was downregulated using a P35354 -selective antisense or blocked with the selective nonsteroidal anti-inflammatory drug ( NSAID ) SC58236 , reagents that did not affect cell survival in isotonic media . In rabbits treated with SC58236 , water deprivation induced apoptosis of medullary interstitial cells in the renal papilla . These results demonstrate that water deprivation and hypertonicity activate NF-kappaB . The consequent increase in P35354 expression favors RMIC survival in hypertonic conditions . Inhibition of RMIC P35354 could contribute to NSAID-induced papillary injury . Mechanism of activation and functional role of protein kinase Ceta in human platelets . The novel class of protein kinase C ( nPKC ) isoform eta is expressed in platelets , but not much is known about its activation and function . In this study , we investigated the mechanism of activation and functional implications of nPKCeta using pharmacological and gene knock-out approaches . nPKCeta was phosphorylated ( at DB00156 -512 ) in a time- and concentration-dependent manner by 2MeSADP . Pretreatment of platelets with P59665 -2179 , a P47900 receptor antagonist , or YM-254890 , a G(q) blocker , abolished 2MeSADP-induced phosphorylation of nPKCeta . Similarly , ADP failed to activate nPKCeta in platelets isolated from P47900 and G(q) knock-out mice . However , pretreatment of platelets with Q9H244 receptor antagonist , AR-C69331MX did not interfere with ADP-induced nPKCeta phosphorylation . In addition , when platelets were activated with 2MeSADP under stirring conditions , although nPKCeta was phosphorylated within 30 s by ADP receptors , it was also dephosphorylated by activated integrin alpha(IIb)beta3 mediated outside-in signaling . Moreover , in the presence of SC-57101 , a alpha(IIb)beta3 receptor antagonist , nPKCeta dephosphorylation was inhibited . Furthermore , in murine platelets lacking PP1cgamma , a catalytic subunit of serine/threonine phosphatase , alpha(IIb)beta3 failed to dephosphorylate nPKCeta . Thus , we conclude that ADP activates nPKCeta via P47900 receptor and is subsequently dephosphorylated by PP1gamma phosphatase activated by alpha(IIb)beta3 integrin . In addition , pretreatment of platelets with eta-RACK antagonistic peptides , a specific inhibitor of nPKCeta , inhibited ADP-induced thromboxane generation . However , these peptides had no affect on ADP-induced aggregation when thromboxane generation was blocked . In summary , nPKCeta positively regulates agonist-induced thromboxane generation with no effects on platelet aggregation . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared .

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In vitro and in vivo activities of gatifloxacin against Mycobacterium tuberculosis . DB01044 ( Q6IB77 ) and moxifloxacin ( MXF ) were evaluated in vitro to determine their activities against Mycobacterium tuberculosis . Q6IB77 was subsequently compared in a dose range study to isoniazid ( DB00951 ) in a murine tuberculosis model . Q6IB77 was somewhat less active than DB00951 . Q6IB77 and MXF were evaluated in mice infected with M. tuberculosis and were found to have similar activities . Q6IB77 was studied alone and in combination with ethambutol , ethionamide ( P25101 ) , and pyrazinamide ( PZA ) and compared to DB00951 and rifampin ( Q9HBH0 ) . Q6IB77 appears to have sufficient activity alone and in combination with P25101 with or without PZA to merit evaluation for treatment of tuberculosis . P00797 -angiotensin system expression in rat bone marrow haematopoietic and stromal cells . The existence of a bone marrow renin-angiotensin system ( DB01367 ) is evidenced by the association of renin , angiotensin converting enzyme ( P12821 ) , and angiotensin ( Ang ) II and its AT(1) and AT(2) receptors with both normal and disturbed haematopoiesis . The expression of DB01367 components by rat unfractionated bone marrow cells ( BMC ) , haematopoietic-lineage BMC and cultured marrow stromal cells ( O60682 ) was investigated to determine which specific cell types may contribute to a local bone marrow DB01367 . The mRNAs for angiotensinogen , renin , P12821 , and AT(1a) and AT(2) receptors were present in BMC and in cultured O60682 ; Q9BYF1 mRNA was detected only in BMC . Two-colour flow fluorocytometry analysis showed immunodetectable angiotensinogen , P12821 , AT(1) and AT(2) receptors , and Ang II , as well as binding of Ang II to AT(1) and AT(2) receptors , in P01730 (+) , CD11b/c(+) , CD45R(+) and CD90(+) BMC and cultured O60682 ; renin was found in all cell types with the exception of P01730 (+) BMC . Furthermore , Ang II was detected by radioimmunoassay in O60682 homogenates as well as conditioned culture medium . The presence of Ang II receptors in both haematopoietic-lineage BMC and O60682 , and the de novo synthesis of Ang II by O60682 suggest a potential autocrine-paracrine mechanism for local DB01367 -mediated regulation of haematopoiesis . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . P27361 /2 activation modulates pyocyanin-induced toxicity in A549 respiratory epithelial cells . Pyocyanin ( Q15149 ) , a virulence factor produced by Pseudomonas aeruginosa , has many damaging effects on mammalian cells . Several lines of evidence suggest that this damage is primarily mediated by its ability to generate oxidative stress . However mechanisms underlying Q15149 -induced oxidative injury remain unclear . Although oxidative stress and subsequent MAPK signaling has been shown to modulate cell death in other models , its role in Q15149 -induced cytotoxicity remains unknown . Therefore the aim of this study was to investigate the role of redox-sensitive MAPK in Q15149 -induced toxicity in A549 cells . Here we show that Q15149 ( 50μM ) rapidly increased P27361 /2 phosphorylation after 5min . Pre-treatment of A549 cells with the Q02750 /2 inhibitor U0126 ( 10μM ) decreased Q15149 -induced P27361 /2 phosphorylation and protected cells against apoptosis and cell injury suggesting a role for P29323 signalling . In contrast , JNK and p38 MAPK phosphorylation remained unchanged following exposure to Q15149 and pretreatment with either the JNK or p38 MAPK inhibitors ( 10μM SP600125 and 10μM SB203580 , respectively ) did not afford protection against Q15149 toxicity . This would suggest that Q15149 -induced cytotoxicity appears to occur independently of JNK and p38 MAPK signaling pathways . Finally , although we confirm that oxidative stress contributes to Q15149 -induced toxicity , our data suggest the contribution of oxidative stress is independent of P27361 /2 signaling . These findings may provide insight for novel targeted therapies to reduce Q15149 -mediated lung injury in patients with chronic P. aeruginosa respiratory infections . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . Measuring ligand-dependent and ligand-independent interactions between nuclear receptors and associated proteins using Bioluminescence Resonance Energy Transfer ( BRET ) . Bioluminescent resonance energy transfer ( BRET2 ) is a recently developed technology for the measurement of protein-protein interactions in a live , cell-based system . BRET2 is characterized by the efficient transfer of excited energy between a bioluminescent donor molecule ( Renilla luciferase ) and a fluorescent acceptor molecule ( a mutant of Green Fluorescent Protein ( GFP2 ) . The BRET2 assay offers advantages over fluorescence resonance energy transfer ( FRET ) because it does not require an external light source thereby eliminating problems of photobleaching and autoflourescence . The absence of contamination by light results in low background that permits detection of very small changes in the BRET2 signal . BRET2 is dependent on the orientation and distance between two fusion proteins and therefore requires extensive preliminary standardization experiments to conclude a positive BRET2 signal independent of variations in protein titrations and arrangement in tertiary structures . P03372 ( ER ) signaling is modulated by steroid receptor coactivator 1 ( Q15788 ) . To establish BRET2 in a ligand inducible system we used Q15788 as the donor moiety and ER as the acceptor moiety . Expression and functionality of the fusion proteins were assessed by transient transfection in P29320 -293 cells followed by Western blot analysis and measurement of ER-dependent reporter gene activity . These preliminary determinations are required prior to measuring nuclear receptor protein-protein interactions by BRET2 . This article describes in detail the BRET2 methodology for measuring interaction between full-length ER and coregulator proteins in real-time , in an in vivo environment . Integrative analysis of proteomic and transcriptomic data for identification of pathways related to simvastatin-induced hepatotoxicity . Hepatocytes are used widely as a cell model for investigation of xenobiotic metabolism and the toxic mechanism of drugs . Simvastatin is the first statin drug used extensively in clinical practice for control of elevated cholesterol or hypercholesterolemia . However , it has also been reported to cause adverse effects in liver due to cellular damage . In this study , for proteomic and transcriptomic analysis , rat primary hepatocytes were exposed to simvastatin at IC20 concentration for 24 h . Among a total of 607 differentially expressed proteins , 61 upregulated and 29 downregulated proteins have been identified in the simvastatin-treated group . At the mRNA level , results of transcriptomic analysis revealed 206 upregulated and 41 downregulated genes in the simvastatin-treated group . Based on results of transcriptomic and proteomic analysis , Q16236 -mediated oxidative stress response , xenobiotics by metabolism of cytochrome P450 , fatty acid metabolism , bile metabolism , and urea cycle and inflammation metabolism pathways were focused using IPA software . Genes ( P49327 , UGT2B , P00352 , P05177 , P09210 , HAP90 , P05231 , IL-1 , P15090 , and ABC11 ) and proteins ( P49327 , CYP2D1 , UG2TB , P00352 , P09210 , HSP90 , P15090 , and O95342 ) related to several important pathways were confirmed by real-time PCR andWestern blot analysis , respectively . This study will provide new insight into the potential toxic pathways induced by simvastatin . Evidence of drug-drug interactions through uptake and efflux transport systems in rat hepatocytes : implications for cellular concentrations of competing drugs . For drugs with hepatobiliary transport across hepatocytes , the interplay between uptake and efflux transporters determines hepatic concentrations of drugs , but the evolution over time of these concentrations is difficult to measure in humans other than with magnetic resonance imaging contrast agents in the liver . DB00743 dimeglumine ( BOPTA ) is a contrast agent used in liver magnetic resonance imaging that enters into human hepatocytes through organic anion transporting polypeptides ( P46721 ) and exits unchanged into bile through the multiple resistance-associated protein 2 ( Q92887 ) . DB01045 ( Q9HBH0 ) is transported by the same membrane proteins and may compete with BOPTA for hepatic uptake . Simultaneous drug-drug interactions through uptake and efflux transport systems in hepatocytes according to the cellular concentrations of competing drugs were never investigated . In perfused rat liver preparations , we demonstrate how the drug-drug interactions through transporters determine cellular concentrations of the competing drugs BOPTA and Q9HBH0 , and we show that the cellular concentrations by modulating transport through membranes regulate the rat Oatp-Mrp2 interplay . Moreover , drug interactions through transporters change greatly over time . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . DB01045 -independent interactions between the pregnane X receptor ligand binding domain and peptide fragments of coactivator and corepressor proteins . The pregnane X receptor ( O75469 ) , a member of the nuclear receptor superfamily , regulates the expression of drug-metabolizing enzymes in a ligand-dependent manner . The conventional view of nuclear receptor action is that ligand binding enhances the receptor 's affinity for coactivator proteins , while decreasing its affinity for corepressors . To date , however , no known rigorous biophysical studies have been conducted to investigate the interaction among O75469 , its coregulators , and ligands . In this work , steady-state total internal reflection fluorescence microscopy ( TIRFM ) and total internal reflection with fluorescence recovery after photobleaching were used to measure the thermodynamics and kinetics of the interaction between the O75469 ligand binding domain and a peptide fragment of the steroid receptor coactivator-1 ( Q15788 ) in the presence and absence of the established O75469 agonist , rifampicin . Equilibrium dissociation and dissociation rate constants of ~5 μM and ~2 s(-1) , respectively , were obtained in the presence and absence of rifampicin , indicating that the ligand does not enhance the affinity of the O75469 and Q15788 fragments . Additionally , TIRFM was used to examine the interaction between O75469 and a peptide fragment of the corepressor protein , the silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) . An equilibrium dissociation constant of ~70 μM was obtained for Q9Y618 in the presence and absence of rifampicin . These results strongly suggest that the mechanism of ligand-dependent activation in O75469 differs significantly from that seen in many other nuclear receptors . O75469 induces Q02318 and regulates cholesterol metabolism in the intestine . Mitochondrial sterol 27-hydroxylase ( Q02318 ) catalyzes oxidative cleavage of the sterol side chain in the bile acid biosynthetic pathway in the liver and 27-hydroxylation of cholesterol in most tissues . Recent studies suggest that 27-hydroxycholesterol ( 27-HOC ) activates liver orphan receptor alpha ( LXRalpha ) and induces the cholesterol efflux transporters O95477 and P45844 in macrophages . The steroid- and bile acid-activated pregnane X receptor ( O75469 ) plays critical roles in the detoxification of bile acids , cholesterol metabolites , and xenobiotics . The role of Q02318 in the intestine is not known . This study investigated O75469 and Q02318 regulation of cholesterol metabolism in the human intestinal cell lines Caco2 and Ls174T . A human O75469 ligand , rifampicin , induced Q02318 mRNA expression in intestine cells but not in liver cells . DB01045 induced Q02318 gene transcription , increased intracellular 27-HOC levels , and induced O95477 and P45844 mRNA expression only in intestine cells . A functional O75469 binding site was identified in the human Q02318 gene . Chromatin immunoprecipitation assays revealed that rifampicin induced the O75469 recruitment of steroid receptor coactivator 1 to Q02318 chromatin . DB04540 loading markedly increased intracellular 27-HOC levels in intestine cells . DB01045 , 27-HOC , and a potent LXRalpha agonist , T0901317 , induced O95477 and P45844 protein expression and stimulated cholesterol efflux from intestine cells to apolipoprotein A-I and HDL . This study suggests an intestine-specific O75469 / Q02318 /LXRalpha pathway that regulates intestine cholesterol efflux and HDL assembly . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . The steroid receptor co-activator-1 ( Q15788 ) potentiates TGF-beta/Smad signaling : role of p300/CBP . The three related 160-kDa proteins , Q15788 , Q06418 -2 and Q9Y6Q9 , were initially identified as factors interacting with nuclear receptors . They have also been reported to potentiate the activity of other transcription factors such as AP-1 or NF-kappaB . The aim of this work was to identify whether Q15788 interferes with the TGF-beta/Smad signaling pathway , and if so , to identify its underlying mechanisms of action . Using transient cell transfection experiments performed in human dermal fibroblasts with the P84022 /4-specific (SBE)4-lux reporter construct , as well as the human P05121 promoter , we determined that Q15788 enhances TGF-beta-induced , Smad-mediated , transcription . Likewise , Q15788 overexpression potentiated TGF-beta-induced upregulation of P05121 steady-state mRNA levels . Using a mammalian two-hybrid system , we demonstrated that Q15788 interacts with the transcriptional co-activators p300/CBP , but not with P84022 . Overexpression of the adenovirus E1A oncoprotein , an inhibitor of CBP/p300 activity , prevented the enhancing effect of Q15788 on P84022 /4-mediated transcription , indicating that p300/CBP may be required for Q15788 effect . Such hypothesis was validated , as expression of a mutant form of Q15788 lacking the CBP/p300-binding site failed to upregulate P84022 /4-dependent transcription , while full-length Q15788 potentiated p300. P84022 interactions . These results identify Q15788 as a novel P84022 /4 transcriptional partner , facilitating the functional link between P84022 and p300/CBP . Identification of macrophage genes responsive to extracellular acidification . OBJECTIVE : A low pH microenvironment is a characteristic feature of inflammation loci and affects the functions of immune cells . In this study , we investigated the effect of extracellular acidification on macrophage gene expression . METHODS : RAW264.7 macrophages were incubated in neutral ( pH 7.4 ) or acidic ( pH 6.8 ) medium for 4 h . Global mRNA expression levels were determined using Affymetrix genechips . RESULTS : The mRNA expressions of 353 macrophage genes were significantly modified after incubation in acidic medium ; 193 were up-regulated and 160 down-regulated . Differentially regulated genes were grouped into 13 classes based on the functions of the corresponding protein products . Pathway analysis revealed that differentially expressed genes are enriched in pathways related to inflammation and immune responses . Quantitative real-time PCR analysis confirmed that the expressions of P02778 , O95715 , Q14116 , P24394 , O95477 , P13236 , IL-7R , P61073 , Q9NYK1 , and P10147 mRNAs were regulated by extracellular acidification . CONCLUSION : The results of this study provide insights into the effects of acidic extracellular environments on macrophage gene expression . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . DB00640 A2A receptor : a target for regulating renal interstitial fibrosis in obstructive nephropathy . Renal interstitial fibrosis ( Q9HBH0 ) is the common pathological process of chronic kidney diseases leading inevitably to renal function deterioration . Q9HBH0 and its preceding epithelial-mesenchymal transition ( EMT ) are commonly triggered by an early occurring renal inflammation . However , an effective approach to prevent EMT and Q9HBH0 is still lacking and of urgent need . Recently , the adenosine A2A receptor ( A2AR ) emerges as a novel inflammation regulator , therefore manipulation of A2AR may suppress the EMT process and as such protect against Q9HBH0 . To test this hypothesis we applied a unilateral ureteral obstruction ( UUO ) model of Q9HBH0 on A2AR knockout mice and their wild-type littermates , combined with the intervention of a selective A2AR agonist , CGS 21680 . On days 3 , 7 and 14 post-UUO we evaluated the effects of A2AR manipulation on the molecular pathological progresses of Q9HBH0 , including the cellular component of interstitial infiltration , expression of profibrotic factors , cellular biomarkers of EMT , and collagen deposition of extracellular matrix . Our data demonstrated that activation of A2AR significantly suppressed the deposition of collagen types I and III , reduced the infiltration of P01730 + T lymphocytes , and attenuated the expression of TGF-β1 and Q13464 , which in turn inhibited and postponed the EMT progress . Conversely , genetic inactivation of A2AR exacerbated the aforementioned pathological processes of UUO-induced Q9HBH0 . Together , activation of A2AR effectively alleviated EMT and Q9HBH0 in mice , suggesting A2AR as a potential therapeutic target for the treatment of Q9HBH0 . Organic anion transporting polypeptide-C mediates arsenic uptake in P29320 -293 cells . Arsenic is an established human carcinogen . The role of aquaglyroporins ( AQPs ) in arsenic disposition was recently identified . In order to examine whether organic anion transporting polypeptide-C ( Q9Y6L6 ) also plays a role in arsenic transport , Q9Y6L6 cDNA was transfected into cells of a human embryonic kidney cell line ( P29320 -293 ) . Transfection increased uptake of the model Q9Y6L6 substrate , estradiol-17beta-D-glucuronide , by 10-fold . In addition , we measured uptake and cytotoxicity of arsenate , arsenite , monomethylarsonate(MMA(V)) , and dimethylarsinate ( P28067 (V) ) . Transfection of Q9Y6L6 increased uptake and cytotoxicity of arsenate and arsenite , but not of MMA(V) or P28067 (V) . DB01045 and taurocholic acid ( a substrate of Q9Y6L6 ) reversed the increased toxicity of arsenate and arsenite seen in Q9Y6L6 -transfected cells . The increase in uptake of inorganic arsenic was not as great as that of estradiol-17beta-D-glucuronide . Our results suggest that Q9Y6L6 can transport inorganic arsenic in a ( DB00143 ) -dependent manner . However , this may not be the major pathway for arsenic transport . P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 -binding cassette transporter A1 ( O95477 ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 directly binds calmodulin in a Ca(2+)-dependent manner . The cytoplasmic loop of O95477 contains a typical calmodulin binding sequence of 1-5-8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5-8-14 motif abolished this interaction . This motif is located near the O95477 Pro- DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin/Ca(2+) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 and decreased O95477 protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 in the presence of Ca(2+) and increases the generation of high-density lipoprotein . Rapid detection of multidrug-resistant Mycobacterium tuberculosis by use of real-time PCR and high-resolution melt analysis . The current study describes the development of a unique real-time PCR assay for the detection of mutations conferring drug resistance in Mycobacterium tuberculosis . The rifampicin resistance determinant region ( RRDR ) of rpoB and specific regions of katG and the inhA promoter were targeted for the detection of rifampin ( Q9HBH0 ) and isoniazid ( DB00951 ) resistance , respectively . Additionally , this assay was multiplexed to discriminate Mycobacterium tuberculosis complex ( P04629 ) strains from nontuberculous Mycobacteria ( Q9P121 ) strains by targeting the IS6110 insertion element . High-resolution melting ( HRM ) analysis following real-time PCR was used to identify M. tuberculosis strains containing mutations at the targeted loci , and locked nucleic acid ( LNA ) probes were used to enhance the detection of strains containing specific single-nucleotide polymorphism ( SNP ) transversion mutations . This method was used to screen 252 M. tuberculosis clinical isolates , including 154 Q9HBH0 -resistant strains and 174 DB00951 -resistant strains based on the agar proportion method of drug susceptibility testing ( Q03001 ) . Of the 154 Q9HBH0 -resistant strains , 148 were also resistant to DB00951 and therefore classified as multidrug resistant ( MDR ) . The assay demonstrated sensitivity and specificity of 91 % and 98 % , respectively , for the detection of Q9HBH0 resistance and 87 % and 100 % for the detection of DB00951 resistance . Overall , this assay showed a sensitivity of 85 % and a specificity of 98 % for the detection of MDR strains . This method provides a rapid , robust , and inexpensive way to detect the dominant mutations known to confer MDR in M. tuberculosis strains and offers several advantages over current molecular and culture-based techniques . Inhibition of a thrombin anion-binding exosite-2 mutant by the glycosaminoglycan-dependent serpins protein C inhibitor and heparin cofactor II . Antithrombin ( P01008 ) , heparin cofactor II ( HCII ) and protein C inhibitor ( P05154 ; also named plasminogen activator inhibitor-3 ) are serine protease inhibitors ( serpins ) whose thrombin inhibition activity is accelerated in the presence of glycosaminoglycans . We compared the inhibition properties of P05154 and HCII to P01008 using R93A/R97A/R101A thrombin , an anion-binding exosite-2 ( exosite-2 ) mutant that has greatly reduced heparin-binding properties . DB01109 -enhanced P05154 inhibition of R93A/R97A/R101A thrombin was only approximately 2-fold compared to 40-fold enhancement with wild-type recombinant thrombin . P07204 ( TM ) ( with or without the chondroitin sulfate moiety ) accelerated P05154 inhibition of both wild-type and R93A/R97A/R101A thrombins . HCII achieved the same maximum activity in the presence of heparin with both wild-type and R93A/R97A/R101A thrombins ; however , the optimum heparin concentration was 20 times greater than the reaction with wild-type thrombin , indicative of a decrease in heparin affinity . Dermatan sulfate ( DSO4 ) -catalyzed HCII thrombin inhibition was unchanged in R93A/R97A/R101A thrombin compared to wild-type recombinant thrombin . These results suggest that P05154 is similar to P01008 and depends upon ternary complex formation with heparin and these specific thrombin exosite-2 residues to accelerate thrombin inhibition . In contrast , HCII does not require DB00125 (93) , DB00125 (97) and DB00125 (101) of thrombin exosite-2 and further supports the hypothesis that HCII uses an allosteric process following glycosaminoglycan binding to inhibit thrombin . A Q8NBP7 -binding antibody that structurally mimics the P01133 (A) domain of LDL-receptor reduces LDL cholesterol in vivo . Proprotein convertase subtilisin-like/kexin type 9 ( Q8NBP7 ) regulates LDL cholesterol levels by inhibiting P01130 ( LDLr ) -mediated cellular LDL uptake . We have identified a fragment antigen-binding ( Fab ) 1D05 which binds Q8NBP7 with nanomolar affinity . The fully human antibody 1D05-IgG2 completely blocks the inhibitory effects of wild-type Q8NBP7 and two gain-of-function human Q8NBP7 mutants , S127R and D374Y . The crystal structure of 1D05-Fab bound to Q8NBP7 reveals that 1D05-Fab binds to an epitope on the Q8NBP7 catalytic domain which includes the entire LDLr P01133 (A) binding site . Notably , the 1D05-Fab CDR-H3 and CDR-H2 loops structurally mimic the P01133 (A) domain of LDLr . In a transgenic mouse model ( P11597 /LDLr-hemi ) , in which plasma lipid and Q8NBP7 profiles are comparable to those of humans , 1D05-IgG2 reduces plasma LDL cholesterol to 40 % and raises hepatic LDLr protein levels approximately fivefold . Similarly , in healthy rhesus monkeys , 1D05-IgG2 effectively reduced LDL cholesterol 20 % -50 % for over 2 weeks , despite its relatively short terminal half-life ( t(1/2) = 3.2 days ) . Importantly , the decrease in circulating LDL cholesterol corresponds closely to the reduction in free Q8NBP7 levels . Together these results clearly demonstrate that the LDL-lowering effect of the neutralizing anti- Q8NBP7 1D05-IgG2 antibody is mediated by reducing the amount of Q8NBP7 that can bind to the LDLr . Vertical profile of dioxin-like and estrogenic potencies in a sediment core from Tokyo Bay , Japan . Dioxin-like and estrogenic activities were measured in a sediment core collected from Tokyo Bay using in vitro bioassays after fractionating sediment extracts into three fractions by florisil column chromatography . Target analytes including polychlorinated biphenyls ( PCBs ) , polychlorinated naphthalenes ( PCNs ) , polycyclic aromatic hydrocarbons ( PAHs ) and nonylphenol ( NP ) were measured by gas chromatography-mass spectrometry ( GC-MS ) or high performance liquid chromatography ( HPLC ) techniques . NP concentrations were greater in surface sediments ( 0 to 12 cm ) than those in sub-surface ( 12-30 cm ) . The maximum observed Q15149 concentration was 5 ng/g , whereas that of PCBs was 300 ng/g . Concentrations of PCBs in fraction 1 ( F1 ) were not high enough to induce significant luciferase activities in H4IIE-luc cells . Significant dioxin-like activities were found in fractions 2 ( F2 ) and 3 ( P13726 ) . PAH concentrations were correlated with dioxin-like activities measured in F2 . Compounds that contribute to the dioxin-like activities in P13726 were not identified yet . Significant estrogenic activities were observed in F2 samples , which may be related to the presence of certain estrogenic PAHs . P13726 samples were cytotoxic to MCF-7 cells and therefore their estrogenic potential could not be estimated . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Attenuation of anti-tuberculosis therapy induced hepatotoxicity by Spirulina fusiformis , a candidate food supplement . Therapy using Isoniazid ( DB00951 ) and DB01045 ( Q9HBH0 ) leads to induction of hepatotoxicity in some individuals undergoing anti-tuberculosis treatment . In this study , we assessed the effect of Spirulina fusiformis on DB00951 and Q9HBH0 induced hepatotoxicity in rats compared with hepatoprotective drug Silymarin . Induction of hepatotoxicity was measured by changes in the liver marker enzymes ( aspartate transaminase , alanine transaminase , and alkaline phosphatase ) . The antioxidant status was also analyzed in liver tissue homogenate and plasma by measurement of superoxide dismutase , catalase , glutathione-S-transferase , glutathione reductase , and lipid peroxidation levels . We also aimed to study the binding and interactions of the transcription factors Pregnane X Receptor ( O75469 ) and Farnesoid X Receptor ( Q96RI1 ) with DB00951 , Q9HBH0 , and representative active compounds of Spirulina fusiformis by in silico methods . The administration of DB00951 and Q9HBH0 resulted in significant ( p < 0.05 ) decrease in the antioxidant levels and total protein levels . There was also a significant ( p < 0.05 ) increase in the levels of liver marker enzymes . Spirulina fusiformis was seen to protect the parameters from significant changes upon challenge with DB00951 and Q9HBH0 in a dose-dependent manner . This was corroborated by histological examination of the liver . The results of the in silico analyses further support the wet lab results . Anti-atherosclerotic effects of sirolimus on human vascular smooth muscle cells . DB00877 is a potent immunosuppressive agent and has an anti-atherosclerotic effect through its anti-proliferative property . The present study was undertaken to investigate the effect of sirolimus on intracellular cholesterol homeostasis in human vascular smooth muscle cells ( VSMCs ) in the presence of inflammatory cytokine P01584 . We explored the effect of sirolimus on the lipid accumulation of VSMCs in the presence of P01584 , using Oil Red O staining and quantitative measurement of intracellular cholesterol . The effect of sirolimus on the gene and protein expression of lipoprotein receptors and DB00171 binding cassettes ( O95477 and P45844 ) was examined by real-time PCR and Western blotting , respectively . Furthermore , the effect of sirolimus on cholesterol efflux from VSMCs in the presence or absence of P01584 was also investigated using [ (3)H ] cholesterol efflux . Finally , we examined the effect of sirolimus on the production of inflammatory cytokines in VSMCs using ELISA . DB00877 reduced intracellular lipid accumulation in VSMCs mediated by P01584 possibly due to the reduction of expression of low-density lipoprotein ( LDL ) and very low-density lipoprotein ( VLDL ) receptors . DB00877 increased cholesterol efflux from VSMCs and overrode the suppression of cholesterol efflux induced by P01584 . DB00877 also increased O95477 and P45844 genes expression , even in the presence of P01584 . We further confirmed that sirolimus inhibited mRNA and protein expression of inflammatory cytokines P05231 , tumor necrosis factor-alpha , P10145 , and monocyte chemoattractant protein-1 . Inhibition of lipid uptake together with increasing cholesterol efflux and the inhibition of inflammatory cytokines are all important aspects of the anti-atherosclerotic effects of sirolimus on VSMCs . Genetic aspects of ischemic stroke : coagulation , homocysteine , and lipoprotein metabolism as potential risk factors . Stroke is one of the most common causes of death and long term disability throughout the world . It may be the outcome of a number of monogenic disorders or , more commonly , a polygenic multifactorial disease . Numerous studies have investigated the role of genetics in the pathogenesis of ischemic stroke , with varied and often contradictory results . The candidate ' stroke risk ' genes affecting haemostasis ( P12259 , F2 , P02671 / P02675 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / P08514 , P17301 , P07359 , TPA , Q96IY4 , P07204 , PZ , P08758 ) , homocysteine metabolism ( P42898 , P35520 , Q99707 ) , and lipid metabolism ( apo E , P06858 , P11597 , O95477 , apo AI , apo CIII , apo AIV , apo AV , apo B , apo H , apo(a) , P27169 /2/3 , P01130 / P78380 ) are evaluated in this review . By examining meta-analyses and case-control studies , we made a classification of gene/gene polymorphisms according to the degree of association with ischemic stroke risk . The data assembled could be very useful for further meta-analysis and for future clinical applications . Steroid hormone receptors and coregulators in endocrine-resistant and estrogen-independent breast cancer cells . Resistance to hormonal therapy is often a problem in the treatment of breast cancer patients . It has been suggested that resistance could be explained by altered nuclear hormone receptor or coregulator levels or inappropriately increased agonist activity of selective estrogen receptor modulator ( SERM ) . To test these hypotheses , we have established novel MCF-7 cell line-derived in vitro models of anti-estrogen- and progestin-resistant and estrogen-independent breast cancer by long-term culture in the presence of toremifene and medroxyprogesterone acetate ( DB00603 ) and in the absence of estradiol , respectively . Using cell growth and multiprobe ribonuclease protection assays , the expression of 5 nuclear hormone receptors and 9 coregulators as well as the alterations in the cell proliferation and target gene transcription in response to hormonal treatments were studied . P06401 ( PR ) expression was decreased and silencing mediator for retinoid acid and thyroid hormone receptors ( Q9Y618 ) and amplified in breast cancer-1 ( Q9Y6Q9 ) expression increased in anti-estrogen-resistant cells . Estrogen caused PR and ERbeta upregulation in all cell lines , but we did not observe increased agonist activity of anti-estrogen measured by regulation of these estrogen target genes . Basal ERalpha levels and estrogenic growth response were decreased and p300/CBP-associated factor ( pCAF ) and Q9Y6Q9 upregulated by estrogen in progestin-resistant cells , but coregulator levels were unchanged . Estrogen-independent cells were still estrogen-responsive and PR , nuclear receptor corepressor ( O75376 ) and Q9Y618 expression was increased whereas steroid receptor coactivator-1 ( P12931 -1a ) and CBP-related protein p300 ( p300 ) expression decreased . Their growth was inhibited by toremifene , but estradiol was able to abrogate this effect , which might have interesting clinical implications concerning the use of postmenopausal hormone replacement therapy . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Bresol inhibits phosphodiesterase 4 gene expression and modulates the levels of select mediators of inflammation in human monocytic cells . Bresol-a poly-herbal formulation , has been reported to be effective against bronchial asthma and allergic rhinitis in children . In vivo studies have supported the anti-histaminic and anti-anaphylactic action of bresol . However , the mechanism of action of bresol in modulation of inflammation has not been studied at the cellular and molecular level . The present study was aimed to elucidate the mechanism(s) of action of bresol at the cellular and molecular levels , using human monocyte leukemia cells . The effects of bresol on phosphodiesterase 4B ( Q07343 ) gene expression were analyzed using human monocytic U937 leukemia cells . The ability of bresol to stimulate DB02527 formation in these cells , as well as its effects on mediators of inflammation like tumor necrosis factor-α ( TNFα ) , nitric oxide ( NO ) , and cycloxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated U937 cells , were also studied . The results here indicated that bresol exhibited potential anti-inflammatory properties by inhibiting LPS-induced Q07343 gene expression in the cells . Bresol also dose dependently activated DB02527 formation , and inhibited TNFα , NO , as well as P35354 formation in the LPS-stimulated cells . Based upon the results , we concluded that the reported anti-inflammatory activity of bresol might be attributed to its abilities to inhibit Q07343 and thus elevate DB02527 levels in human monocytes . The anti-inflammatory effects of bresol might also be a result of the capacity of bresol to modulate the formation of TNFα , NO , and P35354 in monocytes . Effects of usnic acid exposure on human hepatoblastoma HepG2 cells in culture . Usnic acid , a natural botanical product , is a constituent of some dietary supplements used for weight loss . It has been associated with clinical hepatotoxicity leading to liver failure in humans . The present study was undertaken for metabolism and toxicity evaluations of usnic acid in human hepatoblastoma HepG2 cells in culture . The cells were treated with the vehicle control and usnic acid at concentrations of 0-100 µm for 24 h at 37 °C in 5 % CO2 . Following the treatment period , the cells were evaluated by biochemical and toxicogenomic endpoints of toxicity that included cytochrome P450 activity , cytotoxicity , oxidative stress , mitochondrial dysfunction and changes in pathway focused gene expression profiles . Usnic acid exposure resulted in increased P450 activity , cytotoxicity , oxidative stress and mitochondrial dysfunction in HepG2 cells . The pathway-focused gene expression analysis resulted in significantly altered expression of six genes out of a total of 84 genes examined . Of the six altered genes , three genes were up-regulated and three genes down-regulated . A marked up-regulation of one gene O00585 associated with inflammation , one gene P24863 associated with proliferation and carcinogenesis and one gene P22310 associated with metabolism as well as DNA damage and repair were observed in the usnic acid-treated cells compared with the vehicle control . Also a marked down-regulation of one gene P04141 associated with inflammation and two genes ( P22680 and P05181 ) associated with oxidative metabolic stress were observed in the usnic acid-treated cells compared with the control . The biomarkers used in this study demonstrate the toxicity of usnic acid in human hepatoblastoma HepG2 cells , suggesting an oxidative mechanism of action . P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . Regulation of Q9BYW2 {alpha} activity in adipose tissue by obesity-associated factors : adipogenesis , insulin , and hypoxia . The transcription factor HIF-1α activity is increased in adipose tissue to contribute to chronic inflammation in obesity . However , its upstream and downstream events remain to be characterized in adipose tissue in obesity . We addressed this issue by investigating adipocyte HIF-1α activity in response to obesity-associated factors , such as adipogenesis , insulin , and hypoxia . In adipose tissue , both HIF-1α mRNA and protein were increased by obesity . The underlying mechanism was investigated in 3T3- Q9NUQ9 adipocytes . HIF-1α mRNA and protein were augmented by adipocyte differentiation . In differentiated adipocytes , insulin further enhanced HIF-1α in both levels . Hypoxia enhanced only HIF-1α protein , not mRNA . PI3K and P42345 activities are required for the HIF-1α expression . Function of HIF-1α protein was investigated in the regulation of P15692 gene transcription . ChIP assay shows that HIF-1α binds to the proximal hypoxia response element in the P15692 gene promoter , and its function is inhibited by a corepressor composed of O15379 and Q9Y618 . These observations suggest that of the three obesity-associated factors , all of them are able to augment HIF-1α protein levels , but only two ( adipogenesis and insulin ) are able to enhance HIF-1α mRNA activity . Adipose tissue HIF-1α activity is influenced by multiple signals , including adipogenesis , insulin , and hypoxia in obesity . The transcriptional activity of HIF-1α is inhibited by O15379 - Q9Y618 corepressor in the P15692 gene promoter . P35354 inhibitor treatment enhances photodynamic therapy-mediated tumor response . Photodynamic therapy ( PDT ) continues to be used in the treatment of solid tumors . Clinical results are promising , but the therapy has not been optimized , and tumor recurrences can occur . Recently , it has been shown that inhibitors of cyclooxygenase ( P36551 ) -2 can be effective in combination with conventional chemotherapy and radiation therapy . In the current study , we examined the parameters of PDT-mediated activation of P35354 expression . We also examined the tumoricidal effectiveness of combining PDT with the selective P35354 inhibitor NS-398 . PDT induced the transcriptional activation of P35354 . Prolonged expression of P35354 protein was observed in PDT-treated mouse sarcoma and carcinoma cell lines , whereas P23219 was not inducible by PDT . Prostaglandin ( PG ) E2 synthesis was also increased in PDT-treated cells , and DB00917 levels were attenuated in cells coincubated with NS-398 , indicating that PDT induced the expression of biologically active P35354 . Both porphyrin- and chlorin-based photosensitizers were able to elicit PDT-mediated P35354 expression . P35354 was also elevated in radiation-induced fibrosarcoma ( Q9HBH0 ) tumors after treatment with PDT . We also observed that systemic administration of NS-398 decreased PDT induction of both DB00917 and vascular endothelial growth factor in treated Q9HBH0 tumors . Additionally , we demonstrated that NS-398 enhanced PDT responsiveness in Q9HBH0 tumors without increasing toxicity to normal tissue . These results provide strong evidence that combination procedures involving selective P35354 inhibitors may improve the therapeutic effectiveness of PDT . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . The murine chemokine receptor P61073 is tightly regulated during T cell development and activation . We have characterized the murine homolog of the HIV-co-receptor P61073 during T cell development and activation . Our data demonstrate that this chemokine receptor , although highly conserved between human and mouse , is differently expressed and regulated in both species . Mitogenic activation resulted in an increase of surface P61073 on murine T cells within 2 days , whereas the receptor was strongly down-regulated on human T cells during this period . Furthermore , intraperitoneal immunization of mice resulted in a strong increase of splenic and mesenteric cytotoxic T cells co-expressing P61073 . It is interesting that , on thymocytes , expression of P61073 is restricted to P01730 +CD8+ cells . Stromal cell-derived factor-1alpha , a natural ligand of P61073 , induced chemotaxis of thymocytes and was found to counteract dexamethasone-induced apoptosis to a certain extent in these cells . Thus , our data show that expression of P61073 is tightly controlled on murine T cells and indicate that this highly conserved chemokine receptor might serve different functions in humans and mice . DNA-based prenatal diagnosis of plectin-deficient epidermolysis bullosa simplex associated with pyloric atresia . BACKGROUND : Mutations in the plectin gene ( Q15149 ) generally lead to epidermolysis bullosa simplex ( Q9BTE0 ) associated with muscular dystrophy . It has been recently demonstrated that Q15149 mutations can also cause a different clinical subtype , Q9BTE0 associated with pyloric atresia ( Q9BTE0 -PA ) , which shows early lethality . Prenatal diagnosis ( P01160 ) of Q9BTE0 -PA using mutation screening of Q15149 has not been described . OBJECTIVE : This study aimed to perform DNA-based P01160 for an Q9BTE0 -PA family . MATERIALS AND METHODS : The Q9BTE0 -PA proband was compound-heterozygous for a paternal c.1350G > A splice-site mutation and a maternal p.Q305X nonsense mutation . Genomic DNA was obtained from amniocytes taken from an at-risk fetus of the proband 's family . Direct sequencing and restriction enzyme digestion of polymerase chain reaction products from the genomic DNA were performed . RESULTS : Mutational analysis showed that the fetus harbored both pathogenic mutations , suggesting that the fetus was a compound-heterozygote and therefore affected with Q9BTE0 -PA . The skin sample obtained by autopsy from the abortus confirmed the absence of plectin expression at the dermal-epidermal junction . CONCLUSIONS : This is the first successful DNA-based P01160 for an EBA-PA family . DB11320 stimulates hydrogen peroxide production by bronchial epithelial cells via histamine H1 receptor and dual oxidase . Oxidative stress has been implicated in the pathogenesis of bronchial asthma . Besides granulocytes , the airway epithelium can produce large amounts of reactive oxygen species and can contribute to asthma-related oxidative stress . DB11320 is a major inflammatory mediator present in large quantities in asthmatic airways . Whether histamine triggers epithelium-derived oxidative stress is unknown . We therefore aimed at characterizing human airway epithelial H2O2 production stimulated by histamine . We found that air-liquid interface cultures of primary human bronchial epithelial cells ( BECs ) and an immortalized BEC model ( Cdk4/hTERT HBEC ) produce H2O2 in response to histamine . The main source of airway epithelial H2O2 is an NADPH dual oxidase , Duox1 . Out of the four histamine receptors ( P35367 - Q9H3N8 ) , P35367 has the highest expression in BECs and mediates the H2O2-producing effects of histamine . P05112 induces Duox1 gene and protein expression levels and enhances histamine-induced H2O2 production by epithelial cells . Using P29320 -293 cells expressing Duox1 or Duox2 and endogenous P35367 , histamine triggers an immediate intracellular calcium signal and H2O2 release . Overexpression of P35367 further increases the oxidative output of Duox-expressing P29320 -293 cells . Our observations show that BECs respond to histamine with Duox-mediated H2O2 production . These findings reveal a mechanism that could be an important contributor to oxidative stress characteristic of asthmatic airways , suggesting novel therapeutic targets for treating asthmatic airway disease . Effect of prototypical inducing agents on P-glycoprotein and CYP3A expression in mouse tissues . P-glycoprotein ( P-gp ) and CYP3A have considerable overlap in inducers in vitro . Characterizing P-gp induction in vivo and potential coregulation with CYP3A are important goals for predicting drug interactions . This study examined P-gp expression in mouse tissues and potential coinduction with CYP3A following oral treatment with 1 of 7 prototypical inducing agents for 5 days . P-gp expression in brain or liver was not induced by any treatment as determined by Western blot , whereas dexamethasone , pregnenolone-16alpha-carbonitrile ( Q15149 ) , St . John 's wort ( SJW ) , and rifampin induced hepatic CYP3A expression . In intestine , rifampin and SJW induced P-gp expression 3.7- and 1.6-fold and CYP3A 3.5- and 2.4-fold , respectively , whereas dexamethasone and Q15149 induced CYP3A only . These observations suggest that P-gp in mouse small intestine is inducible by some , but not all , CYP3A inducers , whereas P-gp expression in liver or brain is not readily induced . Intriguingly , rifampin and SJW , both activators of the human pregnane X receptor ( O75469 ) , induced CYP3A in both liver and intestine but induced P-gp only in intestine , whereas Q15149 , an activator of murine O75469 , did not induce P-gp in any tissue . DB01045 disposition was evaluated , and hepatic exposure to rifampin was comparable to intestine ; in contrast , brain concentrations were low . Overall , these observations demonstrate that P-gp induction in vivo is tissue-specific ; furthermore , there is a disconnect between P-gp induction and CYP3A induction that is tissue- and inducer-dependent , suggesting that O75469 activation alone is insufficient for P-gp induction in vivo . Tissue-specific factors and inducer pharmacokinetic/pharmacodynamic properties may underlie these observations . PLA(2) signaling is involved in calpain-mediated degradation of synaptic dihydropyrimidinase-like 3 protein in response to DB01221 excitotoxicity . Q14117 -like 3 ( Q14195 ) is believed to play a role in neuronal differentiation , axonal outgrowth and neuronal regeneration , as well as cytoskeleton organization . Recently we have shown that glutamate excitotoxicity and oxidative stress result in calpain-dependent cleavage of Q14195 , and that NOS plays a role in this process [ R . Kowara , Q . Chen , M. Milliken , B . Chakravarthy , Calpain-mediated truncation of dihydropyrimidinase-like 3 protein ( Q14195 ) in response to DB01221 and H2O2 toxicity , J. Neurochem. 95 ( 2005 ) 466-474 ; R. Kowara , K.L. Moraleja , B. Chakravarthy , Involvement of nitric oxide synthase and ROS-mediated activation of L-type voltage-gated Ca(2+) channels in DB01221 -induced Q14195 degradation , Brain Res . 1119 ( 2006 ) 40-49 ] . The present study investigates the involvement of PLA(2) signaling in DB01221 -induced Q14195 degradation . Exposure of rat primary cortical neurons ( Q15149 ) to PLA(2) and P35354 inhibitors significantly prevented DB01221 -induced Q14195 degradation . Since the metabolic product of PLA(2) signaling , PGE(2) , which augments toxic effect of DB01221 , is known to stimulate DB02527 , the effect of adenyl cyclase activator ( forskolin plus DB07954 ) and inhibitor ( MDL12,300 ) on DB01221 -induced Q14195 degradation was tested . Our data indicate that the activation of adenyl cyclase contributes to DB01221 -induced Q14195 degradation . Furthermore , DB02527 -dependent protein kinase ( PKA ) inhibitor PKI ( 14-22 ) provided additional evidence of PKA involvement in DB01221 -induced Q14195 degradation . In summary , the obtained data show the contribution of PLA(2) signaling to DB01221 -induced calpain activation and subsequent degradation of synaptic protein Q14195 . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Effects of DB01045 , a potent inducer of drug-metabolizing enzymes and an inhibitor of Q9Y6L6 /3 transport , on the single dose pharmacokinetics of anacetrapib . Anacetrapib is a novel cholesteryl ester transfer protein ( P11597 ) inhibitor in development for treatment of dyslipidemia . This open-label , fixed-sequence , 3-period study was intended to evaluate the potential of anacetrapib to be a victim of Q9Y6L6 /3 inhibition and strong CYP3A induction using acute and chronic dosing of rifampin , respectively , as a probe . In this study , 16 healthy subjects received 100 mg anacetrapib administered without rifampin ( Day 1 , Period 1 ) , with single-dose ( SD ) 600 mg rifampin ( Day 1 , Period 2 ) , and with multiple-dose ( MD ) 600 mg rifampin for 20 days ( Day 14 , Period 3 ) . Log-transformed anacetrapib AUC0-∞ and Cmax were analyzed by a linear mixed effects model . The GMRs and 90 % CIs for anacetrapib AUC0-∞ and Cmax were 1.25 ( 1.04 , 1.51 ) and 1.43 ( 1.13 , 1.82 ) for SD rifampin ( Period 2/Period 1 ) and 0.35 ( 0.29 , 0.42 ) and 0.26 ( 0.21 , 0.32 ) for MD rifampin ( Period 3/Period 1 ) , respectively . Anacetrapib was generally well tolerated in both the absence/presence of SD and MD rifampin . In conclusion , treatment with SD rifampin , which inhibits the Q9Y6L6 /3 transporter system , did not substantially influence the SD pharmacokinetics of anacetrapib , while chronic ( 20 days ) administration of rifampin , which strongly induces CYP3A isozymes , reduced mean systemic exposure to SD anacetrapib by 65 % . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB09026 : An orally active renin inhibitor . P00797 inhibitors are antihypertensive drugs that block the first step in the renin-angiotensin system . Their mechanism of action differs from that of the angiotensin-converting enzyme inhibitors and angiotensin-receptor antagonists , but like these drugs , renin inhibitors interrupt the negative feedback effects of angiotensin II on renin secretion . The renin-angiotensin-aldosterone system ( RAAS ) has long been recognized to play a significant role in hypertension pathophysiology . Certain agents that modify the RAAS can control blood pressure and improve cardiovascular outcomes . Optimization of this compound by Novartis led to the development of aliskiren - the only direct renin inhibitor which is clinically used as an antihypertensive drug . DB09026 is the first of a new class of antihypertensive agents . DB09026 is a new renin inhibitor of a novel structural class that has recently been shown to be efficacious in hypertensive patients after once-daily oral dosing . In short-term studies , it was effective in lowering blood pressure either alone or in combination with valsartan and hydrochlorothiazide , and had a low incidence of serious adverse effects . It was approved by the Food and Drug Administration in 2007 for the use as a monotherapy or in combination with other antihypertensives . Greater reductions in blood pressure have been achieved when aliskiren was used in combination with hydrochlorothiazide or an angiotensin-receptor blocker . The most common adverse effects reported in clinical trials were headache , fatigue , dizziness , diarrhea , and nasopharyngitis . DB09026 has not been studied in patients with moderate renal dysfunction ; as an RAAS-acting drug , it should be prescribed for such patients only with caution . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Identification of antithrombin-modulating genes . Role of O95461 , a gene encoding a bifunctional glycosyltransferase , in the secretion of proteins ? The haemostatic relevance of antithrombin together with the low genetic variability of P01008 , and the high heritability of plasma levels encourage the search for modulating genes . We used a hypothesis-free approach to identify these genes , evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study ( 352 individuals from 21 Spanish families ) . Despite no SNP reaching the genome wide significance threshold , we verified milder positive associations in 307 blood donors from a different cohort . This validation study suggested O95461 , a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides , as a potential modulator of antithrombin based on the significant association of one SNPs , rs762057 , with anti-FXa activity , particularly after adjustment for age , sex and P01008 rs2227589 genotype , all factors influencing antithrombin levels ( p = 0.02 ) . Additional results sustained this association . O95461 silencing inHepG2 and P29320 -EBNA cells did not affect P01008 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention . Milder effects were observed on α1-antitrypsin , prothrombin and transferrin . Our study suggests O95461 as the first known modifier of plasma antithrombin , and proposes a new role for O95461 in modulating extracellular secretion of certain glycoproteins . Effect of progesterone on intracellular Ca2+ homeostasis in human myometrial smooth muscle cells . Although it is well known that progesterone alters uterine contractility and plays an important role in maintenance of pregnancy , the biochemical mechanisms by which progesterone alters uterine contractility in human gestation are less clear . In this investigation we sought to identify progesterone-induced adaptations in human myometrial smooth muscle cells that may alter Ca2+ signaling in response to contractile agents . Cells were treated with vehicle or the progesterone analog medroxyprogesterone acetate ( DB00603 ) for 5 days , and intracellular free Ca2+ concentration ( [Ca2+]i ) was quantified after treatment with oxytocin ( OX ) or endothelin ( ET ) -1 . OX- and ET-1-induced increases in [Ca2+]i were significantly attenuated in cells pretreated with DB00603 in a dose-dependent manner . P06401 antagonists prevented the attenuated Ca2+ transients induced by DB00603 . P25101 and ETB receptor subtypes were expressed in myometrial cells , and treatment with DB00603 resulted in significant downregulation of P25101 and ETB receptor binding . DB00603 did not alter ionomycin-stimulated increases in [Ca2+]i and had no effect on inositol trisphosphate-dependent or -independent release of Ca2+ from internal Ca2+ stores . We conclude that adaptations of Ca2+ homeostasis in myometrial cells during pregnancy may include progesterone-induced modification of receptor-mediated increases in [Ca2+]i . Comparison of the efficacy of peripheral blood stem cell mobilization using G- P04141 alone from healthy donors and patients with hematologic malignancies . Peripheral blood stem cell ( PBSC ) collection using granulocyte colony-stimulating factor ( DB00099 ) alone is superior to the combination of chemotherapy and G- P04141 in terms of low morbidity , short duration of mobilization and low cost . We retrospectively compared the results of PBSC collection using G- P04141 alone in 11 patients with malignant lymphoma ( ML ) , 23 patients with plasma cell neoplasms ( Q15149 ) and 48 healthy donors . The geometric mean number of P28906 (+) cells/kg obtained on the first day of collection was 0.99 × 10(6)/kg in ML patients , 2.26 × 10(6)/kg in Q15149 patients , and 3.36 × 10(6)/kg in healthy donors . The probability of collecting at least 1 × 10(6)/kg P28906 (+) cells/kg during a single course of apheresis was 90.9 % in ML patients , 95.7 % in Q15149 patients , and 100 % in healthy donors . In a multiple regression analysis of the P28906 (+) cell yields on the first day of apheresis , we identified disease , the baseline white blood cell count ( WBC ) , platelet count , and lactate dehydrogenase as independent significant variables . Particularly , disease was strongly associated with the P28906 (+) cell yield , probably due to the difference in the number of previous chemotherapy cycles . In conclusion , the minimal dose of P28906 (+) cells for autologous transplantation was collected in almost all patients with hematological malignancies . However , patients who have received repeated cycles of chemotherapy , such as patients with ML , and those who have low WBC counts and/or platelet counts may be at higher risk for poor mobilization . CaMKII autonomy is substrate-dependent and further stimulated by Ca2+/calmodulin . A hallmark feature of Ca(2+)/calmodulin ( P62158 ) -dependent protein kinase II ( CaMKII ) regulation is the generation of Ca(2+)-independent autonomous activity by DB00156 -286 autophosphorylation . CaMKII autonomy has been regarded a form of molecular memory and is indeed important in neuronal plasticity and learning/memory . DB00156 -286-phosphorylated CaMKII is thought to be essentially fully active ( approximately 70-100 % ) , implicating that it is no longer regulated and that its dramatically increased Ca(2+)/ P62158 affinity is of minor functional importance . However , this study shows that autonomy greater than 15-25 % was the exception , not the rule , and required a special mechanism ( T-site binding ; by the T-substrates AC2 or Q13224 ) . Autonomous activity toward regular R-substrates ( including tyrosine hydroxylase and GluR1 ) was significantly further stimulated by Ca(2+)/ P62158 , both in vitro and within cells . Altered K(m) and V(max) made autonomy also substrate- ( and DB00171 ) concentration-dependent , but only over a narrow range , with remarkable stability at physiological concentrations . Such regulation still allows molecular memory of previous Ca(2+) signals , but prevents complete uncoupling from subsequent cellular stimulation . DB01045 Does not Significantly Affect the Expression of Small Heterodimer Partner in Primary Human Hepatocytes . The small/short heterodimer partner ( Q15466 , Q15466 ) is a nuclear receptor corepressor lacking a DNA binding domain . Q15466 is induced by bile acid-activated farnesoid X receptor ( Q96RI1 ) resulting in P22680 gene suppression . In contrast , O75469 ( O75469 ) activation by its ligands was recently suggested to inhibit Q15466 gene transactivation to maximize the induction of O75469 target genes . However , there are also conflicting reports in literature whether O75469 or rodent Pxr activation down-regulates Q15466 /Shp expression . Moreover , the O75469 -mediated regulation of the Q15466 gene has been studied only at the Q15466 mRNA and transactivation ( gene reporter assay ) levels . In this study , we studied the effect of rifampicin , a prototype O75469 ligand , on Q15466 mRNA , and protein expression in three primary human hepatocyte cultures . We found that Q15466 mRNA is not systematically down-regulated in hepatocyte in culture after 24 h treatment with rifampicin . Consistently , we did not observe down-regulation of Q15466 protein in primary human hepatocytes after 24 and 48 h of incubation with rifampicin . We can conclude that although we observed slight down-regulation of Q15466 mRNA and protein in several hepatocyte preparations , the phenomenon is unlikely critical for O75469 -mediated induction of its target genes . Hyperglycemia accelerates DB00171 -binding cassette transporter A1 degradation via an P29323 -dependent pathway in macrophages . An elevation in blood glucose concentration leads to increased risk of developing diabetes-associated atherosclerotic cardiovascular disease due to an excessive accumulation of cholesterol in arterial macrophages . DB00171 -binding cassette transporter A1 ( O95477 ) is an atheroprotective protein that mediates the export of cholesterol from macrophages . The present study aims to investigate the effect of hyperglycemia on the regulation of O95477 expression and to explore its underlying mechanisms of regulation in macrophages . Our results show that high glucose activates the extracellular signal-regulated kinases ( P29323 ) signaling pathway via reactive oxygen species ( ROS ) production , which in turn down-regulates O95477 mRNA and protein expression . This down-regulation is mediated by accelerating O95477 mRNA and protein degradation in macrophages exposed to high concentrations of glucose . Our results provide evidence for the first time that hyperglycemia inhibits O95477 expression by P29323 -modulated O95477 mRNA and protein stability . Overall , these results provide a mechanism for hyperglycemia-induced reduction in O95477 expression , which suggests a promising strategy for the treatment of diabetes-associated atherosclerosis . Q15149 regulates the signaling and trafficking of the HIV-1 co-receptor P61073 and plays a role in HIV-1 infection . The CXC chemokine P48061 and its cognate receptor P61073 play an important role in inflammation , human immunodeficiency virus ( HIV ) infection and cancer metastasis . The signal transduction and intracellular trafficking of P61073 are involved in these functions , but the underlying mechanisms remain incompletely understood . In the present study , we demonstrated that the P61073 formed a complex with the cytolinker protein plectin in a ligand-dependent manner in HEK293 cells stably expressing P61073 . The glutathione-S-transferase ( Q86UG4 ) - P61073 C-terminal fusion proteins co-precipitated with the full-length and the N-terminal fragments of plectin isoform 1 but not with the N-terminal deletion mutants of plectin isoform 1 , thereby suggesting an interaction between the N-terminus of plectin and the C-terminus of P61073 . This interaction was confirmed by confocal microscopic reconstructions showing co-distribution of these two proteins in the internal vesicles after ligand-induced internalization of P61073 in HEK293 cells stably expressing P61073 . Knockdown of plectin with RNA interference ( RNAi ) significantly inhibited ligand-dependent P61073 internalization and attenuated P61073 -mediated intracellular calcium mobilization and activation of extracellular signal regulated kinase 1/2 ( P27361 /2 ) . P48061 -induced chemotaxis of HEK293 cells stably expressing P61073 and of Jurkat T cells was inhibited by the plectin RNAi . Moreover , P61073 tropic HIV-1 infection in MAGI ( HeLa- P01730 -LTR-Gal ) cells was inhibited by the RNAi of plectin . Thus , plectin appears to interact with P61073 and plays an important role in P61073 signaling and trafficking and HIV-1 infection . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . [ Antibacterial activity of 16 antibiotics against Helicobacter pylori ] . The susceptibilities of 24 Helicobacter pylori isolates , which were originated from clinical materials , to 5 beta-lactam antibiotics [ benzylpenicillin ( DB01053 ) , ampicillin ( DB00415 ) , cephalothin ( DB00456 ) , ceftazidime ( DB00438 ) , cefotiam ( DB00229 ) and imipenem ( IPM ) ] , two macrolides [ clarithromycin ( P62158 ) and rokitamycin ( RKM ) ] , two aminoglycosides [ amikacin ( AMK ) and gentamicin ( GM ) ] , two new quinolones [ ciprofloxacin ( CPFX ) and levofloxacin ( LVFX ) ] , two tetracycline [ tetracycline ( TC ) and minocycline ( MINO ) ] , rifampicin ( Q9HBH0 ) and chloramphenicol ( CP ) were tested . All of the isolates showed similar susceptibilities against beta-lactam antibiotics . However , MICs of DB00229 and DB00438 were two- to four-fold higher than those of DB01053 , DB00415 , DB00456 and IPM , MICs of rokitamycin for the tested strains were higher than those of clarithromycin . MICs of CPFX and LVFX showed two-modal distributions . The first peak of distributions was observed between 0.06 to 0.5 microgram/ml and second one was between 4 to 16 micrograms/ml . These distributions suggested that MIC values of 4 to 16 micrograms/ml could result from the expression of a resistance mechanism . In addition , some of H. pylori strains were observed drug resistances between CP and AMK , new quinolones and AMK respectively . From the molecular epidemiological study , cryptic plasmids were detected from the 3 isolates among 24 strains tested .

MH_train_1068

MH_train_1068

MH_train_1068

interacts_with DB00159?

multiple_choice

Effect of endothelin receptor antagonist DB00559 on chronic hypoxia-induced inflammation and chemoafferent neuron adaptation in rat carotid body . Chronic hypoxia ( CH ) induces an inflammatory response in rat carotid body that is characterized by immune cell invasion and the expression of pro-inflammatory cytokines . In the present study , we have investigated the role of type-A endothelin ( P25101 ) receptors in the development of CH-induced inflammation . After 7 days of CH ( 380 Torr ) , double-label immunofluorescence studies demonstrated elevated levels of P25101 receptor and tyrosine hydroxylase ( TH ) in O(2)-sensitive type I cells . Following CH , P25101 receptors were also expressed on resident and invasive P08575 + immune cells distributed in tissue surrounding chemosensory cell lobules . Immnofluorescence and quantitative PCR studies showed that concurrent treatment with the P25101 /B receptor antagonist , DB00559 ( 200 mg/kg/day ) , blocked CH-induced ED-1+ macrophage invasion and the upregulation of cytokines , including interleukin-1β ( IL-1β ) , interleukin-6 ( P05231 ) , tumor necrosis factor α ( TNFα ) , and monocyte chemoattractant protein-1 ( P13500 ) . Moreover , DB00559 treatment blocked the CH-induced increases in expression of acid-sensitive ion channels ( ASICs ) in chemoafferent neurons in the petrosal ganglion ( PG ) . Our findings are consistent with the hypothesis that CH-induced inflammation involves the upregulation and release of ET-1 from type I cells . ET-1 may act in an autocrine/paracrine mechanism via P25101 receptors on chemosensory type I cells and immune cells to promote an inflammatory response . Glial response to lipopolysaccharide : possible role of endothelins . Glial inflammation plays a major role in the development of neurodegenerative diseases . Although endothelins ( ETs ) are known as modulators of inflammation in the periphery , little is known about their possible role in brain inflammation . Previously , we demonstrated that all three endothelins ( ET-1 , P20800 and P14138 ) enhanced unstimulated synthesis of the glial pro-inflammatory mediators , prostaglandin E₂ ( PGE₂ ) and nitric oxide ( NO ) . In the present study , glial cells were stimulated in an in vitro model of inflammation by incubation with the bacterial endotoxin lipopolysaccharide ( LPS ) . Indeed , the present study shows that ETs regulate basal and LPS-induced glial inflammation in an opposite fashion . Here we demonstrate that ETs significantly inhibited the LPS-induced glial synthesis of PGE₂ and NO , and each of the selective antagonists for P25101 and ETB receptors ( BQ123 and BQ788 respectively ) , significantly inhibited the ETs effects in LPS-treated cells . Similar results were observed when expression of key enzymes namely , cyclooxygenase-2 ( P35354 ) and inducible nitric oxide synthase ( P35228 ) in PG and NO synthesis respectively , was measured . ET-1 significantly enhanced the expression of both P35354 and P35228 . Whereas , it inhibited the LPS-induced expression of both enzymes . These observations suggest a novel neuro-immune feedback pathway through which inflammatory mediators ' synthesis is initially enhanced by ETs and are eventually blocked by the same neuropeptide when excessive production of inflammatory mediators occurs following an inflammatory insult . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . ω-3 fatty acid eicosapentaenoic acid attenuates P25189 +-induced neurodegeneration in fully differentiated human SH-SY5Y and primary mesencephalic cells . DB00159 ( EPA ) , a neuroactive omega-3 fatty acid , has been demonstrated to exert neuroprotective effects in experimental models of Parkinson 's disease ( PD ) , but the cellular mechanisms of protection are unknown . Here , we studied the effects of EPA in fully differentiated human SH-SY5Y cells and primary mesencephalic neurons treated with P25189 (+) . In both in-vitro models of PD , EPA attenuated an P25189 (+) -induced reduction in cell viability . EPA also prevented the presence of electron-dense cytoplasmic inclusions in SH-SY5Y cells . Then , possible mechanisms of the neuroprotection were studied . In primary neurons , EPA attenuated an P25189 (+) -induced increase in Tyrosine-related kinase B ( TrkB ) receptors . In SH-SY5Y cells , EPA down-regulated reactive oxygen species and nitric oxide . This antioxidant effect of EPA may have been mediated by its inhibition of neuronal NADPH oxidase and cyclo-oxygenase-2 ( P35354 ) , as P25189 (+) increased the expression of these enzymes . Furthermore , EPA prevented an increase in cytosolic phospholipase A2 ( P47712 ) , an enzyme linked with P35354 in the potentially pro-inflammatory arachidonic acid cascade . Lastly , EPA attenuated an increase in the bax:bcl-2 ratio , and cytochrome c release . However , EPA did not prevent mitochondrial enlargement or a decrease in mitochondrial membrane potential . This study demonstrated cellular mechanisms by which EPA provided neuroprotective effects in experimental PD . DB00159 modulates DB00091 -induced proinflammatory cytokine over-expression in osteoblastic cells in vitro . Several adverse outcomes are reported in subjects undergoing long term DB00091 ( DB00091 ) treatment . Severe osteopenia has been described in clinical and experimental reports , while beneficial effects of n-3 polyunsaturated fatty acids ( PUFAs ) on bone metabolism are recognized . In the present study we investigated the effects of n-3 versus n-6 PUFAs on osteoblastic cells treated with DB00091 , evaluating the expression of interleukin ( IL ) -1 & #223 ; , interleukin-6 ( P05231 ) , inducible nitric oxide synthase ( P35228 ) , and cyclooxygenase-2 ( P35354 ) in two different experimental protocols and the production of P05231 , IL-1 & #223 ; , and tumor necrosis factor alpha ( TNFalpha ) in cells challenged simultaneously with DB00091 and eicosapentaenoic acid ( EPA ) for 48h . IL-1 & #223 ; and P05231 up-regulation , induced by DB00091 , was counteracted by the addition of EPA in both protocols ; on the contrary , arachidonic acid ( AA ) magnified DB00091 the effects . P35354 and P35228 levels were not modified by DB00091 treatment . These in vitro results , that substantiate clinical reports of DB00091 -induced osteopenia , demonstrate a beneficial effect of EPA on DB00091 -altered cytokine profile , opening new perspectives in the non-pharmacological management of adverse outcomes in DB00091 -treated patients . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Biological activity of (lipo)polysaccharides of the exopolysaccharide-deficient mutant Rt120 derived from Rhizobium leguminosarum bv. trifolii strain Q96RJ0 . Lipopolysaccharides ( LPS ) from Rhizobium leguminosarum biovar trifolii Q96RJ0 ( RtTA1 ) and its mutant Rt120 in the pssBpssA intergenic region as well as degraded polysaccharides ( DPS ) derived from the LPS were elucidated in terms of their chemical composition and biological activities . The polysaccharide portions were examined by methylation analysis , MALDI-TOF mass spectrometry , and (1)H NMR spectroscopy . A high molecular mass carbohydrate fraction obtained from Rt120 DPS by Sephadex G-50 gel chromatography was composed mainly of L-rhamnose , 6-deoxy-L-talose , D-galactose , and D-galacturonic acid , whereas that from RtTA1 DPS contained L-fucose , 2-acetamido-2,6-dideoxy-D-glucose , D-galacturonic acid , 3-deoxy-3-methylaminofucose , D-glucose , D-glucuronic acid , and heptose . Relative intensities of the major (1)H NMR signals for O-acetyl and N-acetyl groups were 1 : 0.8 and 1 : 1.24 in DPS of Rt120 and RtTA1 , respectively . The intact mutant LPS exhibited a twice higher lethal toxicity than the wild type LPS . A higher in vivo production of TNFα and P05231 after induction of mice with Rt120 LPS correlated with the toxicity , although the mutant LPS induced the secretion of IL-1β and IFNγ more weakly than RtTA1 LPS . A polysaccharide obtained by gel chromatography on Bio-Gel P-4 of the high molecular mass material from Rt120 had a toxic effect on tumor HeLa cells but was inactive against the normal human skin fibroblast cell line . The polysaccharide from RtTA1 was inactive against either cell line . The potent inhibitory effect of the mutant DPS on tumor HeLa cells seems to be related with the differences in sugar composition . Influence of cystic fibrosis transmembrane conductance regulator on gene expression in response to Pseudomonas aeruginosa infection of human bronchial epithelial cells . Chronic lung infection by Pseudomonas aeruginosa causes significant morbidity in cystic fibrosis patients initiated by the failure of innate immune responses . We used microarray analysis and real-time PCR to detect transcriptional changes associated with cytokine production in isogenic bronchial epithelial cell lines with either wild-type ( WT ) or mutant cystic fibrosis transmembrane conductance regulator ( P13569 ) in response to P. aeruginosa infection . The transcription of four NF-kappaB-regulated cytokine genes was maximal in the presence of WT P13569 : the interleukin-8 ( P10145 ) , P05231 , P09341 , and intracellular adhesion molecule 1 ( P05362 ) genes . Analysis of protein expression in two cell lines paired for wild-type and mutant P13569 with three P. aeruginosa strains showed P05231 and P10145 expressions were consistently enhanced by the presence of WT P13569 in both cell lines with all three strains of P. aeruginosa , although some strains gave small P10145 increases in cells with mutant P13569 . P09341 production showed consistent enhancement in cells with WT P13569 using all three bacterial strains in one cell line , whereas in the other cell line , P09341 showed a significant increase in cells with either WT or mutant P13569 . P05362 was unchanged at the protein level in one of the cell lines but did show mild enhancement with WT P13569 in the other cell pair . Inhibitions of NF-kappaB prior to infection indicated differing degrees of dependence on NF-kappaB for production of the cytokines , contingent on the cell line . Cytokine effectors of innate immunity to P. aeruginosa were found to be positively influenced by the presence of WT P13569 , indicating a role in resistance to P. aeruginosa infection . Di(2-ethylhexyl) phthalate induces apoptosis through peroxisome proliferators-activated receptor-gamma and P29323 1/2 activation in testis of Sprague-Dawley rats . Di(2-ethylhexyl) phthalate ( DEHP ) is a well-known hepatic and reproductive toxicant whose toxicity may be mediated by peroxisome proliferators-activated receptor ( Q07869 ) . This study examined the effects of DEHP on the expression of Q07869 -regulated genes involved in testicular cells apoptosis . Sprague-Dawley male rats were treated orally with 250 , 500 , or 750 mg/kg/d DEHP for 28 d , while control rats were given corn oil . The levels of cell cycle regulators ( P06400 , cyclins , CDKs , and P38936 ) and apoptosis-related proteins were analyzed by Western blot analysis . The role of P37231 ( P37231 ) , class B scavenger receptor type 1 ( SR-B1 ) , and P27361 /2 was further studied to examine the signaling pathway for DEHP-induced apoptosis . Results showed that the levels of pRB , cyclin D , P24941 , cyclin E , and P11802 were significantly lower in rats given 500 and 750 mg/kg/d DEHP , while levels of P38936 were significantly higher in rat testes . Dose-dependent increases in P37231 and RXRalpha proteins were observed in testes after DEHP exposure , while there was a significant decrease in RXRgamma protein levels . In addition to P37231 , DEHP also significantly increased SR-B1 mRNA and phosphorylated P27361 /2 protein levels . Furthermore , DEHP treatment induced pro-caspase-3 and cleavage of its substrate protein , poly(ADP-ribose) polymerase ( PARP ) , in a dose-dependent manner . Data suggest that DEHP exposure may induce the expression of apoptosis-related genes in testes through induction of P37231 and activation of the P27361 /2 pathway . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . Thiazolidinediones : effects on insulin resistance and the cardiovascular system . Thiazolidinediones ( TZDs ) have been used for the treatment of hyperglycaemia in type 2 diabetes for the past 10 years . They may delay the development of type 2 diabetes in individuals at high risk of developing the condition , and have been shown to have potentially beneficial effects on cardiovascular risk factors . TZDs act as agonists of peroxisome proliferator-activated receptor-gamma ( P37231 ) primarily in adipose tissue . P37231 receptor activation by TZDs improves insulin sensitivity by promoting fatty acid uptake into adipose tissue , increasing production of adiponectin and reducing levels of inflammatory mediators such as tumour necrosis factor-alpha ( P01375 ) , plasminogen activator inhibitor-1( P05121 ) and interleukin-6 ( P05231 ) . Clinically , TZDs have been shown to reduce measures of atherosclerosis such as carotid intima-media thickness ( CIMT ) . However , in spite of beneficial effects on markers of cardiovascular risk , TZDs have not been definitively shown to reduce cardiovascular events in patients , and the safety of rosiglitazone in this respect has recently been called into question . Dual Q07869 /gamma agonists may offer superior treatment of insulin resistance and cardioprotection , but their safety has not yet been assured . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Influence of labor on neonatal neutrophil apoptosis , and inflammatory activity . Neutrophil apoptosis is impaired in neonates , and this contributes to prolonged inflammation and tissue injury in infants after infection or trauma . In the present studies , we investigated whether labor generates mediators that further suppress apoptosis . We found that neutrophil apoptosis was reduced in neonates exposed to labor , when compared with infants delivered by cesarean section before labor . This was not due to alterations in caspase-3 or inhibitor of apoptosis protein-2 ( IAP-2 ) . In contrast , labor primed neutrophils to express tumor necrosis factor alpha ( P01375 ) , suggesting that proinflammatory mediators contribute to reduced apoptosis after labor . Eicosanoids generated via cyclooxygenase-2 ( Cox-2 ) and lipoxygenase ( Lox ) also regulate neutrophil apoptosis . 15-Lox , which generates proapoptotic lipoxins , but not Cox-2 , was greater in neutrophils before labor , relative to cells exposed to labor . Anti-inflammatory eicosanoids exert their effects in part via peroxisome proliferator-activated receptor gamma ( P37231 ) . Expression of gelatinase-associated lipocalin and catalase , two markers of P37231 activity , were increased in neonatal neutrophils before labor , relative to cells exposed to labor . These findings suggest that the anti-inflammatory environment is maintained before labor , in part , by eicosanoids . Although increased neutrophil longevity after labor is important for host defense in the immediate newborn period , it may contribute to inflammatory or oxidative injury in susceptible infants . Targeting other abnormal signaling pathways in sarcoma : P00533 in synovial sarcomas , P37231 in liposarcomas . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response . To investigate the mechanism of action by which a new anti-inflammatory active compound , 1-O-acetylbritannilactone ( P00519 ) isolated from Inula britannica-F. , inhibits inflammatory responses in vascular smooth muscle cells ( VSMCs ) . Enzyme immunoassay was used to measure the levels of prostandin E(2) ( PGE(2) ) production . Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB ( NF-kappaB ) p65 and the expression of IkappaB-alpha , pIkappaB-alpha and cyclooxygenase-2 ( P35354 ) . Electrophoretic mobility shift assays ( EMSA ) were used to detect DNA-binding activity of NF-kappaB in VSMCs . P00519 ( 5 , 10 , 20 micrommol/l ) had several concentration-dependent effects , including inhibition of lipopolysaccharide ( LPS ) -induced PGE(2) production and P35354 expression , and blockade of NF-kappaB activation and translocation . These effects were owing to reductions in IkappaB-alpha phosphorylation and degradation induced by LPS . In addition , P00519 directly inhibited the binding of active NF-kappaB to specific DNA cis-element . These results indicate that P00519 is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene P35354 expression . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . Omega-3 polyunsaturated fatty acids and inflammatory processes : nutrition or pharmacology ? DB00159 ( EPA ) and docosahexaenoic acid ( DB01708 ) are n-3 fatty acids found in oily fish and fish oil supplements . These fatty acids are able to inhibit partly a number of aspects of inflammation including leucocyte chemotaxis , adhesion molecule expression and leucocyte-endothelial adhesive interactions , production of eicosanoids like prostaglandins and leukotrienes from the n-6 fatty acid arachidonic acid , production of inflammatory cytokines and T cell reactivity . In parallel , EPA gives rise to eicosanoids that often have lower biological potency than those produced from arachidonioc acid and EPA and DB01708 give rise to anti-inflammatory and inflammation resolving resolvins and protectins . Mechanisms underlying the anti-inflammatory actions of n-3 fatty acids include altered cell membrane phospholipid fatty acid composition , disruption of lipid rafts , inhibition of activation of the pro-inflammatory transcription factor nuclear factor kappa B so reducing expression of inflammatory genes , activation of the anti-inflammatory transcription factor P37231 ( i.e. peroxisome proliferator activated receptor γ ) and binding to the G protein coupled receptor Q5NUL3 . These mechanisms are interlinked . In adult humans , an EPA plus DB01708 intake greater than 2 g day⁻¹ seems to be required to elicit anti-inflammatory actions , but few dose finding studies have been performed . Animal models demonstrate benefit from n-3 fatty acids in rheumatoid arthritis ( RA ) , inflammatory bowel disease ( Q9UKU7 ) and asthma . Clinical trials of fish oil in patients with RA demonstrate benefit supported by meta-analyses of the data . Clinical trails of fish oil in patients with Q9UKU7 and asthma are inconsistent with no overall clear evidence of efficacy . P05305 -induced arachidonic acid release by cytosolic phospholipase A2 activation in rat vascular smooth muscle via extracellular signal-regulated kinases pathway . The present study investigates whether endothelin-1 ( ET-1 ) , like noradrenaline ( NA ) , stimulates the release of arachidonic acid ( AA ) via cytosolic phospholipase A2 ( P47712 ) in rat tail artery . In tail artery segments labelled with [3H]AA , ET-1-induced AA release in a concentration-dependent manner with an EC50 of 1.3 nM . The effect of ET-1 was inhibited by DB00559 and was insensitive to BQ788 , suggesting the involvement of P25101 receptor . The stimulation of AA release induced by ET-1 was prevented by arachydonyl trifluoromethyl ketone ( AACOCF3 ) , a selective inhibitor of P47712 and not by RHC80267 , a diacylglycerol lipase inhibitor . Furthermore , PD98059 , inhibitor of mitogen-activated protein kinase kinase ( MEK ) cascade and calphostin C , a protein kinase C ( PKC ) inhibitor , prevented the stimulation of AA release induced by ET-1 and NA . Immunoblotting of the cytosolic fraction of rat tail arteries stimulated with ET-1 or NA showed an increase in extracellular signal-regulated kinases ( ERKs ) phosphorylation and this effect was abolished by calphostin C treatment . These findings show that in rat tail artery ET-1 and NA induce a sequential activation of protein kinase C and extracellular signal-regulated kinases that results in stimulation of AA release via P47712 activation . This may represent a general pathway by which G-proteins coupled receptors stimulate AA release and its metabolites in vascular smooth muscle . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . The impact of biological agents interfering with receptor/ligand binding in the immune system . We herein discuss the impact of biological agents based on the ability of monoclonal antibodies to target specific molecules . This approach has given to clinical immunologists a spectrum of drugs able to manipulate the immune system . In the first session , we discuss drugs targeting T-cell function by : ( 1 ) targeting P10747 mediated costimulation ( DB01281 and DB06681 ) ; ( 2 ) interfering with interleukin-2 receptor ( DB00074 and DB00111 ) ; ( 3 ) blocking cell adhesion and homing ( DB00092 , DB00095 , DB00108 ) . The second session is dedicated to drugs targeting cytokines or their receptors . The best known and largely experimented case is represented by drugs targeting tumor necrosis factor ( P01375 ) ( DB00065 , Adalilumab , Certolizumab ) or its p75 receptor ( DB00005 ) . However , newer products are now available to target other inflammatory cytokines including P05231 , P10145 , IL-12 , P40933 , Q14116 , IL-23 . These agents have the potential to become powerful tools in the control of several immune-mediated diseases , especially auto-immune and inflammatory ones . They traslate into reality the prediction that antibodies will eventually become " magic bullets which seek their own target " ( P. Ehrich , 1906 ) . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . JTE-522 , a selective P35354 inhibitor , inhibits cell proliferation and induces apoptosis in RL95-2 cells . AIM : To investigate whether JTE-522 [ 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide ] , a selective P35354 inhibitor , can induce apoptosis and inhibit cell proliferation in human endometrial cancer cell line RL95-2 cells and to explore the molecular mechanisms . METHODS : [ 3-(4,5)-dimethylthiazol-2-yl ] -2,5-diphenyl tetrazolium bromide ( MTT ) , DNA ladder , enzyme-linked immunosorbent assay ( ELISA ) , flow cytometry , RT-PCR , and Western blot analysis were employed to investigate effect of JTE-522 on human endometrial cancer cell line RL95-2 cells and the related molecular mechanisms . RESULTS : JTE-522 inhibited the growth of RL95-2 cells and induced the apoptosis . Furthermore , it arrested G0/ P55008 phase and inhibited S phase in RL95-2 cells . JTE-522 inhibited the expressions of P35354 mRNA , phosphorylated Rb , and P11802 proteins , while increased the levels of p53 , P38936 , cyclin D1 proteins , and the activity of caspase-3 in RL95-2 cells . CONCLUSION : JTE-522 inhibits cell proliferation and induces apoptosis in RL95-2 cells , which may be associated with the activation of caspase-3-like proteases , down-regulation of the expression of P35354 mRNA , phosphorylated Rb , and P11802 proteins , and up-regulation of the expressions of p53 , P38936 , and cyclin D1 proteins . Genetic mechanism of aspirin-induced urticaria/angioedema . PURPOSE OF REVIEW : DB00945 -induced urticaria/angioedema is a major aspirin-related hypersensitivity often associated with aspirin-intolerant asthma . Genetic studies on aspirin-intolerant asthma have shown chronic overproduction of cysteinyl leukotrienes . The genetic analysis of aspirin-induced urticaria/angioedema is limited , however . RECENT FINDINGS : A recent study on HLA genotypes has suggested that the HLA alleles DRB11302 and DQB10609 may be genetic markers for aspirin-induced urticaria/angioedema . A polymorphism study that examined nine single-nucleotide polymorphisms of five leukotriene-related genes [ P09917 ( encoding P09917 ) , P20292 ( P09917 -activating protein ) , P35354 ( cyclooxygenase 2 ) , Q16873 ( leukotriene C4 synthase ) , and Q9Y271 ( cysteinyl leukotriene receptor 1 ) ] found that promoter polymorphisms of P09917 ( -1708A > G ) and Q9Y271 ( -634C > T ) were significantly different between aspirin-intolerant asthma and aspirin-induced urticaria/angioedema , suggesting different contributions to the lipoxygenase pathway . A second polymorphism study , conducted on histamine-related genes , did not find any significant associations with aspirin-induced urticaria/angioedema for the genes P50135 ( encoding histamine N-methyltransferase ) , P35367 or P25021 ( encoding histamine receptor types 1 and 2 respectively ) , or the gene encoding high-affinity IgE receptor Ibeta ( FcepsilonRIbeta ) ; however , the FcepsilonRIalpha gene promoter polymorphism was significantly associated with aspirin-induced urticaria/angioedema . This finding has been supported by in vitro functional studies . SUMMARY : The HLA alleles DRB11302 and DQB10609 , and the P09917 and FcepsilonRIalpha promoter polymorphisms , may contribute to the pathogenesis of aspirin-induced urticaria/angioedema . Further investigation to identify candidate genetic markers would help to elucidate the pathogenic mechanism of this condition . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . Dynamic genetic linkage of intermediate blood pressure phenotypes during postural adaptations in a founder population . Blood pressure ( BP ) is a dynamic phenotype that varies rapidly to adjust to changing environmental conditions . Standing upright is a recent evolutionary trait , and genetic factors that influence postural adaptations may contribute to BP variability . We studied the effect of posture on the genetics of BP and intermediate BP phenotypes . We included 384 sib-pairs in 64 sib-ships from families ascertained by early-onset hypertension and dyslipidemia . Blood pressure , three hemodynamic and seven neuroendocrine intermediate BP phenotypes were measured with subjects lying supine and standing upright . The effect of posture on estimates of heritability and genetic covariance was investigated in full pedigrees . Linkage was conducted on 196 candidate genes by sib-pair analyses , and empirical estimates of significance were obtained . A permutation algorithm was implemented to study the postural effect on linkage . ADRA1A , APO , CAST , Q9Y5Q5 , P34998 , P24530 , P09038 , GC , P17302 , Q92953 , P08254 , P01303 , P08235 , P26678 , P37173 , P25445 , and P34981 showed evidence of linkage with any phenotype in the supine position and not upon standing , whereas P15121 , P16671 , P25101 , P12259 , P14780 , PKD2 , P27169 , P37231 , Q9UBK2 , P17252 , and P07949 were specifically linked to standing phenotypes . Genetic profiling was undertaken to show genetic interactions among intermediate BP phenotypes and genes specific to each posture . When investigators perform genetic studies exclusively on a single posture , important genetic components of BP are missed . Supine and standing BPs have distinct genetic signatures . Standardized maneuvers influence the results of genetic investigations into BP , thus reflecting its dynamic regulation . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . DB00159 inhibits prostaglandin D2 generation by inhibiting cyclo-oxygenase-2 in cultured human mast cells . BACKGROUND : DB00159 ( EPA ) is catalysed by cyclo-oxygenase ( P36551 ) , as is arachidonic acid , and is a competitive inhibitor of arachidonate metabolism . OBJECTIVES : We examined the effect of EPA on prostaglandin ( PG ) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation . METHODS : Cultured human mast cells were incubated with EPA ( 1 micromol/L ) for 20 h , then challenged with anti-IgE incubation after treatment with IgE . At the same time , P36551 inhibitors were tested to identify P23219 and P35354 activity . PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with P36551 inhibitors and EPA . DB11320 in the culture medium and in cells was assayed with the HPLC-fluorescent method . PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method . RESULTS : Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation , it did decrease PGD2 generation by inhibiting the P35354 pathway . In contrast , in the cell-free homogenate of cultured human mast cells , EPA inhibited both P23219 and P35354 activities . CONCLUSION : Pre-incubation with EPA primarily affects the P35354 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation . These findings suggest that P23219 and P35354 have different substrate flow systems in mast cells . They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells . P37231 controls Muc1 transcription in trophoblasts . The nuclear receptor peroxisome proliferator-activated receptor gamma ( PPARgamma ) is essential for placental development . Here , we show that the mucin gene Muc1 is a PPARgamma target , whose expression is lost in PPARgamma null placentas . During differentiation of trophoblast stem cells , PPARgamma is strongly induced , and Muc1 expression is upregulated by the PPARgamma agonist rosiglitazone . Muc1 promoter is activated strongly and specifically by liganded PPARgamma but not PPARalpha or PPARdelta . A Q07869 binding site ( DR1 ) in the proximal Muc1 promoter acts as a basal silencer in the absence of PPARgamma , and its cooperation with a composite upstream enhancer element is both necessary and sufficient for PPARgamma-dependent induction of Muc1 . In the placenta , P15941 protein is localized exclusively to the apical surface of the labyrinthine trophoblast around maternal blood sinuses , resembling its luminal localization on secretory epithelia . Last , variably penetrant maternal blood sinus dilation in Muc1-deficient placentas suggests that Muc1 regulation by PPARgamma contributes to normal placental development but also that the essential functions of PPARgamma in the organ are mediated by other targets . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Antiinflammatory effect of lactic acid bacteria : inhibition of cyclooxygenase-2 by suppressing nuclear factor-kappaB in Raw264.7 macrophage cells . Lactobacillus casei 3260 ( L. casei 3260 ) was evaluated in relation to the inflammatory response mediated by lipopolysaccharide ( LPS ) -induced nuclear factor-kappaB ( NF-kappaB ) and cyclooxygenase-2 ( P35354 ) expression in Raw264.7 macrophage cells . The treatment of Raw264.7 cells with L. casei 3260 significantly inhibited the secretion of tumor necrosis factor-alpha ( P01375 ) and prostaglandins E2 ( DB00917 ) , followed by suppression of P35354 . To clarify the molecular mechanism , the inhibitory effect of L. casei 3260 on the NF-kappaB signaling pathway was examined based on the luciferase reporter activity . Although the treatment of Raw264.7 cells with L. casei 3260 did not affect the transcriptional activity of NF-kappaB , it did inhibit NF-kappaB activation , as determined by the cytosolic p65 release and degradation of I-kappaBalpha . Therefore , these findings suggest that the suppression of P35354 through inhibiting the NF-kappaB activation by LPS may be associated with the antiinflammatory effects of L. casei 3260 on Raw264.7 cells . [ Comparative study of somatic oncogene mutations in normal thyroid tissues and thyroid neoplasms ] . It is established that numerous somatic oncogene mutation ( P15056 , P01111 , P01112 , P01116 ) and gene translocations ( P07949 /PTC , Q06710 / P37231 ) are associated with the development of thyroid cancer . In this study 22 intraoperative thyroid tissue samples ( 11 pathologic and 11 normal ) were examined . Somatic single nucleotide polymorphisms were analyzed by LigthCycler melting method , while translocations were identified by real-time polymerase chain reaction technique . In tumorous sample 3 P15056 , 2 P01111 and one P01112 mutations were found , as well as one P07949 / Q13635 translocation . Results confirm international data showing that these oncogene mutations and translocations are linked to thyroid cancer . Cytological examination completed with genetic data may support the diagnosis of thyroid malignancies . In addition , genetic alterations may indicate malignant transformation and may become prognostic factors in future . P37231 promotes epithelial to mesenchymal transformation by Rho GTPase-dependent activation of P27361 /2 . P37231 ( PPARgamma ) causes epithelial to mesenchymal transformation ( EMT ) in intestinal epithelial cells , as evidenced by reorganization of the actin cytoskeleton , acquisition of a polarized , mesenchymal cellular morphology , increased cellular motility , and colony scattering . This response is due to activation of Cdc42 , resulting in P38936 -activated kinase-dependent phosphorylation and activation of Q02750 DB00133 (298) and activation of P27361 /2 . Dominant negative Q02750 , P36507 , and P28482 block PPARgamma-induced EMT , whereas constitutively active Q02750 and P36507 induce a mesenchymal phenotype similar to that evoked by PPARgamma . PPARgamma also stimulates P27361 /2 phosphorylation in the intestinal epithelium in vivo . PPARgamma induces the P42336 subunit of phosphoinositide 3-kinase ( PI3K ) , and inhibition of PI3K blocks PPARgamma-dependent phosphorylation of Q02750 DB00133 (298) , activation of P27361 /2 , and EMT . We conclude that PPARgamma regulates the motility of intestinal epithelial cells through a mitogen-activated protein kinase cascade that involves PI3K , Cdc42 , P38936 -activated kinase , Q02750 , and P27361 /2 . Regulation of cellular motility through Rho family GTPases has not been previously reported for nuclear receptors , and elucidation of the mechanism that accounts for the role of PPARgamma in regulating motility of intestinal epithelial cells provides fundamental new insight into the function of this receptor during renewal of the intestinal epithelium .

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interacts_with DB00470?

multiple_choice

Cannabinoids and schizophrenia : therapeutic prospects . Approximately one third of patients diagnosed with schizophrenia do not achieve adequate symptom control with standard antipsychotic drugs ( APs ) . Some of these may prove responsive to clozapine , but non-response to APs remains an important clinical problem and cause of increased health care costs . In a significant proportion of patients , schizophrenia is associated with natural and iatrogenic metabolic abnormalities ( obesity , dyslipidaemia , impaired glucose tolerance or type 2 diabetes mellitus ) , hyperadrenalism and an exaggerated Q9Y251 response to stress , and chronic systemic inflammation . The endocannabinoid system ( ECS ) in the brain plays an important role in maintaining normal mental health . ECS modulates emotion , reward processing , sleep regulation , aversive memory extinction and Q9Y251 axis regulation . ECS overactivity contributes to visceral fat accumulation , insulin resistance and impaired energy expenditure . The cannabis plant synthesises a large number of pharmacologically active compounds unique to it known as phytocannabinoids . In contrast to the euphoric and pro-psychotic effects of DB00470 ( THC ) , certain non-intoxicating phytocannabinoids have emerged in pre-clinical and clinical models as potential APs . Since the likely mechanism of action does not rely upon dopamine D2 receptor antagonism , synergistic combinations with existing APs are plausible . The anti-inflammatory and immunomodulatory effects of the non-intoxicating phytocannabinoid cannabidiol ( DB09061 ) are well established and are summarised below . Preliminary data reviewed in this paper suggest that DB09061 in combination with a P21554 receptor neutral antagonist could not only augment the effects of standard APs but also target the metabolic , inflammatory and stress-related components of the schizophrenia phenotype . Eupolyphaga sinensis walker displays inhibition on hepatocellular carcinoma through regulating cell growth and metastasis signaling . Tumor growth and metastasis are responsible for most cancer patients ' deaths . Here , we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis . Compared with proliferation , colony formation , transwell assay and transplantable tumor in nude mouse in vitro and vivo , eupolyphaga sinensis walker extract ( ESWE ) showed good inhibition on the SMMC-7721 cell growth and metastasis . Using genome-wide microarray analysis , we found the down-regulated growth and metastasis factors , and selected down-regulated genes were confirmed by real-time PCR . Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE . Moreover , our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ , Akt , m-TOR , Erk1/2 , MEK-2 , Raf and JNK-1 , but also increasing cyclin D1 protein levels and decreasing amount of cyclin E , cyclin B1 and cdc2 of the cycle proteins . At the same time , ESWE reduced P08253 , P14780 and P61073 , P00747 , NFκB and P04637 activities . Overall , our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC , AKT , MAPK signaling and related metastasis signaling , having potential in cancer therapy . Arsenic decreases antinociceptive activity of paracetamol : possible involvement of serotonergic and endocannabinoid receptors . We assessed whether repeated arsenic exposure can decrease paracetamol-mediated antinociception by modulating serotonergic and endocannabinoid pathways . Rats were preexposed to elemental arsenic ( 4ppm ) as sodium arsenite through drinking water for 28 days . Next day paracetamol 's ( 400mg/kg , oral ) antinociceptive activity was assessed through formalin-induced nociception . Serotonin content and gene expression of P08908 , 5- Q13049 and P21554 receptors were evaluated in brainstem and frontal cortex . Arsenic decreased paracetamol-mediated analgesia . DB00316 , but not arsenic , increased serotonin content in these regions . Arsenic attenuated paracetamol-mediated increase in serotonin level . DB00316 did not alter P08908 expression , but caused down-regulation of 5- Q13049 and up-regulation of P21554 receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5- Q13049 was more pronounced . Arsenic did not modify paracetamol 's effect on P08908 expression , but reduced paracetamol-mediated down-regulation of 5- Q13049 and reversed up-regulation of P21554 receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5- Q13049 and antinociceptive P21554 receptors . Targeting CB2 cannabinoid receptors as a novel therapy to treat malignant lymphoblastic disease . In the current study , we examined whether ligation of CB2 receptors would lead to induction of apoptosis in tumors of immune origin and whether CB2 agonist could be used to treat such cancers . Exposure of murine tumors EL-4 , LSA , and P815 to DB00470 ( THC ) in vitro led to a significant reduction in cell viability and an increase in apoptosis . Exposure of EL-4 tumor cells to the synthetic cannabinoid HU-210 and the endogenous cannabinoid anandamide led to significant induction of apoptosis , whereas exposure to WIN55212 was not effective . Treatment of EL-4 tumor-bearing mice with THC in vivo led to a significant reduction in tumor load , increase in tumor-cell apoptosis , and increase in survival of tumor-bearing mice . Examination of a number of human leukemia and lymphoma cell lines , including Jurkat , Molt-4 , and Sup-T1 , revealed that they expressed CB2 receptors but not P21554 . These human tumor cells were also susceptible to apoptosis induced by THC , HU-210 , anandamide , and the CB2-selective agonist JWH-015 . This effect was mediated at least in part through the CB2 receptors because pretreatment with the CB2 antagonist SR144528 partially reversed the THC-induced apoptosis . Culture of primary acute lymphoblastic leukemia cells with THC in vitro reduced cell viability and induced apoptosis . Together , the current data demonstrate that CB2 cannabinoid receptors expressed on malignancies of the immune system may serve as potential targets for the induction of apoptosis . Also , because CB2 agonists lack psychotropic effects , they may serve as novel anticancer agents to selectively target and kill tumors of immune origin . P04818 genotype-directed chemotherapy for patients with gastric and gastroesophageal junction cancers . BACKGROUND : Retrospective studies indicate associations between TSER ( thymidylate synthase enhancer region ) genotypes and clinical outcomes in patients receiving DB00544 based chemotherapy , but well-controlled prospective validation has been lacking . METHODS : In this phase II study ( NCT00515216 registered through ClinicalTrials.gov , http://clinicaltrials.gov/show/NCT00515216 ) , patients with " good risk " TSER genotypes ( at least one TSER*2 allele ) were treated with FOLFOX chemotherapy to determine whether prospective patient selection can improve overall response rates ( ORR ) in patients with gastric and gastroesophageal junction ( GEJ ) cancers , compared with historical outcomes in unselected patients ( estimated 43 % ) . RESULTS : The ORR in genotype-selected patients was Q04695 % ( 9 partial responses out of 23 evaluable patients , 95 % CI , 22.2 to 59.2 ) , not achieving the primary objective of improving ORR . An encouraging disease control rate ( DCR , consisting of partial responses and stable diseases ) of 95.7 % was noted and patients with homozygous TSER*2 genotype showed better tumor response . CONCLUSIONS : In this first prospective , multi-institutional study in patients with gastric or GEJ cancers , selecting patients with at least one TSER*2 allele did not improve the ORR but led to an encouraging DCR . Further studies are needed to investigate the utility of selecting patients homozygous for the TSER*2 allele and additional genomic markers in improving clinical outcomes for patients with gastric and GEJ cancers . TRIAL REGISTRATION : ClinicalTrials.gov NCT00515216 . Distribution of cannabinoid receptors in the central and peripheral nervous system . P21554 cannabinoid receptors appear to mediate most , if not all of the psychoactive effects of DB00470 and related compounds . This G protein-coupled receptor has a characteristic distribution in the nervous system : It is particularly enriched in cortex , hippocampus , amygdala , basal ganglia outflow tracts , and cerebellum -- a distribution that corresponds to the most prominent behavioral effects of cannabis . In addition , this distribution helps to predict neurological and psychological maladies for which manipulation of the endocannabinoid system might be beneficial . P21554 receptors are primarily expressed on neurons , where most of the receptors are found on axons and synaptic terminals , emphasizing the important role of this receptor in modulating neurotransmission at specific synapses . While our knowledge of P21554 localization in the nervous system has advanced tremendously over the past 15 years , there is still more to learn . Particularly pressing is the need for ( 1 ) detailed anatomical studies of brain regions important in the therapeutic actions of drugs that modify the endocannabinoid system and ( 2 ) the determination of the localization of the enzymes that synthesize , degrade , and transport the endocannabinoids . ∆(9)- DB00470 decreases O60936 receptor density and mRNA levels in human SH-SY5Y cells . Several studies demonstrated a cross-talk between the opioid and cannabinoid system . The O60936 receptor and its endogenous ligand nociceptin/orphanin FQ represent an opioid-related functional entity that mediates some non-classical opioid effects . The relationship between cannabinoid and nociceptin/ O60936 system is yet poorly explored . In this study , we used the neuroblastoma SH-SY5Y cell line to investigate the effect of DB00470 ( ∆(9)-THC ) on nociceptin/ O60936 system . Results revealed that the exposure to ∆(9)-THC ( 100 , 150 , and 200 nM ) for 24 h produces a dose-dependent O60936 receptor B ( max ) down-regulation . Moreover , ∆(9)-THC caused a dose-dependent decrease in O60936 mRNA levels . The selective cannabinoid receptor P21554 antagonist AM251 ( 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1-piperidinyl-1H-pyrazole-3-carboxamide ) reduces both effects , suggesting that ∆(9)-THC activation of P21554 receptor is involved in the observed effects . These data show evidence of a cross-talk between O60936 and P21554 receptors , thus suggesting a possible interplay between cannabinoid and nociceptin/ O60936 system . [ Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture ] . In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10(-7)-10(-10) M it protected cells of sympathetic ganglia , neocortex and continues cell lines under damaging actions of H2O2 ( 0.0001 M ) , NH4CI ( 0.01 M ) and cooling . DB00086 essentially influenced the mode of damaging effect of DB00171 ( 0.001 M ) . Even a short-term exposition ( 20 min ) of PC12 cells with both proteins ( each in the concentration 10(-9) M ) led to sharp alterations in intracellular DB00171 - or Ca(2+)-activated proteolysis . In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation , improvement of cell adhesion , high survival rate , the increase in quantity and length of processes and their arborisation . Electronic microscopy established the character of structural rearrangements of nervous tissue cells ( neurons , astrocytes , oligodendrocytes ) , reflecting the protective action of plasminogen and streptokinase . In the presence of plasminogen and especially streptokinase , the total number of cultured glioma P13671 and neuroblastoma IMR-32 cells , the intracellular contents of protein , RNA and DNA increased several-fold . Addition of plasminogen promoted formation of processes by neuroblastoma cells , this suggests initiation of differentiation of cellular elements . In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells . These proteins did not cause transformation of PC12 enterochromaffine cells to neurons , though plasminogen facilitated it . P00747 addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium , and streptokinase did not lose its plasminogen-activating capacity . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . 123I-labeled AM251 : a radioiodinated ligand which binds in vivo to mouse brain cannabinoid P21554 receptors . We have investigated the binding of 123I-labeled N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methy l-1 H-pyrazole-3-carboxamide ( AM251 ) , an analog of the cannabinoid receptor antagonist SR141716A [ N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-me thyl-1 H-pyrazole-3-carboxamide ] in the mouse brain . Following intravenous injection , the peak whole-brain uptake of about 1 % of the administered activity occurred at about 2 h . By 8 h radioactivity in brain had declined to about half its peak value . High-performance liquid chromatographic analysis showed that > 70 % of radioactivity extracted from brain at 2 h was still present as [123I]AM251 . Co-injection of SR141716A inhibited the in vivo brain binding of [123I]AM251 dose dependently . At 2 mg/kg , the highest dose that could be tested , inhibition was 50 % at 2 h post-administration . The ED50 value calculated assuming that 2 mg/kg gave near-maximal inhibition was about 0.1 mg/kg . In contrast to the brain , radioactivity in other major organs ( blood , liver , kidney , heart and lung ) was little affected by SR141716A . The regional binding of [123I]AM251 in the brain was consistent with the published distribution of cannabinoid receptors in rat brain , in that the order was hippocampus , striatum > cerebellum > brain stem. delta 9- DB00470 co-administered intravenously at 10 mg/kg , a dose which induced catalepsy and decreased locomotor activity , decreased the 2 h brain uptake of [123I]AM251 by 10 % , but this was not significant ( P = 0.08 ) . In in vitro binding assays with mouse hippocampal membranes , tetrahydrocannabinol inhibited binding of [123I]AM251 with an IC50 value of about 700 nM , compared with about 0.2 nM for SR141716A . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . 2-arachidonoylglycerol , an endogenous cannabinoid receptor ligand , induces rapid actin polymerization in HL-60 cells differentiated into macrophage-like cells . Delta9- DB00470 , a major psychoactive constituent of marijuana , interacts with specific receptors , i.e. the cannabinoid receptors , thereby eliciting a variety of pharmacological responses . To date , two types of cannabinoid receptors have been identified : the P21554 receptor , which is abundantly expressed in the nervous system , and the CB2 receptor , which is predominantly expressed in the immune system . Previously , we investigated in detail the structure-activity relationship of various cannabinoid receptor ligands and found that 2-AG ( 2-arachidonoylglycerol ) is the most efficacious agonist . We have proposed that 2-AG is the true natural ligand for both the P21554 and CB2 receptors . Despite the potential physiological importance of 2-AG , not much information is available concerning its biological activities towards mammalian tissues and cells . In the present study , we examined the effect of 2-AG on morphology as well as the actin filament system in differentiated HL-60 cells , which express the CB2 receptor . We found that 2-AG induces rapid morphological changes such as the extension of pseudopods . We also found that it provokes a rapid actin polymerization in these cells . Actin polymerization induced by 2-AG was abolished when cells were treated with SR144528 , a CB2 receptor antagonist , and pertussis toxin , suggesting that the response was mediated by the CB2 receptor and G(i/o) . A phosphoinositide 3-kinase , Rho family small G-proteins and a tyrosine kinase were also suggested to be involved . Reorganization of the actin filament system is known to be indispensable for a variety of cellular events ; it is possible that 2-AG plays physiologically essential roles in various inflammatory cells and immune-competent cells by inducing a rapid actin rearrangement . Potential antipsychotic properties of central cannabinoid ( P21554 ) receptor antagonists . Delta(9)- DB00470 ( Delta(9)-THC ) , the principal psychoactive constituent of the Cannabis sativa plant , and other agonists at the central cannabinoid ( CB(1) ) receptor may induce characteristic psychomotor effects , psychotic reactions and cognitive impairment resembling schizophrenia . These effects of Delta(9)-THC can be reduced in animal and human models of psychopathology by two exogenous cannabinoids , cannabidiol ( DB09061 ) and SR141716 . DB09061 is the second most abundant constituent of Cannabis sativa that has weak partial antagonistic properties at the CB(1) receptor . DB09061 inhibits the reuptake and hydrolysis of anandamide , the most important endogenous CB(1) receptor agonist , and exhibits neuroprotective antioxidant activity . SR141716 is a potent and selective CB(1) receptor antagonist . Since both DB09061 and SR141716 can reverse many of the biochemical , physiological and behavioural effects of CB(1) receptor agonists , it has been proposed that both DB09061 and SR141716 have antipsychotic properties . Various experimental studies in animals , healthy human volunteers , and schizophrenic patients support this notion . Moreover , recent studies suggest that cannabinoids such as DB09061 and SR141716 have a pharmacological profile similar to that of atypical antipsychotic drugs . In this review , both preclinical and clinical studies investigating the potential antipsychotic effects of both DB09061 and SR141716 are presented together with the possible underlying mechanisms of action . [18F]MK-9470 PET measurement of cannabinoid P21554 receptor availability in chronic cannabis users . Δ(9) - DB00470 , the main psychoactive component of cannabis , exerts its central effects through activation of the cerebral type 1 cannabinoid ( P21554 ) receptor . Pre-clinical studies have provided evidence that chronic cannabis exposure is linked to decreased P21554 receptor expression and this is thought to be a component underlying drug tolerance and dependence . In this study , we make first use of the selective high-affinity positron emission tomography ( PET ) ligand [ ( 18 ) F ] MK-9470 to obtain in vivo measurements of cerebral P21554 receptor availability in 10 chronic cannabis users ( age = 26.0 ± 4.1 years ) . Each patient underwent [ ( 18 ) F ] MK-9470 PET within the first week following the last cannabis consumption . A population of 10 age-matched healthy subjects ( age = 23.0 ± 2.9 years ) was used as control group . Parametric modified standardized uptake value images , reflecting P21554 receptor availability , were calculated . Statistical parametric mapping and volume-of-interest ( VOI ) analyses of P21554 receptor availability were performed . Compared with controls , cannabis users showed a global decrease in P21554 receptor availability ( -11.7 percent ) . VOI-based analysis demonstrated that the P21554 receptor decrease was significant in the temporal lobe ( -12.7 percent ) , anterior ( -12.6 percent ) and posterior cingulate cortex ( -13.5 percent ) and nucleus accumbens ( -11.2 percent ) . Voxel-based analysis confirmed this decrease and regional pattern in P21554 receptor availability in cannabis users . These findings revealed that chronic cannabis use may alter specific regional P21554 receptor expression through neuroadaptive changes in P21554 receptor availability , opening the way for the examination of specific P21554 -cannabis addiction interactions which may predict future cannabis-related treatment outcome . Cannabinoid agonists in the treatment of blepharospasm -- a case report study . The benign essential blepharospasm is a subliminal form of primary torsion dystonia with still uncertain aetiology . It is characterized by involuntary convulsive muscle contractions of the M. orbicularis occuli , accompanied by unbearable pain of the cornea , eye bulb and the muscle itself . It has been suggested that blepharospasm is neurobiologically based on a dysfunction of the basal ganglia and an impairment of the dopamine neurotransmitter system . Therefore , therapy of blepharospasm contains administration of anticholinergic- and tranquillizing drugs as well as botulinum toxin as neuromuscular blocking agent . However serious side effects can be observed as well as failure of therapy . In the brain a dense co-localisation of cannabinoid ( P21554 ) and dopamine ( D2 ) -receptor was identified which had been associated with the influence of cannabinoids on the dopaminergic reward system . Additionally , it has been demonstrated that cannabinoids may have an impact on the central GABAergic and glutaminergic transmitter system and thus might be involved in the influence of movement control . In the present case we administered the cannabinoid receptor agonist Dronabinol ( Delta-9- DB00470 ) to a woman suffering from severe blepharospasm . Multiple treatments with botulinum toxin did not reveal a long-lasting beneficial effect . By contrast , treatment with 25 mg Dronabinol for several weeks improved the patients ' social life and attenuated pain perception remarkably . This case study demonstrates that the therapy with a cannabinoid agonist may provide a novel tool in the treatment of blepharospasm and maybe of other multifactorial related movement disorders . Increased mortality , hypoactivity , and hypoalgesia in cannabinoid P21554 receptor knockout mice . Delta9- DB00470 ( Delta9-THC ) , the major psychoactive ingredient in preparations of Cannabis sativa ( marijuana , hashish ) , elicits central nervous system ( CNS ) responses , including cognitive alterations and euphoria . These responses account for the abuse potential of cannabis , while other effects such as analgesia suggest potential medicinal applications . To study the role of the major known target of cannabinoids in the CNS , the P21554 cannabinoid receptor , we have produced a mouse strain with a disrupted P21554 gene . P21554 knockout mice appeared healthy and fertile , but they had a significantly increased mortality rate . They also displayed reduced locomotor activity , increased ring catalepsy , and hypoalgesia in hotplate and formalin tests . Delta9-THC-induced ring-catalepsy , hypomobility , and hypothermia were completely absent in P21554 mutant mice . In contrast , we still found Delta9-THC-induced analgesia in the tail-flick test and other behavioral ( licking of the abdomen ) and physiological ( diarrhea ) responses after Delta9-THC administration . Thus , most , but not all , CNS effects of Delta9-THC are mediated by the P21554 receptor . CNS effects of CB2 cannabinoid receptors : beyond neuro-immuno-cannabinoid activity . There are two well characterized cannabinoid receptors ( CBRs ) , P21554 -Rs and CB2-Rs , with other candidates , such as Q9Y2T6 , PPARs and vanilloid Q8NER1 ( Q8NER1 ) receptors , which are either activated by cannabinoids and/or endocannabinoids ( eCBs ) . The neuronal and functional expression of CB2-Rs in the brain has been much less well characterized in comparison with the expression of the ubiquitous P21554 -Rs . CB2-Rs were previously thought to be predominantly expressed in immune cells in the periphery and were traditionally referred to as peripheral CB2-Rs . We and others have now demonstrated the expression of CB2-Rs in neuronal , glial and endothelial cells in the brain , and this warrants a re-evaluation of the CNS effects of CB2-Rs . In the present review we summarize our current understanding of P34972 genomic structure , its polymorphic nature , subtype specificity , from mice to human subjects , and its variants that confer vulnerabilities to neuropsychiatric disorders beyond neuro-immuno-cannabinoid activity . Interactions between A-9THC and capsaicin on isolated lamb bladder detrusor . A number of studies have demonstrated that capsaicin , a capsicum alkaloid , can affect isolated bladder tissue with either a relaxation or a contraction , depending on the species , by acting on Q8NER1 receptors . In a previous work on isolated lamb detrusor , we demonstrated that capsaicin generally produces a relaxation of the tissue ; this relaxation seems to be mediated by P80511 . Endogenous cannabinoids , such as anandamide , produce some of their actions by stimulating Q8NER1 receptors and this seems to cause the release of peptides , e.g. P80511 . The aim of this work was to ascertain whether a cannabinoid , DB00470 ( delta-9THC ) , was able to interfere with the response of the isolated lamb detrusor to capsaicin . A-9THC , at concentrations between 1.6 x 10(-7) and 1.3 x 10(-6) M , displayed no activity on tissues . Instead , following delta-9THC , most of the tissues responded to capsaicin with a contraction that was abolished by atropine ( 9.0 x 10(-7) M ) . It has been reported that cannabinoids can inhibit the release of P80511 by stimulation of P21554 and CB2 cannabinoid receptors . Delta-9THC could act stimulating these receptors and thus inhibiting P80511 release and vesical relaxation . The muscle relaxing component removal could favour the contracting component , usually not active . Depolarization-induced suppression of excitation in murine autaptic hippocampal neurones . Depolarization-induced suppression of excitation and inhibition ( Q9UL01 and DSI ) appear to be important forms of short-term retrograde neuronal plasticity involving endocannabinoids ( eCB ) and the activation of presynaptic cannabinoid P21554 receptors . We report here that P21554 -dependent Q9UL01 can be elicited from autaptic cultures of excitatory mouse hippocampal neurones . Q9UL01 in autaptic cultures is both more robust and elicited with a more physiologically relevant stimulus than has been thus far reported for conventional hippocampal cultures . An additional requirement for autaptic Q9UL01 is filled internal calcium stores . Pharmacological experiments favour a role for 2-arachidonyl glycerol ( 2-AG ) rather than arachidonyl ethanolamide ( AEA ) or noladin ether as the relevant endocannabinoid to elicit Q9UL01 . In particular , the latter two compounds fail to reversibly inhibit EPSCs , a quality inconsistent with the role of bona fide eCB mediating Q9UL01 . Delta9- DB00470 ( delta9-THC ) fails to inhibit EPSCs , yet readily occludes both Q9UL01 and EPSC inhibition by a synthetic P21554 agonist , Q08050 55212-2 . With long-term exposure ( approximately 18 h ) , delta9-THC also desensitizes P21554 receptors . Lastly , a functional endocannabinoid transporter is necessary for the expression of Q9UL01 . Reinforcing and neurochemical effects of cannabinoid P21554 receptor agonists , but not cocaine , are altered by an adenosine A2A receptor antagonist . Several recent studies suggest functional and molecular interactions between striatal adenosine A(2A) and cannabinoid CB(1) receptors . Here , we demonstrate that A(2A) receptors selectively modulate reinforcing effects of cannabinoids . We studied effects of A(2A) receptor blockade on the reinforcing effects of DB00470 ( THC ) and the endogenous CB(1) receptor ligand anandamide under a fixed-ratio schedule of intravenous drug injection in squirrel monkeys . A low dose of the selective adenosine A(2A) receptor antagonist MSX-3 ( 1 mg/kg ) caused downward shifts of THC and anandamide dose-response curves . In contrast , a higher dose of MSX-3 ( 3 mg/kg ) shifted THC and anandamide dose-response curves to the left . MSX-3 did not modify cocaine or food pellet self-administration . Also , MSX-3 neither promoted reinstatement of extinguished drug-seeking behavior nor altered reinstatement of drug-seeking behavior by non-contingent priming injections of THC . Finally , using in vivo microdialysis in freely-moving rats , a behaviorally active dose of MSX-3 significantly counteracted THC-induced , but not cocaine-induced , increases in extracellular dopamine levels in the nucleus accumbens shell . The significant and selective results obtained with the lower dose of MSX-3 suggest that adenosine A(2A) antagonists acting preferentially at presynaptic A(2A) receptors might selectively reduce reinforcing effects of cannabinoids that lead to their abuse . However , the appearance of potentiating rather than suppressing effects on cannabinoid reinforcement at the higher dose of MSX-3 would likely preclude the use of such a compound as a medication for cannabis abuse . DB00640 A(2A) antagonists with more selectivity for presynaptic versus postsynaptic receptors could be potential medications for treatment of cannabis abuse . Possible involvement of endocannabinoids in the increase of morphine consumption in maternally deprived rat . Whether adolescent exposure to chronic DB00470 ( THC ) facilitates progression to opioid consumption is still controversial . In a maternal deprivation model ( 3 h daily from postnatal day 1-14 ) , we previously reported that adolescent exposure to chronic THC blocks morphine dependence in maternally deprived ( D ) rats . Owing to the existence of a functional cross-interaction between the opioid and cannabinoid systems in reward , we evaluated if the vulnerability to opiate reward in D rats , may involve an alteration of the endocannabinoid system . Anandamide and 2-arachidonoylglycerol ( 2-AG ) , were quantified in the striatum and mesencephalon of adolescent and adult D and non-deprived ( animal facility rearing , AFR ) rats by isotope dilution liquid chromatography-mass spectrometry . Oral morphine self-administration behavior was analyzed for 14 weeks , 24 days after chronic injection of the cannabinoid P21554 receptor antagonist/inverse agonist , SR141716A ( 3 mg/kg ) for 2 weeks during adolescence ( P01160 35-48 ) . Adolescent D rats exhibited higher basal levels of anandamide than adolescent AFR rats in the nucleus accumbens ( 38 % ) , the caudate-putamen nucleus ( 62 % ) and the mesencephalon ( 320 % ) , whereas adult D rats showed an increase of anandamide and 2-AG levels in the nucleus accumbens ( 50 % and 24 % , respectively ) and of 2-AG in the caudate-putamen nucleus ( 48 % ) , compared to adult AFR rats . Chronic administration of SR141716A to adolescent D rats blocked the escalation behavior in the morphine consumption test . Our data suggest that altered brain endocannabinoid levels may contribute to the escalation behavior in the morphine consumption test in a maternal deprivation model . Neonatal melanocortin receptor agonist treatment reduces play fighting and promotes adult attachment in prairie voles in a sex-dependent manner . The melanocortin receptor ( P08235 ) system has been studied extensively for its role in feeding and sexual behavior , but effects on social behavior have received little attention . α-MSH interacts with neural systems involved in sociality , including oxytocin , dopamine , and opioid systems . Acute melanotan-II ( MTII ) , an MC3/4R agonist , potentiates brain oxytocin ( OT ) release and facilitates OT-dependent partner preference formation in socially monogamous prairie voles . Here we examined the long-term impact of early-life P08235 stimulation on hypothalamic neuronal activity and social development in prairie voles . Male and female voles were given daily subcutaneous injections of 10 mg/kg MTII or saline between postnatal days ( P01160 ) 1-7 . Neonatally-treated males displayed a reduction in initiated play fighting bouts as juveniles compared to control males . Neonatal exposure to MTII facilitated partner preference formation in adult females , but not males , after a brief cohabitation with an opposite-sex partner . Acute MTII injection elicited a significant burst of the immediate early gene P18146 immunoreactivity in hypothalamic OT , vasopressin , and corticotrophin releasing factor neurons , when tested in P01160 6-7 animals . Daily neonatal treatment with 1 mg/kg of a more selective , brain penetrant P32245 agonist , PF44687 , promoted adult partner preferences in both females and males compared with vehicle controls . Thus , developmental exposure to P08235 agonists lead to a persistent change in social behavior , suggestive of structural or functional changes in the neural circuits involved in the formation of social relationships . delta 9- DB00470 increases activity of tyrosine hydroxylase in cultured fetal mesencephalic neurons . The exposure of pregnant rats to delta 9-tetrahydrocannabinol ( delta 9-THC ) , the main psychoactive constituent of Cannabis sativa , during gestation and lactation , affects the gene expression and the activity of tyrosine hydroxylase ( TH ) in the brain of their offspring , measured at fetal and early postnatal ages , when the expression of this enzyme plays an important role in neural development . In the present article , we have examined whether delta 9-THC is able to affect TH activity in cultured mesencephalic neurons obtained from fetuses at gestational d 14 . Thus , TH activity increased approximately twofold in cells obtained from naive fetuses when exposed for 24 h to medium containing delta 9-THC . In addition , TH activity was also approx twofold higher in cells obtained from fetuses exposed daily to delta 9-THC from d 5 of gestation than in cells obtained from control fetuses , when both were exposed to basal media . This effect of delta 9-THC on TH activity seems to be produced via the activation to cannabinoid receptors , in particular the P21554 subtype , which would presumably be located in these cells . This is because the exposure to medium containing both delta 9-THC and SR141716A , a specific antagonist for P21554 receptors , abolished the effect observed with delta 9-THC alone . SR141716A alone was without effect on TH activity . Collectively , our results support the notion that delta 9-THC increased TH activity in cultured mesencephalic neurons , as previously observed in vivo , and that this effect was produced by activation of P21554 receptors , which seem to be operative at these early ages . All this points to a role for the endogenous cannabimimetic system in brain development . P21554 receptor knockout mice show similar behavioral modifications to wild-type mice when enkephalin catabolism is inhibited . Behavioral and biochemical studies have suggested a functional link between the endogenous cannabinoid and opioid systems . Different hypotheses have been proposed to explain the interactions between opioid and cannabinoid systems such as a common pathway stimulating the dopaminergic system , a facilitation of signal-transduction- and/or a cannabinoid-induced enhancement of opioid peptide release . However , at this time , all the studies have been performed with exogenous agonists ( DB00470 or morphine ) , leading to a generally excessive stimulation of receptors normally stimulated by endogenous effectors ( anandamide or opioid peptides ) in various brain structures . To overcome this problem , we have measured various behavioral responses induced by the stimulation of the endogenous opioid system using the dual inhibitor of enkephalin-degrading enzymes , RB101 , in P21554 receptor knockout mice . Thus , analgesia , locomotor activity , anxiety and antidepressant-like effects were measured after RB101 administration ( 80 and 120 mg/kg i.p. or 10 mg/kg , i.v. ) in P21554 receptor knockout mice and their wild-type littermates . In all the experiments , inhibition of enkephalin catabolism produced similar modifications in behavior observed in P21554 knockout and wild-type mice . These results suggest limited physiological interaction between cannabinoid and opioid systems . DB00470 -induced apoptosis of cultured cortical neurones is associated with cytochrome c release and caspase-3 activation . Delta(9)-tetrahydrocannabinol ( THC ) , the principal psychoactive component of marijuana , is associated with impaired cognition and altered cortical function . THC transduces its central effects via activation of the G-protein linked cannabinoid receptor P21554 . In this study we report that THC induces morphological degenerative changes in cultured cortical neurones , such as membrane blebbing and formation of apoptotic bodies , that are consistent with the apoptotic pathway of cell death . The THC-induced apoptosis was blocked by the P21554 receptor antagonist AM251 and pertussis toxin ( PTX ) , suggesting that this effect of THC involves receptor-mediated activation of the G-protein subtypes G(i) or G(o) . THC also promoted translocation of mitochondrial cytochrome c to the cytosol and increased the activity of the cysteine protease caspase-3 , in a PTX-sensitive manner . The results from this study suggest that coupling of THC to a PTX-sensitive G-protein promotes cytochrome c release , caspase-3 activation and subsequent degeneration of cultured cortical neurones . This apoptotic pathway may underlie the compromised neuronal function that is associated with marijuana usage . P29323 signalling pathway is not involved in PSA- P13591 -dependent alterations of hippocampal plasticity evoked by P21554 receptor activation . The present study investigated the potential role of the extracellular signal-regulated kinase ( P29323 ) pathway in the alternation of polysialylated neural cell adhesion molecule ( PSA- P13591 ) expression and proliferation rates in the dentate gyrus ( DG ) evoked by activation of the P21554 receptor . When given at a dose of 0.1 mg/kg , the P21554 receptor agonist , 3-(1,1-dimethylheptyl)-11-hydroxy-Delta(8)-tetrahydrocannabinol ( HU-210 ) , increased the levels of the phosphorylated forms of P29323 ( pERK1 and pERK2 ) in the hippocampus when measured 30 min after injection . This HU-210-induced effect was inhibited by alpha-{amino[(4-aminophenyl)thio]methylene}-2-(trifluoromethyl) benzeneacetonitrile ( SL327 , 30 mg/kg ) - an inhibitor of mitogen-activated protein kinase kinase ( Q02750 /2 ) , the upstream kinase of P29323 - given 1 h before HU-210 administration . Additionally , SL327 alone significantly attenuated the basal level of both pERK1 and pERK2 . HU-210 ( 0.1 mg/kg ) decreased the number of PSA- P13591 -immunoreactive ( IR ) cells but did not affect the rate of proliferation , which was analyzed as the number of Ki-67-IR cells measured in the DG 2 days after HU-210 administration . The data indicated that SL327 ( 30 mg/kg ) alone decreased the number of PSA- P13591 -IR cells 2 days after treatment . Joint administration of SL327 and HU-210 decreased the number of PSA- P13591 cells more robustly than did the administration of either alone . In addition , SL327 did not decrease the number of Ki-67-IR cells , while pretreatment with SL327 1 h before HU-210 administration did . These results suggest that stimulation of the P29323 cascade caused by P21554 receptor activation is not involved in hippocampal plasticity governed by PSA- P13591 expression . Antiemetic and motor-depressive actions of CP55,940 : cannabinoid P21554 receptor characterization , distribution , and G-protein activation . Dibenzopyran ( Delta(9)-tetrahydrocannabinol ) and aminoalkylindole [ R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrolol[1,2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl) methanone mesylate ; ( WIN55,212-2 ) ] cannabinoids suppress vomiting produced by cisplatin via cannabinoid CB(1) receptors . This study investigates the antiemetic potential of the " nonclassical " cannabinoid CP55,940 [ 1alpha,2beta-(R)-5alpha ] -(-)-5-(1,1-dimethyl)-2- [ 5-hydroxy-2-(3-hydroxypropyl) cyclohexyl-phenol ] against cisplatin-induced vomiting and assesses the presence and functionality of cannabinoid CB(1) receptors in the least shrew ( Cryptotis parva ) brain . CP55,940 ( 0.025-0.3 mg/kg ) reduced both the frequency of cisplatin-induced emesis ( ID(50)=0.025 mg/kg ) and the percentage of shrews vomiting ( ID(50)=0.09 mg/kg ) . CP55,940 also suppressed shrew motor behaviors ( ID(50)=0.06- 0.21 mg/kg ) at such doses . The antiemetic and motor-suppressant actions of CP55,940 were countered by SR141716A [ N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)- DB01213 -3-carboxamide ] , indicating both effects are cannabinoid CB(1) receptor-mediated . Autoradiographic studies with [ 3H ] -SR141716A and [ 35S ] -GTPgammaS binding revealed that the distribution of the cannabinoid CB(1) receptor and its activation pattern are similar to rodent brain and significant levels are present in brain loci ( e.g. , nucleus tractus solitarius ( P30990 ) ) that control emesis . The affinity rank order of structurally diverse cannabinoid ligands for cannabinoid CB(1) receptor in shrew brain is similar to rodent brain : HU-210=CP55,940=SR141716A >/= WIN55,212-2 >/= DB00470 > methanandamide=HU-211=cannabidiol=2-arachidonoylglycerol . This affinity order is also similar and is highly correlated to the cannabinoid EC(50) potency rank order for GTPgammaS stimulation except WIN55,212-2 and DB00470 potency order were reversed . The affinity and the potency rank order of tested cannabinoids were significantly correlated with their antiemetic ID(50) potency order against cisplatin-induced vomiting ( CP55,940 > WIN55,212-2= DB00470 ) as well as emesis produced by 2-arachidonoylglycerol or SR141716A ( CP55,940 > WIN55,212-2 > DB00470 ) . Evidence for an interaction between P21554 cannabinoid and melanocortin P08235 -4 receptors in regulating food intake . P32245 ( MCR4 ) and CB(1) cannabinoid receptors independently modulate food intake . Although an interaction between the cannabinoid and melanocortin systems has been found in recovery from hemorrhagic shock , the interaction between these systems in modulating food intake has not yet been examined . The present study had two primary purposes : 1 ) to examine whether the cannabinoid and melanocortin systems act independently or synergistically in suppressing food intake ; and 2 ) to determine the relative position of the CB(1) receptors in the chain of control of food intake in relation to the melanocortin system . Rats were habituated to the test environment and injection procedure and then received intracerebroventicular injections of various combinations of the MCR4 receptor antagonist JKC-363 , the CB(1) receptor agonist Delta(9)-tetrahydrocannabinol , the MCR4 receptor agonist alpha-MSH , or the cannabinoid CB(1) receptor antagonist SR 141716 . Food intake and locomotor activity were then recorded for 120 min . When administrated alone , SR 141716 and alpha-MSH dose-dependently attenuated baseline feeding , whereas sub-anorectic doses of SR 141716 and alpha-MSH synergistically attenuated baseline feeding when combined . Delta(9)- DB00470 -induced feeding was not blocked by alpha-MSH , whereas SR 141716 dose-dependently attenuated JKC-363-induced feeding . Locomotor activity was not significantly affected by any drug treatment , suggesting that the observed effects on feeding were not due to a nonspecific reduction in motivated behavior . These findings revealed a synergistic interaction between the cannabinoid and melanocortin systems in feeding behavior . These results further suggested that CB(1) receptors are located downstream from melanocortin receptors and CB(1) receptor signaling is necessary to prevent the melanocortin system from altering food intake . Novel MEK inhibitor trametinib and other retinoblastoma gene ( RB ) -reactivating agents enhance efficacy of 5-fluorouracil on human colon cancer cells . Chemotherapy for colorectal cancer has become more complicated and diversified with the appearance of molecular-targeting agents . DB00544 ( DB00544 ) has been a mainstay of chemotherapy for colorectal cancer , but it is still unknown whether the combining of DB00544 with novel molecular-targeting agents is effective . P04818 ( TS ) is a direct target of DB00544 , and the low TS level has been generally supposed to sensitize DB00544 's efficacy . We therefore hypothesized that RB-reactivating agents could enhance the efficacy of DB00544 , because the RB-reactivating agents could suppress the function of transcription factor E2F of TS gene promoter . We used three RB-reactivating agents , trametinib/GSK1120212 ( MEK inhibitor ) , fenofibrate ( Q07869 α agonist ) , and LY294002 ( PI3K inhibitor ) , with DB00544 against human colon cancer HT-29 and HCT15 cells . DB08911 induced p15 and p27 expression and reduced cyclin D1 levels in HT-29 cells . Fenofibrate also dephosphorlated P27361 /2 and reduced cyclin D1 levels in HT-29 cells . LY294002 induced p27 expression in HCT15 cells . All three agents caused dephosphorylation of RB protein and P55008 -phase arrest with a reduction of TS expression . As a consequence , the combination of DB00544 with each of the agents resulted in a significant decrease of colony numbers in HT-29 or HCT15 cells . These results suggest " RB-reactivation therapy " using molecular-targeting agents to be a new strategy for DB00544 -based chemotherapy . In particular , we strongly expect trametinib , which was discovered in Japan and was recently submitted to FDA for approval , to be used together with established regimens for colorectal cancer . Replication of obesity and diabetes-related SNP associations in individuals from Yucatán , México . The prevalence of type 2 diabetes ( T2D ) is rising rapidly and in Mexicans is ~19 % . T2D is affected by both environmental and genetic factors . Although specific genes have been implicated in T2D risk few of these findings are confirmed in studies of Mexican subjects . Our aim was to replicate associations of 39 single nucleotide polymorphisms ( SNPs ) from 10 genes with T2D-related phenotypes in a community-based Mexican cohort . Unrelated individuals ( n = 259 ) living in southeastern Mexico were enrolled in the study based at the University of Yucatan School of Medicine in Merida . Phenotypes measured included anthropometric measurements , circulating levels of adipose tissue endocrine factors ( leptin , adiponectin , pro-inflammatory cytokines ) , and insulin , glucose , and blood pressure . Association analyses were conducted by measured genotype analysis implemented in SOLAR , adapted for unrelated individuals . SNP Minor allele frequencies ranged from 2.2 to 48.6 % . Nominal associations were found for P21554 , Q8IWU4 , GCK , and P29120 SNPs with systolic blood pressure , insulin and glucose , and for P21554 , Q8IWU4 , Q14654 , and P29120 SNPs with adiponectin and leptin ( p < 0.05 ) . P-values greater than 0.0014 were considered significant . Association of SNPs rs10485170 of P21554 and rs5215 of Q14654 with adiponectin and leptin , respectively , reached near significance ( p = 0.002 ) . Significant association ( p = 0.001 ) was observed between plasma leptin and rs5219 of Q14654 . DB01197 attenuates matrix metalloproteinase-2 and -9 in monocrotaline-induced right ventricular hypertrophy in rats . Little is known about the influence of angiotensin converting enzyme ( P12821 ) inhibitors on matrix metalloproteinase ( MMP ) in right ventricular remodeling . We investigated the effect of captopril , an P12821 inhibitor , on P08253 and P14780 in monocrotaline-induced right ventricular hypertrophy . Six-week-old male Wistar rats were injected intraperitoneally with monocrotaline ( 60 mg/kg ) or saline . The rats were administrated captopril ( 30 mg/kg per day ) or a vehicle orally for 24 days from the day of monocrotaline injection . At day 25 , echocardiography was performed and hearts were excised . Expressions and activities of P08253 and P14780 were measured by Western blotting and by gelatin zymography , respectively . In monocrotaline-injected rats , right ventricular weight/tail length ratio increased significantly . Histological analysis revealed cardiomyocyte hypertrophy and fibrosis in right ventricular sections . Echocardiography showed right ventricular dysfunction compared with saline-injected rats . The right ventricular hypertrophy , fibrosis , and dysfunction were inhibited by captopril . However , captopril did not attenuate an increase in pulmonary artery pressure . P08253 and P14780 expressions and activities in right ventricles increased significantly in monocrotaline-injected rats and captopril inhibited them . These findings indicate that captopril attenuates the development of monocrotaline-induced right ventricular hypertrophy in association with inhibition of P08253 and P14780 in rats . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . Cannabinoids against pain . Efficacy and strategies to reduce psychoactivity : a clinical perspective . The clinical use of cannabinoids is currently a topic of interest not exclusively , but most importantly , concerning different areas of pain therapy . One of the major obstacles in developing clinically acceptable compounds is the cannabimimetic side-effect profile of DB00470 ( THC ) and other cannabinoids . This article gives a brief overview of the endocannabinoid system , its components and functions and explains the current approaches to avoiding cannabimimetic side effects by separating them from the therapeutic effects . One of these approaches is the addition of cannabidiol ( DB09061 ) as well as the use of preparations suitable for oromucosal application . Also cannabinoids , which primarily stimulate peripheral cannabinoid-1 ( P21554 ) receptors or selectively cannabinoid-2 ( CB2 ) receptors , can further separate analgesic activity from cannabimimetic activity . Local or topical modes of application are another attempt aiming in the same direction . Modulating the endogenous cannabinoid tone ( via the inhibition of endocannabinoid-metabolising enzymes ) is another strategy . The combination of THC in low , non-psychoactive doses with opioids has a synergistic effect and reduces opioid tolerance effects . Available data from these approaches are summarised and their more and less promising aspects are discussed . Cannabis-induced cytotoxicity in leukemic cell lines : the role of the cannabinoid receptors and the MAPK pathway . Delta9- DB00470 ( THC ) is the active metabolite of cannabis . THC causes cell death in vitro through the activation of complex signal transduction pathways . However , the role that the cannabinoid 1 and 2 receptors ( P21554 -R and CB2-R ) play in this process is less clear . We therefore investigated the role of the CB-Rs in mediating apoptosis in 3 leukemic cell lines and performed microarray and immunoblot analyses to establish further the mechanism of cell death . We developed a novel flow cytometric technique of measuring the expression of functional receptors and used combinations of selective P21554 -R and CB2-R antagonists and agonists to determine their individual roles in this process . We have shown that THC is a potent inducer of apoptosis , even at 1 x IC(50) ( inhibitory concentration 50 % ) concentrations and as early as 6 hours after exposure to the drug . These effects were seen in leukemic cell lines ( CEM , HEL-92 , and HL60 ) as well as in peripheral blood mononuclear cells . Additionally , THC did not appear to act synergistically with cytotoxic agents such as cisplatin . One of the most intriguing findings was that THC-induced cell death was preceded by significant changes in the expression of genes involved in the mitogen-activated protein kinase ( MAPK ) signal transduction pathways . Both apoptosis and gene expression changes were altered independent of p53 and the CB-Rs . CB2 cannabinoid receptor agonist JWH-015 modulates human monocyte migration through defined intracellular signaling pathways . Recruitment of leukocytes to inflammatory sites is crucial in the pathogenesis of chronic inflammatory diseases . The aim of this study was to investigate if activation of CB2 cannabinoid receptors would modulate the chemotactic response of human monocytes . Human monocytes treated with the CB2 agonist JWH-015 for 12-18 h showed significantly reduced migration to chemokines P13500 and P10147 , associated with reduced mRNA and surface expression of their receptors P41597 and P32246 . The induction of P05362 in response to P01579 was inhibited by JWH-015 . Moreover , JWH-015 cross-desensitized human monocytes for migration in response to P13500 and P10147 by its own chemoattractant properties . The CB2-selective antagonist SR-144528 , but not the P21554 antagonist SR-147778 , reversed JWH-015-induced actions , whereas the CB2 agonist JWH-133 mimicked the effects of JWH-015 . The investigation of underlying pathways revealed the involvement of phosphatidylinositol 3-kinase/Akt and P27361 /2 but not p38 MAPK . In conclusion , selective activation of CB2 receptors modulates chemotaxis of human monocytes , which might have crucial effects in chronic inflammatory disorders such as atherosclerosis or rheumatoid arthritis . Pancreatic gene expression during the initiation of acute pancreatitis : identification of P18146 as a key regulator . We hypothesized that genes expressed in pancreatic acinar cells during the initiation of acute pancreatitis determine the severity of the disease . Therefore , we utilized microarrays to identify those genes commonly induced in rat pancreatic acinar cells within 1-4 h in two in vivo models , caerulein and taurocholate administration . This strategy yielded 51 known genes representing a complex array of molecules , including those that are likely to either reduce or increase the severity of the disease . Novel genes identified in the current study included P18847 , BRF1 , C/EBPbeta , P80511 , P18146 , ephrinA1 , villin2 , ferredoxin , latexin , lipocalin , P28562 , NGFI-B , RhoA , tissue factor ( TF ) , and syndecan . To validate these microarray results , the role of P18146 was further investigated using quantitative RT-PCR , Western blotting , and immunocytochemistry . P18146 expression occurred within acinar cells and correlated with the development of caerulein-induced acute pancreatitis in rats . Furthermore , the levels of the inflammation-related genes P13500 , P05121 , TF , P05231 , and P05362 and the extent of lung inflammation were reduced during the initiation of caerulein-induced acute pancreatitis in P18146 -deficient mice . Thus this study identified P18146 and several other novel genes likely to be important in the development and severity of acute pancreatitis . Anti-inflammatory activity of topical THC in DNFB-mediated mouse allergic contact dermatitis independent of P21554 and CB2 receptors . BACKGROUND : ∆(9) - DB00470 ( THC ) , the active constituent of Cannabis sativa , exerts its biological effects in part through the G-protein-coupled P21554 and CB2 receptors , which were initially discovered in brain and spleen tissue , respectively . However , THC also has P21554 /2 receptor-independent effects . Because of its immune-inhibitory potential , THC and related cannabinoids are being considered for the treatment of inflammatory skin diseases . Here we investigated the mechanism of the anti-inflammatory activity of THC and the role of P21554 and CB2 receptors . METHODS : We evaluated the impact of topically applied THC on DNFB-mediated allergic contact dermatitis in wild-type and P21554 /2 receptor-deficient mice . We performed immunohistochemical analyses for infiltrating immune cells and studied the influence of THC on the interaction between T cells , keratinocytes and myeloid immune cells in vitro . RESULTS : Topical THC application effectively decreased contact allergic ear swelling and myeloid immune cell infiltration not only in wild-type but also in P21554 /2 receptor-deficient mice . We found that THC ( 1 ) inhibited the production of IFNγ by T cells , ( 2 ) decreased the production of P13500 and of IFNγ-induced P80075 and CXL10 by epidermal keratinocytes and ( 3 ) thereby limited the recruitment of myeloid immune cells in vitro in a P21554 /2 receptor-independent manner . CONCLUSIONS : Topically applied THC can effectively attenuate contact allergic inflammation by decreasing keratinocyte-derived pro-inflammatory mediators that orchestrate myeloid immune cell infiltration independent of P21554 /2 receptors . This has important implications for the future development of strategies to harness cannabinoids for the treatment of inflammatory skin diseases . Influence of convolution filtering on coronary plaque attenuation values : observations in an ex vivo model of multislice computed tomography coronary angiography . Attenuation variability ( measured in Hounsfield Units , HU ) of human coronary plaques using multislice computed tomography ( MSCT ) was evaluated in an ex vivo model with increasing convolution kernels . MSCT was performed in seven ex vivo left coronary arteries sunk into oil followingthe instillation of saline ( 1/infinity ) and a 1/50 solution of contrast material ( 400 mgI/ml iomeprol ) . Scan parameters were : slices/collimation , 16/0.75 mm ; rotation time , 375 ms . Four convolution kernels were used : b30f-smooth , b36f-medium smooth , b46f-medium and b60f-sharp . An experienced radiologist scored for the presence of plaques and measured the attenuation in lumen , calcified and noncalcified plaques and the surrounding oil . The results were compared by the Q9UNW9 test and correlated with Pearson 's test . The signal-to-noise ratio ( SNR ) and contrast-to-noise ratio ( P21554 ) were calculated . The mean attenuation values were significantly different between the four filters ( p < 0.0001 ) in each structure with both solutions . After clustering for the filter , all of the noncalcified plaque values ( 20.8 +/- Q04695 , 14.2 +/- 35.8 , 14.0 +/- 32.0 , 3.2 +/- 32.4 HU with saline ; 74.7 +/- 66.6 , 68.2 +/- 63.3 , 66.3 +/- 66.5 , 48.5 +/- 60.0 HU in contrast solution ) were significantly different , with the exception of the pair b36f-b46f , for which a moderate-high correlation was generally found . Improved SNRs and CNRs were achieved by b30f and b46f . The use of different convolution filters significantly modifief the attenuation values , while sharper filtering increased the calcified plaque attenuation and reduced the noncalcified plaque attenuation . Construction of a steric map of the binding pocket for cannabinoids at the cannabinoid receptor . In order to gain information about the topology of the brain cannabinoid receptor ( P21554 ) , a Receptor Steric ( RS ) Map for cannabinoids at this receptor was calculated . The classical cannabinoids (-)-11-hydroxy- DB00470 ( P04264 = 210 +/- 56 nM ) , (-)-9-nor-9-beta-hydroxy-hexahydrocannabinol ( P04264 = 124 +/- 17 nM ) , nabilone ( P04264 = 120 +/- 13 nM ) , and the non-classical cannabinoid , CP-55,244 ( P04264 = 1.4 +/- .3 nM ) were used as template molecules . The RS map was obtained as the union of the van der Waals ' volumes of only those accessible conformers identified by P08253 calculations that were able to clear a region of steric interference at the P21554 receptor previously characterized by us [ Reggio , P.H. , Panu , A.M. and Miles , S. ( 1993 ) , J. Med. Chem. , 36 , 1761-1771 ] . The utility of the RS Map was explored by screening the accessible conformers of the classical cannabinoid , cannabinol ( CBN ) , ( P04264 = 3200 +/- 450 nM ) , for its ability to fit within the RS map . Only the global minimum energy conformer of CBN ( 53.2 % abundance at 298K ) was able to fit within the RS map . These results imply that one reason for the reduced affinity of CBN may be that only 53.2 % of CBN molecules are shaped properly to fit in the binding pocket for cannabinoids at the P21554 receptor . Tolerance to chronic DB00470 ( Δ⁹-THC ) in rhesus macaques infected with simian immunodeficiency virus . Although Δ⁹-THC has been approved to treat anorexia and weight loss associated with AIDS , it may also reduce well-being by disrupting complex behavioral processes or enhancing HIV replication . To investigate these possibilities , four groups of male rhesus macaques were trained to respond under an operant acquisition and performance procedure , and administered vehicle or Δ⁹-THC before and after inoculation with simian immunodeficiency virus ( SIV(mac251) , 100 TCID₅₀/ml , i.v. ) . Prior to chronic Δ⁹-THC and SIV inoculation , 0.032-0.32 mg/kg of Δ⁹-THC produced dose-dependent rate-decreasing effects and small , sporadic error-increasing effects in the acquisition and performance components in each subject . Following 28 days of chronic Δ⁹-THC ( 0.32 mg/kg , i.m. ) or vehicle twice daily , DB00470 -treated subjects developed tolerance to the rate-decreasing effects , and this tolerance was maintained during the initial 7-12 months irrespective of SIV infection ( i.e. , +THC/-SIV , +THC/+SIV ) . Q8N1N2 necropsy was performed on all SIV subjects an average of 329 days post-SIV inoculation , with postmortem histopathology suggestive of a reduced frequency of CNS pathology as well as opportunistic infections in DB00470 -treated subjects . Chronic Δ⁹-THC also significantly reduced CB-1 and P34972 receptor levels in the hippocampus , attenuated the expression of a proinflammatory cytokine ( P13500 ) , and did not increase viral load in plasma , cerebrospinal fluid , or brain tissue compared to vehicle-treated subjects with SIV . Together , these data indicate that chronic Δ⁹-THC produces tolerance to its behaviorally disruptive effects on complex tasks while not adversely affecting viral load or other markers of disease progression during the early stages of infection . Arachidonyl ethanolamide induces apoptosis of uterine cervix cancer cells via aberrantly expressed vanilloid receptor-1 . OBJECTIVES : Delta(9)- DB00470 , the active agent of Cannabis sativa , exhibits well-documented antitumor properties , but little is known about the possible effects mediated by endogenous cannabinoids on human tumors . In the present study , we analyzed the effect of arachidonyl ethanolamide ( AEA ) on cervical carcinoma ( CxCa ) cell lines . METHODS : To assess the sensitivity of CxCa cells to AEA , we selected three cell lines that were exposed to increasing doses of AEA with or without antagonists to receptors to AEA . DNA fragmentation and caspase-7 activity were used as apoptosis markers . The expression of receptors to AEA were analyzed in CxCa cell lines as well as CxCa biopsies . RESULTS : The major finding was that AEA induced apoptosis of CxCa cell lines via aberrantly expressed vanilloid receptor-1 , whereas AEA binding to the classical P21554 and CB2 cannabinoid receptors mediated a protective effect . Furthermore , unexpectedly , a strong expression of the three forms of AEA receptors was observed in ex vivo CxCa biopsies . CONCLUSIONS : Overall , these data suggest that the specific targeting of Q8NER1 by endogenous cannabinoids or synthetic molecules offers attractive opportunities for the development of novel potent anticancer drugs . Effects of DB00470 on human immune function and host defense . This review examines evidence that delta(9)-tetrahydrocannabinol ( THC ) can regulate and suppress human immune responses . Leukocytes express both cannabinoid receptor type 1 ( P21554 ) and cannabinoid receptor type 2 ( CB2 ) , and levels of mRNA encoding for them are increased in peripheral blood leukocytes obtained from marijuana smokers , suggesting cannabinoid receptor activation in vivo . Exposure of human T-cells to THC suppresses their proliferation , inhibits the release of interferon-gamma , and skews the balance of T-helper cytokines towards a type 2 response . The majority of these effects are CB2 receptor-dependent . Consistent with an impact of THC on cell-mediated immunity , alveolar macrophages ( AMs ) recovered from the lungs of marijuana smokers are suppressed in their ability to release pro-inflammatory cytokines and nitric oxide ( NO ) , and kill bacteria . Macrophage function is restored by treatment with interferon-gamma , a type 1 cytokine . Habitual exposure to THC appears capable of impacting on human cell-mediated immunity and host defense . The cannabinoid P21554 antagonists SR 141716A and AM 251 suppress food intake and food-reinforced behavior in a variety of tasks in rats . Cannabinoid P21554 receptor agonists , including DB00470 ( Delta 9-THC ) ( the main psychoactive ingredient in marijuana ) have been shown to increase feeding in rats and humans . Conversely , it has been reported that acute administration of the P21554 receptor antagonist SR 141716A reduces food intake in rats . Based upon this observation , it has been suggested that P21554 antagonists could be useful as appetite suppressant drugs . The present studies were designed to provide a detailed examination of the effects of P21554 antagonists on food intake across a range of paradigms . Two P21554 antagonists ( SR 141716A and AM 251 ) were administered to rats trained on fixed-ratio schedules with two different ratio requirements ( fixed-ratio 1 and fixed-ratio 5 ) . Both drugs produced a dose-dependent decrease in lever pressing , and had a relatively long duration of action ( T1/2 : SR 141716A , 15.1 h ; AM 251 , 22.0 h ) . Furthermore , intake of three diets with differing macronutrient composition ( lab chow , high fat , high carbohydrate ) was studied . Both drugs significantly suppressed intake of all three foods , and there were no significant interactions between drug dose and diet type . These findings support the hypothesis that P21554 receptor antagonists could be useful pharmacological tools for the suppression of appetite . Opposing control of cannabinoid receptor stimulation on amyloid-beta-induced reactive gliosis : in vitro and in vivo evidence . Beside cytotoxic mechanisms impacting on neurons , amyloid beta ( A beta ) -induced astroglial activation is operative in Alzheimer 's disease brain , suggesting that persistent inflammatory response may have a role in the illness and that positive results may be achieved by curbing the astroglial reaction . Because the role of the endocannabinoid system could represent a promising field of research , the present study conducted in vitro and in vivo experiments to assess this system . P13671 rat astroglioma cells were challenged with 1 microg/ml A beta 1-42 in the presence or absence of selective agonists and antagonists of cannabinoid (CB)1 and CB2 receptors . Furthermore , rats were inoculated into the frontal cortex with 30 ng of A beta 1-42 and were i.p. administered with 5 mg/kg of the same substances . Immunohistochemical and biochemical findings revealed that selective agonism at P21554 and antagonism at CB2 receptors was able to blunt A beta-induced reactive astrogliosis with subsequent overexpression of glial fibrillary acidic protein and P04271 protein . Moreover , A beta provoked down-regulation of P21554 receptors together with a reduction of anandamide concentration , whereas CB2 receptors were up-regulated and 2-arachidonoyl glycerol concentration was increased . Finally , to our knowledge , the current study is the first showing that interactions at cannabinoid receptors result in a dual regulation of A beta-induced reactive astrogliosis . The data support the assumption that compounds able to selectively block CB2 receptors may have therapeutic potential in controlling A beta-related pathology , due to their beneficial effects devoid of psychotropic consequences . Nicotinic alpha 7 receptors as a new target for treatment of cannabis abuse . Increasing use of cannabis makes the search for medications to reduce cannabis abuse extremely important . Here , we show that homomeric alpha7 nicotinic receptors are novel molecular entities that could be targeted in the development of new drugs for the treatment of cannabis dependence . In rats , systemic administration of the selective alpha7 nicotinic acetylcholine receptor antagonist methyllycaconitine ( MLA ) , but not the selective heteromeric non-alpha7 nicotinic acetylcholine receptor antagonist dihydrobetaerythroidine , ( 1 ) antagonized the discriminative effects of DB00470 ( THC ) , the main active ingredient in cannabis , ( 2 ) reduced intravenous self-administration of the synthetic cannabinoid P21554 receptor agonist WIN55,212-2 [ ( R ) -(+)-[2,3-dihydro-5-methyl-3[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-naphthalenyl)methanone , mesylate salt ] , and ( 3 ) decreased THC-induced dopamine elevations in the shell of the nucleus accumbens . Altogether , our results indicate that blockade of alpha7 nicotinic receptors reverses abuse-related behavioral and neurochemical effects of cannabinoids . Importantly , MLA reversed the effects of cannabinoids at doses that did not produce depressant or toxic effects , further pointing to alpha7 nicotinic antagonists as potentially useful agents in the treatment of cannabis abuse in humans . Opposite function of dopamine D1 and N-methyl-D-aspartate receptors in striatal cannabinoid-mediated signaling . It is well established that the cannabinoid and dopamine systems interact at various levels to regulate basal ganglia function . Although it is well known that acute administration of cannabinoids to mice can modify dopamine-dependent behaviors , the intraneuronal signaling pathways employed by these agents in the striatum are not well understood . Here we used knockout mouse models to examine the regulation of striatal extracellular-signal-regulated kinases 1 and 2 ( P27361 /2 ) signaling by behaviorally relevant doses of cannabinoids . This cellular pathway has been implicated as a central mediator of drug reward and synaptic plasticity . In C57BL/6J mice , acute administration of the cannabinoid agonists , (-)-11-hydroxydimethylheptyl-Δ8-tetrahydrocannabinol ( HU-210 ) and DB00470 ( Δ(9) -THC ) , promoted a dose- and time-dependent decrease in the phosphorylation of P27361 /2 in dorsal striatum . Co-administration of the P21554 cannabinoid receptor antagonist N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide(AM251) with HU-210 prevented P27361 /2 inactivation , indicating a requirement for activation of this receptor . In dopamine D1 receptor knockout animals treated with HU-210 , the magnitude of the HU-210-dependent decrease in striatal P27361 /2 signaling was greater than in wild-type controls . In contrast , HU-210 administration to N-methyl-D-aspartate receptor knockdown mice was ineffective at promoting striatal P27361 /2 inactivation . Genetic deletion of other potential P27361 /2 mediators , the dopamine D2 receptors or β-arrestin-1 or -2 , did not affect the HU-210-induced modulation of P27361 /2 signaling in the striatum . These results support the hypothesis that dopamine D1 receptors and N-methyl-D-aspartate receptors act in an opposite manner to regulate striatal P21554 cannabinoid receptor signal transduction . A prospective case-control study analyzes 12 thrombophilic gene mutations in Turkish couples with recurrent pregnancy loss . PROBLEM : Recurrent pregnancy loss ( RPL ) is a heterogeneous disorder . The contribution of specific thrombophilic genes to the pathophysiology of RPL has remained controversial . We evaluated the prevalences of 12 thrombophilic gene mutations among homogenous Caucasian couples with RPL and fertiles . METHOD : of study This was a prospective case-control study evaluating 272 women with RPL and 152 of their male partners , and a control group of 56 fertile couples . We investigated mutations including FV Leiden , factor V H1299R , factor II prothrombin G20210A , F XIII V34L , beta-fibrinogen -455G > A , plasminogen activator inhibitor-1 , P05106 L33P ( Q9Y251 -1 a/b L33P ) , P42898 C677T , P42898 A1298C , P12821 I/D , Apo B R3500Q , and Apo E . RESULTS : Overall , heterozygous mutations of FV Leiden , FXIII V34L , P05106 L33P , Apo E4 , and prothrombin G20210A and homozygous mutations of P05121 -1and P42898 C677T were associated with RPL . There was no meaningful association between RPL and other studied genes . CONCLUSION : In contrast to the other mutations and polymorphisms , FV Leiden , FXIII V34L , P05106 L33P , Apo E , prothrombin G20210A , P05121 and P42898 C677T gene mutations may help to identify the couples at risk for recurrent pregnancy loss . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . [ Cannabis use disorder and treatment of dependence ] . Cannabis , known as marijuana , has been used illicit drug by young people in the world . In our country , the number of user for cannabis is recently increased gradually . It has been suggested that regular use of cannabis might induce several adverse effects such as dependence syndrome , because DB00470 (THC) , a primary psychoactive component of cannabis , stimulates brain-reward areas through the activation of cannabinoid( P21554 ) receptor and induce drug-seeking behavior . Therefore , it is necessary to investigate and establish the medications for cannabis dependence . In fact , controlled laboratory studies and small open-label clinical studies have shown that several candidates of medications for cannabinoid dependence are identified . Further investigation in controlled clinical trials may produce the therapeutic benefit for treatment about cannabis-related problems . DB00470 -induced neurotoxicity depends on P21554 receptor-mediated c-Jun N-terminal kinase activation in cultured cortical neurons . Delta9- DB00470 ( THC ) , the main psychoactive ingredient of marijuana , induces apoptosis in cultured cortical neurons . THC exerts its apoptotic effects in cortical neurons by binding to the P21554 cannabinoid receptor . The P21554 receptor has been shown to couple to the stress-activated protein kinase , c-Jun N-terminal kinase ( JNK ) . However , the involvement of specific JNK isoforms in the neurotoxic properties of THC remains to be established . The present study involved treatment of rat cultured cortical neurons with THC ( 0.005-50 microM ) , and combinations of THC with the P21554 receptor antagonist , AM 251 ( 10 microM ) and pertussis toxin ( PTX ; 200 ng ml-1 ) . Antisense oligonucleotides ( AS ) were used to deplete neurons of P45983 and P45984 in order to elucidate their respective roles in THC signalling . Here we report that THC induces the activation of JNK via the P21554 receptor and its associated G-protein , Gi/o . Treatment of cultured cortical neurons with THC resulted in a differential timeframe of activation of the P45983 and P45984 isoforms . Use of specific P45983 and P45984 AS identified activation of caspase-3 and DNA fragmentation as downstream consequences of P45983 and P45984 activation . The results from this study demonstrate that activation of the P21554 receptor induces JNK and caspase-3 activation , an increase in Bax expression and DNA fragmentation . The data demonstrate that the activation of both P45983 and P45984 isoforms is central to the THC-induced activation of the apoptotic pathway in cortical neurons . Self-administration of cannabinoids by experimental animals and human marijuana smokers . Drug self-administration behavior has been one of the most direct and productive approaches for studying the reinforcing effects of psychoactive drugs , which are critical in determining their abuse potential . Cannabinoids , which are usually abused by humans in the form of marijuana , have become the most frequently abused illicit class of drugs in the United States . The early elucidation of the structure and stereochemistry of DB00470 ( THC ) in 1964 , which is now recognized as the principal psychoactive ingredient in marijuana , activated cannabinoid research worldwide . This review examines advances in research on cannabinoid self-administration behavior by humans and laboratory animals . There have been numerous laboratory demonstrations of the reinforcing effects of cannabinoids in human subjects , but reliable self-administration of cannabinoids by laboratory animals has only recently been demonstrated . It has now been shown that strong and persistent self-administration behavior can be maintained in experimentally and drug-naïve squirrel monkeys by doses of THC comparable to those in marijuana smoke inhaled by humans . Furthermore , reinforcing effects of some synthetic P21554 cannabinoid agonists have been recently reported using intravenous and intracerebroventricular self-administration procedures in rats and mice . These findings support previous conclusions that THC has a pronounced abuse liability comparable to other drugs of abuse under certain experimental conditions . Self-administration of THC by squirrel monkeys provides the most reliable animal model for human marijuana abuse available to date . This animal model now makes it possible to study the relative abuse liability of other natural and synthetic cannabinoids and to preclinically assess new therapeutic strategies for the treatment or prevention of marijuana abuse in humans . Inhibition of THC-induced effects on the central nervous system and heart rate by a novel P21554 receptor antagonist AVE1625 . P21554 antagonists such as AVE1625 are potentially useful in the treatment of obesity , smoking cessation and cognitive impairment . Proof of pharmacological action of AVE1625 in the brain can be given by antagonising the effects of DB00470 ( THC ) , a P21554 /CB2 agonist . Inhibition of THC-induced effects by AVE1625 was observed on Visual Analogue Scales ' alertness ' , ' feeling high ' , ' external perception ' , ' body sway ' and ' heart rate ' . Even the lowest dose of AVE1625 20 mg inhibited most of THC-induced effects . AVE1625 did not have any effect on psychological and behavioural parameters or heart rate by itself . After THC and AVE1625 administration , changes on electroencephalography were observed . This study shows a useful method for studying the effects of P21554 antagonists . AVE1625 penetrates the brain and antagonises THC-induced effects with doses at or above 20 mg . The stimulation of ketogenesis by cannabinoids in cultured astrocytes defines carnitine palmitoyltransferase I as a new ceramide-activated enzyme . The effects of cannabinoids on ketogenesis in primary cultures of rat astrocytes were studied . Delta9- DB00470 ( THC ) , the major active component of marijuana , produced a malonyl- DB01992 -independent stimulation of carnitine palmitoyltransferase I ( CPT-I ) and ketogenesis from [14C]palmitate . The THC-induced stimulation of ketogenesis was mimicked by the synthetic cannabinoid HU-210 and was prevented by pertussis toxin and the P21554 cannabinoid receptor antagonist SR141716 . Experiments performed with different cellular modulators indicated that the THC-induced stimulation of ketogenesis was independent of cyclic AMP , Ca2+ , protein kinase C , and mitogen-activated protein kinase ( MAPK ) . The possible involvement of ceramide in the activation of ketogenesis by cannabinoids was subsequently studied . THC produced a P21554 receptor-dependent stimulation of sphingomyelin breakdown that was concomitant to an elevation of intracellular ceramide levels . Addition of exogenous sphingomyelinase to the astrocyte culture medium led to a MAPK-independent activation of ketogenesis that was quantitatively similar and not additive to that exerted by THC . Furthermore , ceramide activated CPT-I in astrocyte mitochondria . Results thus indicate that cannabinoids stimulate ketogenesis in astrocytes by a mechanism that may rely on P21554 receptor activation , sphingomyelin hydrolysis , and ceramide-mediated activation of CPT-I . Effects of delta9-THC on P01282 -induced prolactin secretion in anterior pituitary cultures : evidence for the presence of functional cannabinoid P21554 receptors in pituitary cells . Peripheral administration of cannabinoid P21554 receptor agonists to laboratory rats induce a brief rise in plasma prolactin ( PRL ) levels followed by a prolonged decrease in PRL secretion from the pituitary . While the inhibitory component of this biphasic response depends on the cannabinoid-induced activation of dopamine release from hypothalamic terminals located in the median eminence , the neurobiological mechanisms underlying the activation phase of PRL release remains to be explained . In the present study the possible direct effect of the cannabinoid receptor agonist DB00470 ( THC ) on prolactin secretion and DB02527 accumulation was examined in anterior pituitary cultures . THC ( 0.1 and 1 microM ) increased DB02527 levels , and induced PRL release ( 1 and 10 mu ) . THC did not affect vasoactive intestinal peptide ( P01282 , 0.5 microM ) induced DB02527 accumulation in pituitary cultures , showing additive effects at THC 1 microM concentration . However , THC did prevent P01282 -dependent increases in prolactin secretion . These results indicate that THC , through a direct pituitary action , activates both the synthesis of DB02527 and PRL release and interferes with intracellular mechanisms involved in PRL secretion by P01282 . These actions could be mediated through cannabinoid P21554 receptors which were found to be present in anterior pituitary cells , including lactotrophs , as revealed by immunocytochemistry with a specific polyclonal antibody raised against the P21554 receptor protein . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . [ The effect of bunazosin vs captopril on hemodynamic and neurohumoral parameters in patients with congestive heart failure ] . The hemodynamic parameters ( right atrial pressure , mean pulmonary artery pressure , pulmonary capillary wedge pressure , cardiac index , heart rate , blood pressure ) and neurohumoral responses ( alpha- P01160 , plasma renin activity , aldosterone , angiotensin II ) of DB01197 , oral P12821 inhibitor , and Bunazosin , oral alpha 1-blocker , were investigated in 28 patients with congestive heart failure at rest and after exercise . These data were analysed in both acute and chronic phases . 1 ) Acute effect . DB01197 produced significant improvement of neurohumoral factors at rest and also after exercise . Bunazosin reduced alpha- P01160 , but other neurohumoral factors did not change . Bunazosin produced significant hemodynamic improvement both at rest and after exercise . 2 ) Chronic effect . DB01197 produced significant hemodynamic improvement both at rest and after exercise . Improvement of neurohumoral factors in acute phase was also preserved at chronic phase . On Bunazosin , improvement of hemodynamics at acute phase was also preserved at chronic phase without deterioration of neurohumoral factors . Differential effects of delta9-tetrahydrocannabinol and methanandamide in P21554 knockout and wild-type mice . Mice devoid of P21554 cannabinoid receptors ( P21554 -/- mice ) provide a unique opportunity to further investigate the role of P21554 receptors in exocannabinoid and endocannabinoid effects . P21554 -/- mice ( N = 18 ) and their wild-type littermates ( P21554 +/+ mice ; N = 12 ) were placed in standard mouse operant chambers and trained to lever press under a fixed ratio 10 schedule of reinforcement . When stable lever press responding under the fixed ratio 10 schedule had been established , cannabinoids and noncannabinoids were administered to both groups . P21554 +/+ mice acquired the lever press response more readily than P21554 -/- mice . Delta(9)- DB00470 ( Delta(9)-THC ) decreased lever press responding in P21554 +/+ mice only , whereas methanandamide , a metabolically stable endocannabinoid analog , produced similar response rate decreases in both genotypic groups . Similar to Delta(9)-THC , another endocannabinoid analog , ( R ) - ( 20-cyano-16,16-dimethyl docosa-cis-5,8,11,14-tetraeno ) -1'-hydroxy-2'-propylamine ( O-1812 ) , decreased responding in P21554 +/+ mice , but not in P21554 -/- mice . The P21554 receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1- ( 2,4-dichlorophenyl-4-methyl-1H-pyrazole-3-carboxamide hydrochloride ( SR141716A ) blocked the effects of Delta(9)-THC , but not those of methanandamide . Because methanandamide binds poorly to CB2 receptors , these results suggest possible non- P21554 , non-CB2 mechanisms of action for methanandamide-induced behavioral disruption of lever press responding . DB00898 and morphine elicited greater response decreases in P21554 -/- mice than in P21554 +/+ mice , suggesting a possible role of P21554 receptors in the rate disruptive effects of these drugs . In contrast , diazepam did not produce between group differences , suggesting that P21554 receptors are not involved in diazepam-induced disruption of lever press responding . Extended exposure to a palatable cafeteria diet alters gene expression in brain regions implicated in reward , and withdrawal from this diet alters gene expression in brain regions associated with stress . Like people , rodents exposed to energy-rich foods over-eat and become overweight . Removal of this diet activates stress systems , which may explain why people have difficulty dieting . We exposed rats to energy-rich foods in order to identify changes in the brain induced by that diet and by its removal . Sprague Dawley rats were fed lab-chow or an energy-rich cafeteria diet ( plus chow ) . Following 6 or 15 weeks , half of each group was switched to the opposing diet . Rats were culled 48-h later . We measured fat mass , plasma hormones , and assessed brains for mRNA expression of several genes . Cafeteria-fed rats consumed more kilojoules , weighed more and had elevated leptin ( plus reduced O00230 at 15 weeks ) relative to chow-fed rats . Fifteen weeks of cafeteria diet suppressed μ-opioid and P21554 receptor mRNA in the VTA , but elevated amygdala GR , and 6 weeks of cafeteria diet reduced P23560 , compared to chow-fed rats . Rats switched to the cafeteria diet ate similar amounts as rats maintained on the diet , and switching to cafeteria diet after 15 weeks reduced amygdala GR expression . Rats switched to chow ate less than rats maintained on chow , and switching to chow following 15 weeks of cafeteria diet increased hypothalamic P06850 mRNA . Therefore , 15 weeks of cafeteria diet produced changes in brain regions implicated in reward processes . Switching these rats to chow activated the Q9Y251 axis , while switching chow-fed rats to the cafeteria diet decreased GR expression in the amygdala , a region associated with stress . These findings have implications for dieting in humans . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Attenuation of inducible nitric oxide synthase gene expression by delta 9-tetrahydrocannabinol is mediated through the inhibition of nuclear factor- kappa B/Rel activation. delta 9- DB00470 ( delta 9-THC ) a prototypic compound belonging to the family of agents known as cannabinoids , produces a wide variety of biological effects , including inhibition of immune function . The putative mechanism for cannabinoid biological action involves binding to cannabinoid receptor types 1 and 2 ( P21554 and CB2 ) to negatively regulate adenylate cyclase and inhibit intracellular signaling via the DB02527 cascade . In the current study , we show that delta 9-THC produces a marked inhibition of inducible nitric oxide synthase ( P35228 ) transcription and nitric oxide production by the macrophage line RAW 264.7 in response to lipopolysaccharide ( LPS ) . Analysis of RAW 264.7 cell RNA demonstrated transcripts for CB2 but not P21554 . Treatment of RAW 264.7 with delta 9-THC inhibited forskolin-stimulated DB02527 production in a dose-related manner , verifying the expression of functional cannabinoid receptors by this cell line . P35228 transcription , which is regulated in part by the nuclear factor-kappa B/Rel ( NF-kappa B/Rel ) family of transcription factors , has been shown to be under the control of the DB02527 signaling cascade . We demonstrate that delta 9-THC inhibits the activation and binding of NF-kappa B/Rel proteins to their cognate DNA site , kappa B , in response to LPS stimulation . LPS treatment of RAW 264.7 cells also induced the activation of the DB02527 cascade , as indicated by an increase in binding of nuclear factors to the DB02527 response element . Activation of CRE binding proteins was inhibited by delta 9-THC . DB02587 treatment of RAW 264.7 cells induced both kappa B and DB02527 response element binding activity and was likewise inhibited by delta 9-THC . Collectively , this series of experiments indicates that NF-kappa B/Rel is positively regulated by the DB02527 cascade to help initiate P35228 gene expression in response to LPS stimulation of macrophages . This activation of P35228 is attenuated by delta 9-THC through the inhibition of DB02527 signaling . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . Identification of a high-affinity binding site involved in the transport of endocannabinoids . Phytocannabinoids , such as the principal bioactive component of marijuana , delta9-tetrahydrocannabinol , have been used for thousands of years for medical and recreational purposes . DB00470 and endogenous cannabinoids ( e.g. , anandamide ) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors ( P21554 and CB2 ) . The biosynthesis and physiology of anandamide is well understood , but its mechanism of uptake ( resulting in signal termination by fatty acid amide hydrolase ) has been elusive . Mounting evidence points to the existence of a specific anandamide transport protein ; however , no direct evidence for this protein has been provided . Here , we use a potent , competitive small molecule inhibitor of anandamide uptake ( LY2318912 , IC50 7.27 +/- 0.510 nM ) to identify a high-affinity , saturable anandamide transporter binding site ( LY2318912 ; K(d) = 7.62 +/- 1.18 nM , B(max) = 31.6 +/- 1.80 fmol/mg protein ) that is distinct from fatty acid amide hydrolase . Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function . Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling , and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 .

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interacts_with DB01268?

multiple_choice

A new gene mapping resource : interspecies hybrids between Père David 's deer ( Elaphurus davidianus ) and red deer ( Cervus elaphus ) . Three male F1 hybrids between Père David 's deer and red deer were mated to red deer to produce 143 backcross calves . The pedigrees are a rare example of a fertile hybrid between evolutionarily divergent species . We examined the use of these families for genetic mapping of evolutionarily conserved ( Type I ) loci by testing for genetic linkage between five species-specific protein variants and 12 conserved DNA probes . Two probes were homologous , and the remainder syntenic , to the protein coding loci in cattle or humans . Using six restriction enzymes , each DNA probe detected one or more restriction fragments specific to Père David 's deer . Linkage analyses among the species-specific variants placed the loci into four linkage groups within which linkage between adjacent loci and gene order was supported by a LOD > 3 . The linkage groups were ( P02790 , P68871 ) - P01225 - P11117 , P00338 - P06127 - P01344 , P12645 - ( GC , ALB ) - ( P10721 , P16234 ) and P01130 - P01024 - P05230 . Southern and protein analysis of P00338 and ALB provided identical segregation data . These linkage groups were consistent with the cattle gene map and provide new information for comparing the gene maps of ruminants , humans and mice . The deer hybrids are an important new resource that can contribute to the comparative analysis of the mammalian genome . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . An online coupled cell membrane chromatography with LC/MS method for screening compounds from Aconitum carmichaeli Debx. acting on P35968 . An online analytical method coupling high expression vascular endothelial growth factor receptor ( VEGFR ) cell membrane chromatography ( VEGFR-CMC ) with high performance liquid chromatography mass spectrometry ( LC/MS ) for screening and identification of active component from traditional Chinese herb Aconitum carmichaeli Debx. acting on P35968 was established . Through a 10-port column switcher , factions separated by VEGFR-CMC column ( first dimension ) were transferred and were adsorbed on an enrichment column . Then , these fractions were sent into LC/MS system ( second dimension ) immediately and directly for separation and preliminary identification , respectively . DB01268 malate ( SN ) was used as positive control , while nifedipine ( NF ) , dexamethasone acetate ( DX ) , methoxyamine hydrochloride ( MT ) and atenolol ( AT ) as negative controls . The specification of this VEGFR-CMC-online-LC/MS method was validated by competitive displacement test . As a result , mesaconitine ( O60682 ) , aconitine ( AC ) , and hypaconitine ( HPC ) were identified as the active constituents acting on P35968 . The in vitro inhibition activity of starting extract of Aconitum carmichaeli Debx. , O60682 , AC , and HPC on HEK293/VEGFR cell viability by MTT test , separately . The in vitro inhibition activity of O60682 , AC , and HPC on vascular endothelial growth factor ( P15692 ) secretion of HEK293/VEGFR cell was tested by P15692 -ELISA assay . The screening results given by the system offered additional exemplification supporting this online coupling method and gave new evidence to the development of anti-tumor drug from natural products . Gastrointestinal stromal tumor . Gastrointestinal stromal tumors ( GISTs ) are the most common mesenchymal tumors of the gastrointestinal tract . These form a distinct category of tumors characterized by oncogenic mutations of the P10721 receptor tyrosine kinase in a majority of patients . P10721 is used not only for diagnosis but also for targeted therapy of GISTs . Imatinib , a tyrosine kinase inhibitor , is widely used in the treatment of advanced and metastatic GISTs and has been recently employed in the neo adjuvant and adjuvant set-up with encouraging results . Certain specific mutations in an exon ( such as in exon 9 ) of the P10721 gene result in GISTs that are relatively unresponsive to the Imatinib treatment . New therapeutic agents like DB01268 have now been approved for the treatment of Imatinib-resistant GIST . This review summarizes the salient features of GIST along with a detailed review of targeted multi-disciplinary approach to the treatment of these special tumors . DB01268 . DB01268 is an oral multikinase inhibitor that blocks the vascular endothelial growth factor receptor ( VEGFR ) , platelet-derived growth factor receptor ( P09619 ) alpha and beta , c-kit , and other receptors . These attributes have proven to be efficacious in the treatment of metastatic renal cell carcinoma ( RCC ) and unresectable gastrointestinal stromal tumors ( GIST ) . Most side effects , including hypertension , hand-foot syndrome , and diarrhea are generally well manageable . Clinical trials are underway to determine the efficacy of sunitinib in other tumor types including metastatic breast , colorectal , and lung cancers . This chapter will detail the preclinical data leading to the results of the pivotal phase III clinical trials that have led to the widespread use of sunitinib in metastatic RCC and advanced GIST . Nonclinical safety evaluation of sunitinib : a potent inhibitor of P15692 , PDGF , P10721 , P36888 , and P07949 receptors . DB01268 malate ( SUTENT ) is a multitargeted receptor tyrosine kinase ( RTK ) inhibitor that is approved multinationally for the treatment of imatinib-resistant/-intolerant gastrointestinal stromal tumor and advanced renal cell carcinoma . This paper characterizes the organ toxicity of sunitinib in Sprague-Dawley rats and cynomolgus monkeys , and the reversibility of any treatment-induced effects . Rats and monkeys received sunitinib ( 0-15 and 0-20 mg/kg/day , respectively ) orally on a consecutive daily dosing schedule for thirteen weeks or on an intermittent daily dosing schedule for up to nine months . Clinical observations and laboratory parameters were recorded . Necropsy was conducted following treatment/recovery periods , and histologic examinations were performed . In rats , sunitinib was generally tolerated at 0.3 and 1.5 mg/kg/day , and findings were reversible . In monkeys , the level at which there were no observed adverse effects was 1.5 mg/kg/day , and findings were similarly reversible ( except for uterine/ovarian weight changes and skin pallor ) . Data suggest that inhibition of multiple RTK pathways may induce pharmacologic effects on organ systems in nonclinical species . Key pharmacologic effects of sunitinib included reversible inhibition of neovascularization into the epiphyseal growth plate , and impaired corpora lutea formation and uterine development during estrus . Similar observations have been noted with this class of RTK signaling inhibitors and are consistent with pharmacologic perturbations of physiologic/angiogenic processes associated with the intended molecular targets . The effects of P04035 inhibitor on vascular progenitor cells . Circulating bone marrow-derived vascular progenitor cells contribute to angiogenesis , atherosclerosis , and the response to vascular injury . These vascular progenitor cells consist of two cell groups , endothelial progenitor cells ( EPCs ) and smooth muscle progenitor cells ( SMPCs ) . Although P04035 inhibitors ( statins ) have been reported to inhibit atherosclerosis partially by increased EPCs , the effects of statins on SMPCs are unclear . Therefore , we investigated the relationship between EPCs and SMPCs and whether pravastatin has atheroprotective effects on SMPCs . Peripheral mononuclear cells ( MNCs ) were isolated and cultured on fibronectin-coated dishes in SMPC medium . MNCs were stained with acetylated low density lipoprotein and lectin , or alpha-smooth muscle actin , and cell numbers were counted . mRNA expression and vascular endothelial growth factor ( P15692 ) protein synthesis of MNCs were evaluated . DB00175 significantly increased the number of EPC and decreased the number of SMPC. mRNA expression of P15692 , endothelial nitric oxide synthase , P15692 receptor-2 ( P35968 ) , and Akt were up-regulated , and P15692 secretion was increased by pravastatin . The present study demonstrated that pravastatin has promotive effects on the differentiation from MNCs to EPC cells , while inhibitory effects to SMPC cells . Our findings suggest a previously unreported mechanism of the effect of statin therapy on vascular progenitor cells . DB01268 in metastatic thymic carcinomas : laboratory findings and initial clinical experience . BACKGROUND : Thymic carcinoma ( TC ) is a rare aggressive tumour . Median survival with current treatments is only 2 years . DB01268 is a multi-targeted tyrosine kinase inhibitor that has shown benefit in various other cancers . METHODS : Laboratory analyses of snap-frozen tumour tissues were performed to detect activation and genetic mutations of receptor tyrosine kinases ( RTKs ) in TC samples . On the basis of molecular analyses showing activation of multiple RTKs in their tumour , four patients with metastatic TCs refractory to conventional therapies were treated with sunitinib according to standard protocols . RESULTS : RTK analysis in three of the patients showed activation of multiple RTKs , including platelet-derived growth factor-beta and vascular endothelial growth factor 3 . Mutations of P00533 , c- P10721 , P01116 , and P15056 genes were not found . Administration of sunitinib yielded a partial remission ( lasting 2 to 18+ months ) according to the RECIST criteria in three patients and stable disease with excellent metabolic response in DB09150 -PET in another one . The overall survival with sunitinib treatment ranges from 4 to 40+ months . Withdrawal of the drug in one patient prompted rapid tumour progression that could be controlled by re-administration of sunitinib . CONCLUSIONS : DB01268 is an active treatment for metastatic TC . A panel of molecular analyses may be warranted for optimal patient selection . Serotonin transporter interacts with the PDGFβ receptor in DB00102 -induced signaling and mitogenesis in pulmonary artery smooth muscle cells . The serotonin transporter ( P31645 ) and the platelet-derived growth factor receptor ( P09619 ) have been implicated in both clinical and experimental pulmonary hypertension ( PH ) and the facilitation of pulmonary artery smooth muscle cell ( PASMC ) growth . To gain a better understanding of the possible relationship of these two cell surface molecules we have explored interactions between P31645 and P09619 . We have previously demonstrated that P31645 transactivates PDGFRβ in serotonin-stimulated PASMC proliferation . We now provide evidence for a role for P31645 in DB00102 signaling and PASMC proliferation by using pharmacological inhibitors , genetic ablation , and construct overexpression of P31645 . The results show that four tested P31645 blockers dose dependently inhibit PDGF-stimulated human and bovine PASMC proliferation with comparable efficacy to that of P09619 inhibitors , whereas P28222 or 5- Q13049 receptor inhibitors had no effect . Combinations of the P31645 and P09619 inhibitors led to synergistic/additive inhibition . Similarly , PDGF-induced PASMC proliferation was attenuated by small interfering RNA downregulation of P31645 . Inhibition of P31645 in PASMCs attenuated PDGF-induced phosphorylation of PDGFRβ , Akt , and p38 but not Erk . Overexpression of P31645 in HEK293 cells led to enhanced Akt phosphorylation by PDGF , which was blunted by a P31645 PDZ motif mutant , indicating the mechanistic need for the PDZ motif of P31645 in PDGF signaling . Furthermore , coimmunoprecipitation experiments showed that P31645 and PDGFRβ become physically associated upon PDGF stimulation . In total , the data show for the first time an important interactive relationship between P31645 and the PDGFRβ in the production of PASMC proliferation triggered by PDGF that may be important in PH . Novel approaches to gastrointestinal stromal tumors resistant to imatinib and sunitinib . Gastrointestinal stromal tumors ( GIST ) are rare tumors of mesenchymal origin that may arise anywhere along the gastrointestinal tract or in the peritoneum . In most cases , GIST harbor mutations of P10721 or P16234 . Imatinib mesylate ( IM ) , a small-molecule tyrosine kinase inhibitor developed for the treatment of chronic myeloid leukemia , has been shown to be active against these mutations and has significant activity in patients with metastatic GIST . However , resistance to IM emerges after a median of 24 months of treatment . DB01268 malate ( SU ) has been approved for the treatment of patients with IM-resistant advanced GIST , but the median progression-free survival in this setting is only 6 months . This article reviews the current knowledge regarding IM and SU resistance in GIST , as well as the available options for the management of GIST resistant to IM and SU . DB01268 ( SUTENT , SU11248 ) suppresses tumor growth and induces apoptosis in xenograft models of human hepatocellular carcinoma . Hepatocellular carcinoma ( HCC ) is the fifth most common and third deadliest primary neoplasm . Since HCC is a particularly vascular solid tumor , we determined the antitumor and antiangiogenic activities of sunitinib malate , a potent inhibitor of two receptors involved in angiogenesis - vascular endothelial growth factor receptor ( VEGFR ) and platelet-derived growth factor receptor ( P09619 ) . In the present study , we reported that treatment of HepG2 and SK-Hep-1 cells with sunitinib led to growth inhibition and apoptosis in a dose-dependent fashion . DB01268 inhibited phosphorylation of P35968 at Tyr951 and P09619 at Tyr1021 both in vitro and in vivo . DB01268 also suppressed tumor growth of five patient-derived xenografts . DB01268 -induced tumor growth inhibition was associated with increased apoptosis , reduced microvessel density and inhibition of cell proliferation . This study provides a strong rationale for further clinical investigation of sunitinib in patients with hepatocellular carcinoma . Synthesis , in silico , in vitro , and in vivo investigation of 5-[¹¹C]methoxy-substituted sunitinib , a tyrosine kinase inhibitor of P35968 . DB01268 ( SU11248 ) is a highly potent tyrosine kinase inhibitor targeting vascular endothelial growth factor receptor ( VEGFR ) . Radiolabeled inhibitors of receptor tyrosine kinases ( RTKs ) might be useful tools for monitoring RTKs levels in tumor tissue giving valuable information for anti-angiogenic therapy . Herein we report the synthesis of 5-methoxy-sunitinib 5 and its (11)C-radiolabeled analog [ (11)C ] -5 . The non-radioactive reference compound 5 was prepared by Knoevenagel condensation of 5-methoxy-2-oxindole with the corresponding substituted 5-formyl-1H-pyrrole . A binding constant ( K(d) ) of 20 nM for 5 was determined by competition binding assay against P35968 . In addition , the binding mode of sunitinib and its 5-methoxy substituted derivative was studied by flexible docking simulations . These studies revealed that the substitution of the fluorine at position 5 of the oxindole scaffold by a methoxy group did not affect the inhibitor orientation , but affected the electrostatic and van der Waals interactions of the ligand with residues near the DFG motif of P35968 . 5-[(11)C]methoxy-sunitinib ( [ (11)C ] -5 ) was synthesized by reaction of the desmethyl precursor with [(11)C]CH(3)I in the presence of DMF and NaOH in 17 ± 3 % decay-corrected radiochemical yield at a specific activity of 162-205 GBq/μmol ( EOS ) . In vivo stability studies of [ (11)C ] -5 in rat blood showed that more than 70 % of the injected compound was in blood stream , 60 min after administration . Initial interrogation , confirmation and fine mapping of modifying genes : P40763 , P01584 and P15260 determine cystic fibrosis disease manifestation . We have used a stepwise approach consisting of initial interrogation , confirmation and fine mapping to analyze P40763 , P01584 and P15260 as modifiers of cystic fibrosis disease building upon the data and sample collection of the European Cystic Fibrosis Twin and Sibling Study . We have observed direct correlation between the length of the intronic microsatellite STAT3Sat to P40763 expression levels among F508del- P13569 homozygous patients ( P=0.0075 ) , and an association of longer STAT3Sat-alleles with the presence of P13569 -mediated residual chloride secretion ( P=0.0031 ) , measured as the manifestation of the CF basic defect in intestinal tissue . Both , family-based analysis by P04053 and case-reference comparison identified consistently the same intragenic P01584 haplotype as a risk variant ( P(raw)=0.055 for P04053 , P(raw) < 0.3 for case-reference comparison ) . Using haplotype-guided hierarchical fine mapping , we have identified two single nucleotide exchanges for which concordant and discordant sibling pairs differ at a 7 kb-spanning core haplotype in P15260 ( P(raw)=0.0113 ) . Taken together , our findings imply that immunorelevant pathways and ion secretion , dominated by P13569 in intestinal and respiratory epithelium , merge at the level of the epithelial cell to integrate the signaling of cytokines due to innate and acquired immune defense . Polymorphisms of dopamine receptor/transporter genes and risk of non-small cell lung cancer . BACKGROUND : The dopaminergic pathway may be of interest in assessing risk of non-small cell lung cancer ( NSCLC ) . Dopamine receptors are expressed in alveolar epithelial cells and human lung tumours , and dopamine inhibits both cell proliferation in vitro and growth of lung tumour xenografts in nude mice . Moreover , dopamine selectively inhibits the vascular permeability and angiogenic activity of vascular endothelial growth factor ( P15692 / P15692 ) . The bioavailability of dopamine is regulated by dopamine receptors D2 ( P14416 ) , D4 ( P21917 ) and dopamine transporter 1 ( Q01959 / Q01959 ) genes . METHODS : We have analysed 10 single nucleotide polymorphisms in P14416 , P21917 and Q01959 / Q01959 genes in relation to lung cancer risk in a case-control study of smoking subjects . The study subjects were 413 healthy individuals from general population and 335 NSCLC cases . Both cases and controls were Caucasians of Norwegian origin . RESULTS : We demonstrate that P14416 polymorphisms -141Cdel , 3208G > T , TaqIB ; P21917 -521C > T and Q01959 / Q01959 -1476T > G are associated with a two- to five-fold increased NSCLC risk . The variant alleles of P14416 1412A > G and 960C > G had protective effects . CONCLUSION : The dopamine receptor/transport gene polymorphisms are associated with the risk of NSCLC among smokers . The data show that the polymorphisms resulting in lower dopamine bioavailability were associated with increased risk of NSCLC . DB01268 enhances antitumor effects against chemotherapy-resistant bladder cancer through suppression of P27361 /2 phosphorylation . Bladder cancer patients who are refractory to chemotherapy have a poor prognosis . Furthermore , additional chemotherapies provide little benefit to patients who have relapsed after an initial response . Recently , it was reported that several molecular pathways are implicated in bladder carcinogenesis , including the epidermal growth factor receptor ( P00533 ) pathway , the vascular endothelial growth factor ( P15692 ) pathway and the Ras-MAPK pathway . We hypothesized that sunitinib would be effective in bladder cancer as it is an oral inhibitor of multiple receptor tyrosine kinases , including P15692 receptors , platelet derived growth factor ( PDGF ) receptors and stem cell factor receptor ( c- P10721 ) , and is a standard first-line treatment of advanced clear cell renal carcinoma . In the present study , the antiproliferative effects of sunitinib were clearly demonstrated in KK47 , KK47/DDP20 and KK47/ADR cell lines in vitro due to the suppression of P27361 /2 phosphorylation . In a mouse model , the antitumor effects of sunitinib were again clearly seen . Also , treatment with sunitinib decreased the abundance of regulatory T cells ( Tregs ) . However , cytotoxic T lymphocyte ( CTL ) activity was not induced sufficiently as compared with an IFN-α-treated group . Our results suggested that sunitinib was effective in chemotherapy-resistant bladder cancer patients . On the other hand , these findings provided the rationale for combination therapy with sunitinib and immune-based cancer therapy for advanced malignancies to prevent the occurrence of rebound phenomena . Tyrosine kinase inhibition in renal cell carcinoma and gastrointestinal stromal tumours : case reports . BACKGROUND : DB01268 malate is approved multinationally for the treatment of metastatic renal cell carcinoma ( mRCC ) and advanced imatinib-refractory gastrointestinal stromal tumour ( GIST ) . Greater exposure to sunitinib is associated with improved efficacy . Therefore , minimising the impact of adverse events ( AEs ) on patient quality of life is important to enable patients to achieve optimal exposure to sunitinib and maximum clinical benefit . DESIGN : This report describes four patient cases in which sunitinib was utilised for the management of advanced malignancies : two cases describe mRCC patients who received first-line sunitinib and two cases describe the use of targeted therapies , including sunitinib , in patients with advanced GIST . RESULTS : In all four cases , effective AE management enabled patients to receive long-term therapy with sunitinib and achieve sustained clinical benefit . The two mRCC cases show prolonged responses and manageable AEs with sunitinib . The two GIST cases demonstrate that patients with imatinib-refractory GIST with P10721 exon 9 mutations , including elderly patients , can achieve sustained responses to sunitinib . CONCLUSIONS : These case studies support the long-term efficacy and safety of sunitinib in the management of mRCC and imatinib-refractory GIST and demonstrate how AE management can be used to optimise patient responses . Identification of microRNAs involved in the modulation of pro-angiogenic factors in atherosclerosis by a polyphenol-rich extract from propolis . New vessel formation plays a critical role in the progression and vulnerability of atherosclerotic lesions . It has been shown that polyphenols from propolis attenuate the progression of atherosclerosis and also exert inhibitory effects on angiogenic factors . However , the mechanisms underlying these effects are not completely understood . Thus , this study aimed to identify microRNAs ( miRNAs ) involved in the modulation of pro-angiogenic factors in the atherosclerotic plaques of P01130 gene knockout mice treated with a polyphenol-rich extract of Chilean propolis . The progression of the atherosclerotic lesions was significantly attenuated in treated mice compared with control mice . Using microarray analysis and a bioinformatic approach , we identified 29 differentially expressed miRNAs . Many of these miRNAs were involved in biological processes associated with angiogenesis , such as the cell cycle , cell migration , cell growth and proliferation . Among them , three miRNAs ( miR-181a , miR-106a and miR-20b ) were over-expressed and inversely related to the expression of Vegfa ( vascular endothelial growth factor A ) and Hif1a ( hypoxia inducible factor 1 alpha ) . In addition , P15692 protein expression was attenuated in histological sections obtained from the aortic sinuses of treated mice . P15692 is a key pro-angiogenic factor in atherosclerotic plaques , and Hif1a , which is expressed in the necrotic nucleus of the atheroma , is its main inducer . We found a correlation between the over-expression of miR-181a , miR-106a and miR-20b and their target genes , Hif1a and Vegfa , which is consistent with attenuation of the atherosclerotic lesion . In conclusion , our data analysis provides evidence that the anti-angiogenic effects of polyphenols from Chilean propolis can be modulated by miRNAs , in particular miR-181a , miR-106a and miR-20b . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Expression of vascular endothelial growth factor ( P15692 ) and its receptors P17948 and P35968 in primary and recurrent WHO grade III meningiomas . AIMS : WHO grade III meningiomas are malignant neoplasms for which new and more targeted treatment strategies are urgently needed . Although clinical trials investigating anti-angiogenic vascular endothelial growth factor ( P15692 ) targeted therapies are currently recruiting , knowledge about the expression of P15692 and P15692 receptors remains to be determined . METHODS : We investigated the expression of P15692 and its receptors P17948 and P35968 in 32 WHO grade III meningioma samples by immunohistochemistry . Furthermore , we performed in-situ hybridisation for P15692 . RESULTS : We found low P15692 expression in tumor and endothelial cells . Highest P15692 expression levels were seen in peri-necrotic tumor cells potentially suffering from hypoxia . P17948 and 2 were virtually absent on tumor cells , although endothelial cells displayed significantly higher levels reaching stronger expression for P35968 than P17948 . CONCLUSIONS : Our findings showing constant expression levels of P35968 in endothelial cells serve as a first indication that the use of small tyrosine kinase inhibitors such as DB01268 directly targeting the P15692 -receptors might be worth testing , also in the clinical context in cases of therapy-refractory meningiomas . Further investigations are needed to study the response to drugs targeting the P15692 pathway in relation to the expression profile of P15692 and its receptors in high grade meningiomas . Effects of preset sequential administrations of sunitinib and everolimus on tumour differentiation in Caki-1 renal cell carcinoma . BACKGROUND : DB01268 ( VEGFR/ P09619 inhibitor ) and everolimus ( P42345 inhibitor ) are both approved for advanced renal cell carcinoma ( RCC ) as first-line and second-line therapy , respectively . In the clinics , sunitinib treatment is limited by the emergence of acquired resistance , leading to a switch to second-line treatment at progression , often based on everolimus . No data have been yet generated on programmed alternating sequential strategies combining alternative use of sunitinib and everolimus before progression . Such strategy is expected to delay the emergence of acquired resistance and improve tumour control . The aim of our study was to assess the changes in tumours induced by three different sequences administration of sunitinib and everolimus . METHODS : In human Caki-1 RCC xenograft model , sunitinib was alternated with everolimus every week , every 2 weeks , or every 3 weeks . Effects on necrosis , hypoxia , angiogenesis , and EMT status were assessed by immunohisochemistry and immunofluorescence . RESULTS : DB01268 and everolimus programmed sequential regimens before progression yielded longer median time to tumour progression than sunitinib and everolimus monotherapies . In each group of treatment , tumour growth control was associated with inhibition of P42345 pathway and changes from a mesenchymal towards an epithelial phenotype , with a decrease in vimentin and an increase in P12830 expression . The sequential combinations of these two agents in a RCC mouse clinical trial induced antiangiogenic effects , leading to tumour necrosis . CONCLUSIONS : In summary , our study showed that alternate sequence of sunitinib and everolimus mitigated the development of mesenchymal phenotype compared with sunitinib as single agent . Expression and mutational status of treatment-relevant targets and key oncogenes in 123 malignant salivary gland tumours . BACKGROUND : Malignant tumours of the salivary glands ( MSGT ) are rare and pleomorphic entities . Patients with advanced disease may benefit from targeted therapy ; however , specific targets for optimising and personalising treatments are yet to be identified . DESIGN : Immunohistochemistry for C- P10721 , P00533 , P04626 , P15941 , phospho- P42345 , androgen/estrogens/progesterone receptors and Ki67 was carried out and evaluated in terms of progression-free and overall survival . High throughput molecular screening of key oncogenes was done in 107 patients using routine diagnostic methods and Sequenom technology . RESULTS : Several therapy leads were identified , including high levels of P04626 and androgen receptors in salivary duct carcinomas , C- P10721 in myoepithelial carcinomas and P00533 in mucoepidermoid carcinomas . Recurrent mutations involving downstream elements of the P00533 pathway were found in P01112 , notably in tumours with a myoepithelial component , and in other key oncogenes ( P01116 / P01111 /PI3KCA/ P15056 /MAP2K ) . On the other hand , < 1 % of samples had P00533 or P04626 mutations . CONCLUSION : Several tumour subtypes overexpressed targets of directed therapies suggesting potential therapy leads . Genotyping results suggest activation downstream of P00533 in 18 of the 107 samples that could be associated with low efficacy of P00533 inhibitors . Other molecules , such as PI3K/MEK or P42345 inhibitors , may have anti-tumour activity in this subgroup . The high mutation rate in P01112 highlights a novel key oncogenic event in MSGT . Regulation of gene expression in melanoma : new approaches for treatment . The molecular changes associated with the transition of melanoma cells from radial growth phase ( RGP ) to vertical growth phase ( VGP , metastatic phenotype ) are not yet well defined . We have demonstrated that the progression of human melanoma is associated with loss of expression of the transcription factor P05549 . In metastatic melanoma cells , this loss resulted in overexpression of P43121 / P43121 , P08253 , the thrombin receptor ( P25116 ) , and lack of c- P10721 expression . The transition from RGP to VGP is also associated with overexpression of the angiogenic factor P10145 . Additionally , the transition of melanoma cells from RGP to VGP is associated with overexpression of the transcription factors CREB and P39905 -1 , both of which may act as survival factors for human melanoma cells . Inactivation of CREB/ P39905 -1 activities in metastatic melanoma cells by dominant-negative CREB or by anti- P39905 -1 single chain antibody fragment ( ScFv ) , resulted in deregulation of P08253 and P43121 / P43121 , increased the sensitivity of melanoma cells to apoptosis , and inhibition of their tumorigenicity and metastatic potential in vivo . In this prospect article , we summarize our data on the role of P05549 and CREB/ P39905 -1 in the progression of human melanoma and report on the development of new fully human antibodies anti- P43121 / P43121 and anti- P10145 which could serve as new modalities for the treatment of melanoma . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . Identification and quantification of dopamine receptor 2 in human eutopic and ectopic endometrium : a novel molecular target for endometriosis therapy . Previous studies in an experimental mouse model of endometriosis have shown that the dopamine agonist ( DA ) cabergoline ( Cb2 ) reduces angiogenesis and endometriotic lesions , hypothetically binding to the dopamine receptor type-2 ( P14416 ) . To date , this has not been described in human endometrium and/or endometriotic lesions . Thus , we aimed to investigate the presence of P14416 in said tissues . Endometrium fragments were implanted in nude mice treated with different doses of Cb2 . Polymerase chain reaction assays and immunohistochemistry were performed to analyze the gene and protein expressions ( respectively ) of P14416 , P15692 , and P15692 receptor-2 ( P35968 ) . In addition , lesions and endometrium from women with mild and severe endometriosis and endometrium from healthy women were collected to analyze their gene expression profile . In experimental endometriosis , P14416 was expressed at gene and protein levels in all three groups . P15692 gene and protein expressions were significantly lower in lesions treated with Cb2 than in controls . P35968 protein expression was significantly lower in experimental lesions treated with Cb2 than in controls . In eutopic endometria , there was a significant decrease in P14416 expression and an increase in P15692 in women with mild and severe endometriosis with respect to healthy patients . In endometriosis , P35968 expression was significantly higher in red than in white and black lesions . P15692 expression was significantly lower in black than in red lesions . P14416 is present in the human eutopic and ectopic endometrium and is regulated by DA , which provides the rationale for pilot studies to explore its use in the treatment of endometriosis . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . 1-(3-Trifluoromethylphenyl) piperazine ( TFMPP ) in the ventral tegmental area reduces the effect of desipramine in the forced swimming test in rats : possible role of serotonin receptors . 1-(3-Trifluoromethylphenyl)piperazine ( TFMPP ) , a serotonin1 ( 5-HT1 ) receptor agonist , injected i.p. in doses of 0.1 and 0.6 mg/kg , did not modify the immobility time of rats in the forced swimming test but significantly antagonized the effect of a 7 days treatment with 10 mg/kg per day desipramine ( DB01151 ) . A similar effect was found on infusing 1 and 5 micrograms/microliters TFMPP bilaterally into the ventral tegmental area ( VTA ) . Infusion of 5 micrograms/microliters TFMPP into the nucleus accumbens or into the globus pallidus did not modify the effect of DB01151 . The effect of 5 micrograms TFMPP infused into the VTA was prevented by the i.p. administration of 5 mg/kg metergoline , a non-selective serotonin receptor antagonist . Infusion of 5 micrograms/microliters 8-hydroxy-2-(di-n-propylamino)tetralin , a specific P08908 receptor agonist , into the VTA did not modify the effect of DB01151 . Besides acting as a P28222 receptor agonist , TFMPP may also act on other 5-HT receptor types , but available evidence suggests that its former action is more important . It thus appears that 5-HT1 receptors in the VTA , presumably of the P28222 type , act by preventing the anti-immobility effect of DB01151 . The role of VTA dopamine and non-dopamine cells in the effect of TFMPP is discussed . Mechanisms of resistance to imatinib and sunitinib in gastrointestinal stromal tumor . Gastrointestinal stromal tumor ( GIST ) , the most common mesenchymal neoplasm of the GI tract and one of the most common sarcomas , is dependent on the expression of the mutated P10721 or platelet-derived growth factor receptor in most cases . Imatinib mesylate potently abrogates the effects of P10721 signaling by directly binding into the DB00171 -binding pocket of the kinase . It is becoming increasingly apparent that the binding affinity of imatinib for the receptor is dependent on the type and location of mutation . Within P10721 , patients whose tumor has an exon 9 mutation are treated by many clinicians with higher doses of imatinib than those patients with mutations within exon 11 . Additionally , there are over 400 unique mutations within exon 11 that may have distinctly different binding affinity for imatinib as well as other kinases . Secondary P10721 mutations generally occur at a codon where imatinib binds resulting in P10721 reactivation and resistance . DB01268 malate , a second-generation P10721 inhibitor is active in imatinib-resistant disease and is FDA-approved for use in this setting . In this review , we describe the biology of the genes and gene mutations responsible for GIST and discuss known and potential clinical implications . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . DB00175 induces rat aortic endothelial cell proliferation and migration via activation of PI3K/Akt/ P42345 / P08133 S6 kinase signaling . The P04035 inhibitors ( statins ) have been shown to exert several vascular protective effects that are not related to changes in cholesterol profile , and these effects of statins are partly caused by the activation of angiogenesis . Endothelial cell ( EC ) proliferation and migration are crucial events for angiogenesis and statins are known to enhance these events . However , the molecular mechanism by which statins promote EC proliferation and migration is not fully understood . In this study , we show Akt and its downstream target mammalian target of rapamycin ( P42345 ) play an important role in pravastatin-induced EC proliferation and migration . We found that pravastatin significantly enhanced the proliferation and migration of rat aortic endothelial cells ( rAECs ) . The addition of pravastatin to rAECs resulted in rapid phosphorylation of Akt and P08133 S6 kinase ( p70S6K ) . LY294002 , a specific inhibitor of phosphatidylinositol 3-kinase ( PI3K ) , blocked both Akt and p70S6K phosphorylation , whereas rapamycin , a specific inhibitor of P42345 , suppressed only p70S6K phosphorylation induced by pravastatin . Furthermore , both LY294002 and rapamycin inhibited pravastatin-induced rAEC proliferation and migration . Taken together , our findings indicate that pravastatin activates PI3K/Akt/ P42345 /p70S6K signaling in this sequential manner and this pathway contributes to pravastatin-induced rAEC proliferation and migration . Wild-type O75581 inhibits , whereas atherosclerosis-linked LRP6R611C increases PDGF-dependent vascular smooth muscle cell proliferation . Vascular smooth muscle cell ( VSMC ) proliferation is an important event in atherosclerosis and other vasculopathies . PDGF signaling is a key mediator of SMC proliferation , but the mechanisms that control its activity remain unclear . We previously identified a mutation in P01130 -related protein 6 ( O75581 ) , O75581 (R611C) , that causes early atherosclerosis . Examination of human atherosclerotic coronary arteries showed markedly increased expression of O75581 and colocalization with PDGF receptor β ( P09619 -β ) . Further investigation showed that wild-type O75581 inhibits but O75581 (R611C) promotes VSMC proliferation in response to PDGF . We found that wild-type O75581 forms a complex with P09619 -β and enhances its lysosomal degradation , functions that are severely impaired in O75581 (R611C) . Further , we observed that wild-type and mutant O75581 regulate cell-cycle activity by triggering differential effects on PDGF-dependent pathways . These findings implicate O75581 as a critical modulator of PDGF-dependent regulation of cell cycle in smooth muscle and indicate that loss of this function contributes to development of early atherosclerosis in humans . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Biomarkers to predict response to sunitinib therapy and prognosis in metastatic renal cell cancer . DB01268 is an orally-administered , multitargeted tyrosine kinase inhibitor . The main targets are vascular endothelial growth factor receptor ( VEGFR ) -1 , P35968 , P35916 , platelet-derived growth factor receptor ( P09619 ) -α , and P09619 -β . Among therapeutic targeting agents , it is the best available in the USA for patients with metastatic renal cell cancer ( RCC ) . Well-constructed clinical trials have led to the worldwide approval of various agents for RCC . However , in clinical practice , it remains difficult to determine the best treatment strategy with these agents . Therefore , the identification of biomarkers to predict response and side-effects and to select optimal dosages is urgently needed . Potential mechanisms of action and resistance need to be understood in order to make accurate predictions . This article briefly reviews candidate biomarkers of sunitinib therapy in terms of clinical variables , genetic factors , and circulating proteins and endothelial cells . Although further validation and implementation is necessary , if validated , biomarkers will help measure the therapeutic response in individual patients and establish treatment strategies for metastatic RCC . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . DB01268 : bridging present and future cancer treatment . Tyrosine kinase receptors ( RTKs ) are a heterogeneous group of transmembrane proteins involved in signal transduction . These receptors are expressed in many different cells and regulate cellular growth , differentiation and angiogenesis . Overexpression and/or the structural alteration of different RTKs classes are generally associated to cancer and , when RTKs-mediated signal transduction pathways are abnormally activated , generate cancer growth , angiogenesis and metastatization . Therapeutic intervention targeting RTKs concerns antagonist drugs as little molecules or monoclonal antibodies . DB01268 malate is a little molecule able to block intracellular tyrosine kinase domain of RTKs , which has both direct anticancer and antiangiogenetic activity . DB01268 targets selectively vascular endothelial growth factor , P10721 , Flt3 and platelet-derived growth factor receptors and the receptor encoded by the ret proto-oncogene . This drug is used in the treatment of gastrointestinal stromal cancer ( GIST ) resistant to imatinib and metastatic renal cell carcinoma ( RCC ) . In this review , we report preclinical data of sunitinib , even about synergism with chemotherapy and radiotherapy , data relative to phase III trials of sunitinib in the treatment of GIST and RCC , and we try to plan what will be future applications of sunitinib in different types of cancer , even in association to chemotherapy , radiotherapy and monoclonal antibodies . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . Experience in the use of sunitinib given as a single agent in metastatic chemoresistant and castration-resistant prostate cancer patients . OBJECTIVE : Vascular endothelial growth factor receptor ( VEGFR ) and platelet-derived growth factor receptor ( P09619 ) correlate with poor prognosis in castration-resistant prostate cancer ( CRPC ) . DB01268 has shown activity in CRPC and at the time of this analysis there was no standard therapy for docetaxel-refractory CRPC . METHODS : We present a case series data collection of 19 patients with a median age of 73 years , CRPC and rising prostate-specific antigen ( PSA ) . Patients received sunitinib 37.5 mg continuous daily dose . One cycle comprised a 4-week period . Patients were evaluated by CT scan every 8 weeks and PSA was monitored every 4 weeks . RESULTS : Median Eastern Cooperative Oncology Group ( ECOG ) performance status score was 2 . Patients received a median of two previous treatment lines for the hormone-refractory setting . Baseline median PSA was 280 ng/ml . Patients received a median of 16 weeks of therapy ( 4 - 48+ ) . One patient achieved a partial response ( 5 % ) and 12 ( 66.7 % ) achieved stable disease for at least 3 months according to RECIST criteria . Median progression-free survival was 4 months . PSA declined > 50 % in 5/19 ( 26.3 % ) and stabilized in 7/19 ( 37 % ) patients . Frequent adverse events were grade 3 asthenia ( 21 % ) , grade 3 diarrhea ( 10 % ) and grade 3 hand-foot syndrome ( 15.7 % ) . CONCLUSIONS : Activity with sunitinib was observed in highly pretreated docetaxel-refractory CRPC with acceptable tolerability . Additional studies should confirm the role of antiangiogenic agents in this setting . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . Strategic combination therapy overcomes tyrosine kinase coactivation in adrenocortical carcinoma . BACKGROUND : Coactivation of tyrosine kinase limits the efficacy of tyrosine kinase inhibitors . We hypothesized that a strategic combination therapy could overcome tyrosine kinase coactivation and compensatory oncogenic signaling in patients with adrenocortical carcinoma ( ACC ) . METHODS : We profiled 88 tyrosine kinases before and after treatment with sunitinib in H295R and SW13 ACC cells . The effects of monotherapy and strategic combination regimens were determined by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium ( ie , MTS ) assay . RESULTS : The minimum inhibitory concentrations ( IC(min) ) of sunitinib quenched its primary targets : P36888 , P35968 , and P07949 . In contrast , P29323 , P08631 , Chk2 , P07947 , CREB , MEK , MSK , p38 , P09769 , and P30530 were hyperactivated . Monotherapy with sunitinib or PD98059 at their IC(min) reduced proliferation by 23 % and 19 % , respectively , in H295R cells and by 25 % and 24 % , respectively , in SW13 cells . DB01268 and PD98059 in combination decreased proliferation by 68 % and 64 % in H295R and in SW13 cells , respectively ( P < .05 versus monotherapy ) . The effects of combination treatment exceeded the sum of the effects observed with each individual agent alone . CONCLUSION : We describe the first preclinical model to develop strategic combination therapy to overcome tyrosine kinase coactivation in ACC . Because many tyrosine kinase inhibitors are readily available , this model can be immediately tested in clinical trials for patients with advanced ACC . DB01268 acts primarily on tumor endothelium rather than tumor cells to inhibit the growth of renal cell carcinoma . DB01268 is a broad-spectrum small-molecule inhibitor of receptor tyrosine kinases ( RTK ) that serves as the present standard of care for first-line therapy of advanced clear cell renal cell carcinoma ( ccRCC ) . A full understanding of the targets and mechanism of action of sunitinib in ccRCC treatment remains incomplete . In this study , we evaluated several tumor cell and endothelial targets of sunitinib and investigated which RTK(s) may specifically contribute to its therapeutic effects . Microarray expression profiling and Western blot analysis revealed that among known sunitinib targets , only platelet-derived growth factor receptor-beta and vascular endothelial growth factor receptor-2 ( P35968 ) were overexpressed in ccRCCs relative to normal tissues . DB01268 was unable to inhibit survival or proliferation of ccRCC cells at pharmacologically relevant concentrations ( approximately 0.1 micromol/L ) that inhibit RTK targets . In contrast , sunitinib inhibited endothelial cell proliferation and motility at the same concentrations by suppressing P35968 signaling . Moreover , whereas sunitinib inhibited the growth of ccRCC xenograft tumors and decreased tumor microvessel density as soon as 12 hours after treatment , sunitinib showed no significant effects on tumor cell proliferation or apoptosis up to 72 hours after treatment . Our findings indicate that sunitinib inhibits ccRCC growth primarily through an antiangiogenic mechanism and not through direct targeting of ccRCC tumor cells . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Pharmacological management of gastrointestinal stromal tumours : an update on the role of sunitinib . The efficacy and tolerability of the receptor tyrosine kinase inhibitor , sunitinib malate , have been demonstrated in phase I-III clinical trials of patients with imatinib-resistant or imatinib-intolerant gastrointestinal stromal tumours ( GIST ) as well as in a worldwide expanded-access study and in a continuous daily dosing ( CDD ) trial . Tumour genotype may have a significant influence on the activity of sunitinib in patients with imatinib-resistant GIST . DB01268 activity was observed across different GIST genotypes and particularly in patients with wild-type and P10721 exon 9 mutations ( all relatively resistant to standard-dose imatinib ) and in patients with secondary P10721 exons 13 and 14 mutations . Adverse events with sunitinib were generally mild to moderate and easily managed by dose reduction , dose interruption or standard supportive measures . Treatment discontinuation can be avoided in most patients by close monitoring before and during treatment with appropriate adverse event management as necessary . The correlation between treatment exposure and clinical response is prompting the search for new approaches to treatment optimisation to ensure that patients derive maximum benefit from sunitinib therapy , including dose adjustments based on blood testing to ensure optimal drug exposure , and the use of the alternative CDD regimen to avoid treatment interruption . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . In vivo antitumor and antimetastatic activity of sunitinib in preclinical neuroblastoma mouse model . Neuroblastoma ( NB ) is one of the most common pediatric solid tumors originating from the neural crest lineage . Despite intensive treatment protocols including megatherapy with hematopoietic stem cell transplantation , the prognosis of NB patients remains poor . More effective therapeutics are required . High vascularity has been described as a feature of aggressive , widely disseminated NB . Our previous work demonstrated the overexpression of vascular endothelial growth factor ( P15692 ) in NB , and we showed that an anti- P15692 receptor ( P35968 ) antibody could induce sustained NB tumor suppression and regression . DB01268 is a kinase inhibitor targeting platelet-derived growth factor receptors and VEGFRs and , therefore , a promising antiangiogenic agent . In this study , we investigated the antitumor activity of sunitinib and its synergistic cytotoxicity with conventional ( cyclophosphamide ) and novel ( rapamycin ) therapies . Both NB cell lines and tumor-initiating cells from patient tumor samples were used in our in vitro and in vivo models for these drug testing . We show that sunitinib inhibits tumor cell proliferation and phosphorylation of VEGFRs . It also inhibits tumor growth , angiogenesis , and metastasis in tumor xenograft models . Low-dose sunitinib ( 20 mg/kg ) demonstrates synergistic cytotoxicity with an P42345 inhibitor , rapamycin , which is more effective than the traditional chemotherapeutic drug , cyclophosphamide . These preclinical studies provide the evidence of antitumor activity of sunitinib both in the early stage of tumor formation and in the progressive metastatic disease . These studies also provide the framework for clinical trial of sunitinib , alone and in combination with conventional and novel therapies to increase efficacy and improve patient outcome in NB . Gene expression profiling in clinically localized prostate cancer : a four-gene expression model predicts clinical behavior . PURPOSE : New diagnostic and prognostic molecular markers are required for prostate cancer , one of the most common male malignancies in Western countries . Gene expression profiling may help to identify genes involved in prostate carcinogenesis , yield clinical biomarkers , and improve tumor classification . EXPERIMENTAL DESIGN : To identify fundamental differences between normal and neoplastic prostate tissue , we used real-time quantitative RT-PCR assays to quantify the mRNA expression of 291 selected genes in samples of normal prostate and of well-documented primary , clinically localized prostate tumors . RESULTS : Forty-six genes showed significantly different expression in tumors relative to normal prostate . The dysregulated genes belong notably to the extracellular membrane and extracellular membrane remodeling categories and are involved in angiogenesis . Furthermore , we obtained a four-gene ( Q9Y5Y7 / Q9Y5Y7 , P01215 , P25116 /PAR1 , and BCL-G ) model that discriminated between the seven patients with and the seven patients without relapse , independently of stage and grade . CONCLUSIONS : Some dysregulated genes are good candidates for use as molecular markers and/or therapeutic targets . Furthermore , differential gene expression profiling of clinically localized prostate tumors from relapsing and nonrelapsing patients identified a set of four genes with a pattern of expression that defines a molecular signature that could predict the clinical behavior of this disease . Inhibition of proliferation and migration of luminal and claudin-low breast cancer cells by P09619 inhibitors . BACKGROUND : Platelet-derived growth factors ( PDGFs ) bind to two receptors , PDGFRα and PDGFRβ to mediate cell proliferation , migration and survival . Although epithelial cells typically do not express high levels of PDGFRs , their expression has been reported to increase in breast cancer cells that have undergone epithelial to mesenchymal transition . METHODS : P09619 signaling was inhibited using DB01268 malate , Imatinib mesylate or DB08896 in murine and human luminal-like and claudin-low mammary tumor cell lines or Masitinib in only the human cell lines . A scratch wound assay was used to assess tumor cell migration while immunofluorescence for phosphorylated histone H3 or cleaved caspase 3 was used to determine tumor cell proliferation and apoptosis , respectively . RESULTS : DB01268 and DB08896 , but not Imatinib , were capable of significantly inhibiting the migration of both murine and human luminal-like and claudin-low breast cancer cells while Masitinib inhibited migration in both human breast cancer cell lines . DB01268 but not DB08896 or Imatinib also significantly suppressed tumor cell proliferation in all four cell lines tested while Masitinib had no significant effect on human breast cancer cell proliferation . None of the P09619 inhibitors consistently regulated mammary tumor cell apoptosis . CONCLUSION : DB01268 , DB08896 and Masitinib may prove clinically useful in inhibiting breast cancer cell migration and metastasis while only DB01268 ( and possibly DB08896 in some breast cancer subtypes ) is effective at inhibiting both migration and proliferation of breast cancer cells . 20-HETE in neovascularization . Cytochrome P450 4A/F ( CYP4A/F ) converts arachidonic acid ( AA ) to 20-HETE by ω-hydroxylation . The contribution of 20-HETE to the regulation of myogenic response , blood pressure , and mitogenic actions has been well summarized . This review focuses on the emerging role of 20-HETE in physiological and pathological vascularization . 20-HETE has been shown to regulate vascular smooth muscle cells ( VSMC ) and endothelial cells ( EC ) by affecting their proliferation , migration , survival , and tube formation . Furthermore , the proliferation , migration , secretion of proangiogenic molecules ( such as HIF-1α , P15692 , SDF-1α ) , and tube formation of endothelial progenitor cells ( EPC ) are stimulated by 20-HETE . These effects are mediated through c-Src- and P00533 -mediated downstream signaling pathways , including MAPK and PI3K/Akt pathways , P29474 uncoupling , and NOX/ROS system activation . Therefore , the CYP4A/F-20-HETE system may be a therapeutic target for the treatment of abnormal angiogenic diseases . Mutation D816V alters the internal structure and dynamics of c- P10721 receptor cytoplasmic region : implications for dimerization and activation mechanisms . The type III receptor tyrosine kinase ( RTK ) P10721 plays a crucial role in the transmission of cellular signals through phosphorylation events that are associated with a switching of the protein conformation between inactive and active states . D816V P10721 mutation is associated with various pathologies including mastocytosis and cancers . D816V-mutated P10721 is constitutively active , and resistant to treatment with the anti-cancer drug Imatinib . To elucidate the activating molecular mechanism of this mutation , we applied a multi-approach procedure combining molecular dynamics ( MD ) simulations , normal modes analysis ( Q13145 ) and binding site prediction . Multiple 50-ns MD simulations of wild-type P10721 and its mutant D816V were recorded using the inactive auto-inhibited structure of the protein , characteristic of type III RTKs . Computed free energy differences enabled us to quantify the impact of D816V on protein stability in the inactive state . We evidenced a local structural alteration of the activation loop ( A-loop ) upon mutation , and a long-range structural re-organization of the juxta-membrane region ( JMR ) followed by a weakening of the interaction network with the kinase domain . A thorough normal mode analysis of several MD conformations led to a plausible molecular rationale to propose that JMR is able to depart its auto-inhibitory position more easily in the mutant than in wild-type P10721 and is thus able to promote kinase mutant dimerization without the need for extra-cellular ligand binding . Pocket detection at the surface of Q13145 -displaced conformations finally revealed that detachment of JMR from the kinase domain in the mutant was sufficient to open an access to the catalytic and substrate binding sites . New perspectives : role of DB01268 in breast cancer . DB01268 malate ( SU11248 ) is a multitarget oral tyrosine kinase receptor ( RTKs ) inhibitor which was approved by FDA in renal cells carcinoma ( RCC ) and imatinib-resistant or imatinib-intollerant gastro-intestinal stromal tumour ( GIST ) . DB01268 is able to inhibit RTKs such as receptors for platelet-derived growth factor ( PDGF-Rα and β ) and for vascular endothelial growth factor ( VEGFRs ) . It is able to inhibit P10721 receptor , colony stimulating factor type 1 receptor ( P07333 ) , glial cell line neutrophic factor receptor ( P07949 ) , fms-like tyrosine kinase receptor-3 ( P36888 or P36888 ) , signal transducer and activator of transcription 3 ( P40763 ) and AKT ( protein kinase B ) in tumour cells . Many DB01268 targets play important roles in growth and survival of human breast cancer ( BC ) . The " rationale " of DB01268 in BC ( with or without others antiagiogenetic therapy ) is its ability to block simultaneously intracellular portion of RTKs inhibiting many downstream signals . We overviewed the most relevant studies concerning DB01268 in metastatic BC . Crystal structure of the PDZ1 domain of human Na(+)/H(+) exchanger regulatory factor provides insights into the mechanism of carboxyl-terminal leucine recognition by class I PDZ domains . The Na(+)/H(+) exchanger regulatory factor ( O14745 ; also known as O14745 ) contains two PDZ domains that mediate the assembly of transmembrane and cytosolic proteins into functional signal transduction complexes . The O14745 PDZ1 domain interacts specifically with the motifs DSLL , DSFL , and DTRL present at the carboxyl termini of the beta(2) adrenergic receptor ( beta(2)AR ) , the platelet-derived growth factor receptor ( P09619 ) , and the cystic fibrosis transmembrane conductance regulator ( P13569 ) , respectively , and plays a central role in the physiological regulation of these proteins . The crystal structure of the human O14745 PDZ1 has been determined at 1.5 A resolution using multiwavelength anomalous diffraction phasing . The overall structure is similar to known PDZ structures , with notable differences in the O14745 PDZ1 carboxylate-binding loop that contains the GYGF motif , and the variable loop between the beta2 and beta3 strands . In the crystalline state , the carboxyl-terminal sequence DEQL of PDZ1 occupies the peptide-binding pocket of a neighboring PDZ1 molecule related by 2-fold crystallographic symmetry . This structure reveals the molecular mechanism of carboxyl-terminal leucine recognition by class I PDZ domains , and provides insights into the specificity of O14745 interaction with the carboxyl termini of several membrane receptors and ion channels , including the beta(2)AR , P09619 , and P13569 . Circulating cytokines and monocyte subpopulations as biomarkers of outcome and biological activity in sunitinib-treated patients with advanced neuroendocrine tumours . BACKGROUND : DB01268 is approved worldwide for treatment of advanced pancreatic neuroendocrine tumours ( pNET ) , but no validated markers exist to predict response . This analysis explored biomarkers associated with sunitinib activity and clinical benefit in patients with pNET and carcinoid tumours in a phase II study . METHODS : Plasma was assessed for vascular endothelial growth factor ( P15692 ) -A , soluble P15692 receptor ( sVEGFR ) -2 , sVEGFR-3 , interleukin ( IL ) -8 ( n=105 ) , and stromal cell-derived factor ( SDF ) -1α ( n=28 ) . Pre-treatment levels were compared between tumour types and correlated with response , progression-free ( PFS ) , and overall survival ( OS ) . Changes in circulating myelomonocytic and endothelial cells were also analysed . RESULTS : Stromal cell-derived factor-1α and sVEGFR-2 levels were higher in pNET than in carcinoid ( P=0.003 and 0.041 , respectively ) . High ( above-median ) baseline SDF-1α was associated with worse PFS , OS , and response in pNET , and high sVEGFR-2 with longer OS ( P⩽0.05 ) . For carcinoid , high P10145 , sVEGFR-3 , and SDF-1α were associated with shorter PFS and OS , and high P10145 and SDF-1α with worse response ( P⩽0.05 ) . Among circulating cell types , monocytes showed the largest on-treatment decrease , particularly P08571 + monocytes co-expressing P17948 or P61073 . CONCLUSIONS : P10145 , sVEGFR-3 , and SDF-1α were identified as predictors of sunitinib clinical outcome . Putative pro-tumorigenic P61073 + and P17948 + monocytes represent novel candidate markers and biologically relevant targets explaining the activity of sunitinib . Targeting lymphangiogenesis after islet transplantation prolongs islet allograft survival . BACKGROUND : Lymphatics are important for their conduit functions of transporting antigen , immune cells , and inflammatory mediators to draining lymph nodes and to the general circulation . Lymphangiogenesis is involved in many pathologic processes ; however , the roles for lymphatic responses in transplantation have not been thoroughly investigated . METHODS : Mice were made diabetic by a single high dose of streptozotocin and then received islet allografts . Animals were treated with three different lymphatic inhibitors . FTY720 , an analog of sphingosine 1-phosphate , inhibited lymphocyte migration into afferent and efferent lymphatics . DB01268 , a kinase inhibitor , blocked several receptors , including vascular endothelial growth factor receptor 3 ( P35916 ) , the major growth factor receptor for lymphatic endothelial cells . Anti- P35916 monoclonal antibody specifically inhibited P35916 . Diabetes was determined by daily monitoring of blood glucose levels . Inflammation within islet grafts was assessed by immunohistochemistry for insulin , T cells ( CD3 ) , and lymphatics ( Q9Y5Y7 ) . RESULTS : After transplantation , lymphangiogenesis occurred in islet allografts and in draining lymph nodes . FTY720 , sunitinib , and anti- P35916 each inhibited lymphangiogenesis in the islets and significantly prolonged allograft survival . Immunofluorescent staining demonstrated that administration of each of the lymphatic inhibitors resulted in preservation of islets and β-cells along with a markedly reduced infiltration of T cells into the grafts . CONCLUSION : Lymphangiogenesis occurs in islet allografts in response to inflammation and plays a key role in the islet inflammation in alloimmunity . Interfering with lymphatic function leads to inhibition of lymphangiogenesis and prolonged or indefinite allograft survival . These observations suggest new therapeutic targets for rejection and tolerance . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . The role of Smad signaling in vascular and hematopoietic development revealed by studies using genetic mouse models . Smads are intracellular mediators of transforming growth factor beta ( TGF-beta ) superfamily signaling . In this review , we focus on the genetic mouse models for Smad pathways , which have provided functional evidence regarding the complex circuitry in angiogenesis and hematopoiesis during development . In the early stages of vascular development , TGF-beta signaling is a contributing factor in angiogenesis and vascular maturation . Whereas in the later embryogenesis , selected molecules of Smad pathways , such as P37173 ( TbRII ) , P36897 , and Q99717 , seem to be dispensable for vessel morphogenesis and integrity . TGF-beta signaling is not required in the induction of hematopoietic precursors from mesoderm , but inhibits the subsequent expansion of committed hematopoietic precursors . By contrast , bone morphogenetic protein 4 ( P12644 ) has long been acknowledged pivotal in mesoderm induction and hematopoietic commitment during development . However , recent genetic evidence shows the P12644 - P36894 axis is not crucial for the formation of hematopoietic cells from P35968 (+) mesoderm . Because of the highly redundant mechanisms within the Smad pathways , the precise role of the Smad signaling involved in vascular and hematopoietic development remains nebulous . The generation of novel cell lineage restricted Cre transgenes would shed new light on the future relevant investigations . [ Evidence-based treatment of gastrointestinal stromal tumor ( GIST ) with tyrosine kinase inhibitors-imatinib and sunitinib ] . Gastrointestinal stromal tumors ( GIST ) are sarcoma in the gastrointestinal tract which may be caused by somatic gain-of-function mutations in the P10721 or P16234 gene . Imatinib mesylate has shown significant safety and great effectiveness for patients with P10721 -positive unresectable , advanced , or metastatic GIST . The response rate ( RR ) was no less than 50 % , disease control rate ( DCR ) of more than 85 % , and the median progression-free survival ( PFS ) has been nearly two years . Also , imatinib has been shown to improve the prognosis of GIST patients . Meanwhile , sunitinib malate , used for patients with imatinib-resistant GIST or imatinib-intolerant patients , has also shown modest tolerability and fairly good outcomes . DB01268 shows a less than 10 % RR , a 30 % to 40 % DCR , and a median of nearly 8 months of PFS . Molecularly targeted therapy has prolonged overall survival of advanced GIST patients from 1 . 5 years in the pre-imatinib era to 5 years after the introduction of imatinib . There remains , however , a great unmet medical need for new agents and/or multidisciplinary treatments for advanced GIST patients . A decade of tyrosine kinase inhibitor therapy : Historical and current perspectives on targeted therapy for GIST . The introduction of molecularly targeted therapies has ushered in a considerable transformation in the management of gastrointestinal stromal tumors ( GIST ) that currently defines the paradigm of targeted therapy for solid tumors . Indeed , in the past decade the management of GIST has evolved from a disease only effectively treatable by surgery to the archetype of a tumor treatable with a molecularly targeted therapy . Better understanding of the molecular and genetic characteristics that underlie the aberrant behavior of GIST has increased the accuracy of its diagnosis and allowed for the identification of distinct genetic hallmarks , prognostic groups , and treatment strategies . Collectively , this has resulted in the development of the targeted tyrosine kinase inhibitors ( TKIs ) imatinib and sunitinib , and continues to prompt studies of novel agents in this disease . Since approval in 2002 , imatinib has been shown to provide a high level of clinical efficacy in patients with advanced GIST , including a median progression-free survival ( PFS ) of 2 years and median overall survival approaching 5 years , with some patients progression-free after 10 years of treatment . Imatinib is now also approved in adult patients following resection of P10721 -positive GIST . In 2006 , sunitinib was approved for the treatment of advanced GIST after failure of imatinib . DB01268 provides significant benefit in this setting , with a median PFS close to 6 months after imatinib failure . Following progression on these agents , patients have limited treatment options . This critical unmet need is being addressed by the development of new TKIs and the use of novel regimens with approved agents . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Progenitor cells harvested from bovine follicles become endothelial cells . Hematopoietic-like colonies develop in post-confluent granulosa cell cultures derived from bovine antral follicles . Previously , we had shown that these colonies gave rise to macrophages . In the present study , we validated the presence of somatic P10721 -positive ( P10721 (+) ) progenitor cells in colony-containing granulosa cell cultures . The cultures expressed the progenitor cell markers Sox-2 , Oct 3/4 , P10721 , and alkaline phosphatase in western blot analysis . The successful double immunofluorescence localization of P10721 and P08571 , P08575 , CD133 , or P15692 -R2 revealed a specific subpopulation of progenitor cells . Flow cytometry showed that cells doubly positive for P10721 and P08571 or P08575 comprised less than 10 % of the population . The P10721 (+) cells were purified by magnetic selection and differentiated with the hanging drop technique using haematopoietic differentiation medium . Pure cultures of either granulosa cells or endothelial cells were obtained . The spindle-shaped and epithelioid phenotypes indicated endothelial cell heterogeneity of microvascular source . We conclude that progenitor cells are obtained from the follicle harvest , which differentiate into endothelial cells . The cells are relevant for findings to angiogenesis and luteinization of the corpus luteum .

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interacts_with DB00831?

multiple_choice

Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . [ P62158 -dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium ] . Highly purified plasma membrane ( PM ) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein . Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3 % of the original level . The activity of Ca , Mg-ATPase is thereby decreased by 40 % . Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour . The maximal activation of Ca , Mg-ATPase is observed with 10(-7) M calmodulin . P62158 increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity . DB00831 ( 20 microM ) diminishes the activating effect of exogenous calmodulin on Ca , Mg-ATPase . P62158 stimulates Ca , Mg-ATPase at low concentrations of Ca2+ ( 10(-8)-10(-6) M ) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax -- from 0,8 to 1.42 mumol Pl/mg protein/hour . It is supposed that the activating effect of calmodulin on Ca , Mg-ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme . P62158 interacts with the platelet ADP receptor P47900 . P47900 [ P2 ( purinergic type-2 ) -receptor 1 ] is a G-protein-coupled ADP receptor that regulates platelet activation and ADP-induced Ca2+ signalling . Studies using P47900 -knockout mice , G(q)-deficient mice or P47900 -selective inhibitors have previously identified a key role for P47900 in pathophysiological thrombus formation at high shear stress . We provide evidence that a positively charged juxtamembrane sequence within the cytoplasmic C-terminal tail of P47900 can bind directly to the cytosolic regulatory protein calmodulin . Deletion by mutagenesis of the calmodulin-binding domain of P47900 inhibits intracellular Ca2+ flux in transfected cells . These results suggest that the interaction of calmodulin with the P47900 C-terminal tail may regulate P47900 -dependent platelet aggregation . Skinned coronary smooth muscle : calmodulin , calcium antagonists , and DB02527 influence contractility . The effects of Ca2+ , calmodulin , DB02527 , the catalytic subunit of DB02527 -dependent protein kinase ( CSU ) and some Ca2+ antagonists were studied in chemically ( Triton X-100 ) skinned coronary smooth muscle . P62158 increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration . DB00831 , a calmodulin antagonist , inhibited Ca2+-calmodulin-induced contraction . Both DB02527 and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration . DB08980 , a Ca2+-antagonist , at 2 x 10(-4) M produced a significant inhibitory effect , which was reduced by increasing the Ca2+ concentration . From other Ca2+ antagonists tested , W-7 , but not D600 and verapamil , produced some inhibitory effect . The data indicate that the response of skinned coronary smooth muscle to Ca2+ , calmodulin and DB02527 are similar to those obtained with other skinned smooth muscles . Furthermore , skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle . Gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides by cDNA microarrays . To investigate the gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides ( ODNs ) , Q15761 antisense , mismatched ODNs or vehicle was intracerebroventricularly injected and cDNA microarrays were undertaken . Central administration of Q15761 antisense ODNs decreased food intake , body weight and serum insulin compared with both vehicle and mismatched ODNs . The average area of adipocytes both at retroperitoneal and epididymal adipose tissue were fall in antisense group while only the weight of the retroperitoneal fat pats was reduced in antisense group . cDNA microarrays containing 18,000 genes/Ests were used to investigate gene expression of adipose tissue . Autoradiographic analysis showed that 404 , 81 , and 34 genes were differently expressed over twofold , threefold , and fivefold , respectively . The analysis of gene expression profiles indicated that 332 genes were up-regulated and 187 genes were down-regulated in response to Q15761 antisense ODNs treatment . Different clusters of genes associated with apoptosis , signal transduction , energy metabolism , lipid metabolism , etc. , such as P51114 , Q8WV24 , Q7L5Y9 , P27986 , P13598 , Q00169 , P62158 , Q13557 , P61925 , P14416 , O95258 , CKB , P22760 , P38571 , O15254 , O60427 , were concerned . Analysis of differentially expressed genes will help to understand the effects of Q15761 antisense ODNs therapy . Establishment and phenotypic characterization of human U937 cells with inducible P210 P11274 / P00519 expression reveals upregulation of P13688 ( CD66a ) . Chronic myeloid leukemia ( CML ) is characterized by the expression of the P210 P11274 / P00519 fusion protein . The molecular mechanisms behind this oncogene-mediated hematological disease are , however , not fully understood . Here , we describe the establishment and phenotypic characterization of U937 cells in which P210 P11274 / P00519 can be conditionally expressed using tetracycline . The induction of P11274 / P00519 in the obtained clones resulted in a rapid phosphorylation of the P42224 , P40763 and P42229 molecules , consistent with the findings in other model systems . Phenotypic characterization of the clones revealed that P11274 / P00519 induces a slight decrease in the proliferation and viability , without a marked effect on cell cycle distribution , the rate of apoptosis or on cellular differentiation , as judged by several cell surface markers and capacity to reduce nitro blue tetrazolium . Interestingly , P11274 / P00519 was found to upregulate the expression of carcinoembryonic-related antigen ( P06731 ) P62158 ( CD66a ) , which is a plasma membrane-linked glycoprotein belonging to the CEAs and involved in signal transduction and cellular adhesion . The expression of P13688 was reversible upon imatinib treatment in P11274 / P00519 -expressing U937 cells as well as in P11274 / P00519 -positive K562 cells . The established cell lines may prove useful in further modeling and dissection of P11274 / P00519 -induced leukemogenesis . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . Mutation of the calmodulin binding motif IQ of the L-type Ca(v)1.2 Ca2+ channel to EQ induces dilated cardiomyopathy and death . Cardiac excitation-contraction coupling ( EC coupling ) links the electrical excitation of the cell membrane to the mechanical contractile machinery of the heart . DB01373 channels are major players of EC coupling and are regulated by voltage and Ca(2+)/calmodulin ( P62158 ) . P62158 binds to the IQ motif located in the C terminus of the Ca(v)1.2 channel and induces Ca(2+)-dependent inactivation ( CDI ) and facilitation ( P05231 ) . Mutation of DB00167 to DB00142 ( Ile1624Glu ) in the IQ motif abolished regulation of the channel by CDI and P05231 . Here , we addressed the physiological consequences of such a mutation in the heart . Murine hearts expressing the Ca(v)1.2(I1624E) mutation were generated in adult heterozygous mice through inactivation of the floxed WT Ca(v)1.2( Q401N2 ) allele by tamoxifen-induced cardiac-specific activation of the MerCreMer Cre recombinase . Within 10 days after the first tamoxifen injection these mice developed dilated cardiomyopathy ( DCM ) accompanied by apoptosis of cardiac myocytes ( CM ) and fibrosis . In Ca(v)1.2(I1624E) hearts , the activity of phospho- P62158 kinase II and phospho-MAPK was increased . CMs expressed reduced levels of Ca(v)1.2(I1624E) channel protein and I(Ca) . The Ca(v)1.2(I1624E) channel showed " CDI " kinetics . Despite a lower sarcoplasmic reticulum Ca(2+) content , cellular contractility and global Ca(2+) transients remained unchanged because the EC coupling gain was up-regulated by an increased neuroendocrine activity . Treatment of mice with metoprolol and captopril reduced DCM in Ca(v)1.2(I1624E) hearts at day 10 . We conclude that mutation of the IQ motif to IE leads to dilated cardiomyopathy and death . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Anti-angiogenic effects and mechanisms of polysaccharides from Antrodia cinnamomea with different molecular weights . ETHNOPHARMACOLOGICAL RELEVANCE : Antrodia cinnamomea is a popular medicinal mushroom in Taiwan that has been widely used for treatment of various cancers and liver diseases . AIM OF THE STUDY : This study aimed to investigate the immunomodulatory effect on angiogenesis of polysaccharides from mycelia of Antrodia cinnamomea ( PMAC ) . MATERIALS AND METHODS : PMAC were extracted in boiling water , precipitated with 95 % ethanol , and separated into four different molecular weights ( < 5 , 5-30 , 30-100 , > 100 kDa ) . Tube formation and chorioallantoic membrane ( P62158 ) assay were used to determine the in vitro and ex vivo anti-angiogenic effects . RESULTS : Only the PMAC-mononuclear cells ( MNCs ) -conditioned medium ( CM ) with MW > 100 kDa significantly and concentration-dependently decreased the secretion of vascular endothelial growth factor in human leukemia cells and inhibited the matrigel tube formation in human umbilical vein endothelial cells . Similarly only the PMAC-MNC-CM with MW > 100 kDa significantly and concentration-dependently increased the levels of interleukin ( IL ) -12 and interferon-gamma ( P01579 ) . In addition , the ex vivo P62158 assay revealed that only the PMAC with MW > 100 kDa significantly and dose-dependently inhibited neovascularization . CONCLUSIONS : PMAC with MW > 100 kDa are anti-angiogenic in vitro and ex vivo , and the effects are likely through immunomodulation . Altered transcriptional regulators in response to serum in immortalized lymphocytes from Alzheimer 's disease patients . Cell cycle disturbances may precede neuronal death in Alzheimer 's disease ( AD ) . We described alterations , in lymphocytes from AD patients , on the activity of two transcription factors , E2F and NF-kappaB , involved in cell proliferation and survival regulation , demonstrating that cell cycle dysfunction also occurs in peripheral cells . The analysis of E2F-DNA binding activity revealed lower signal intensity of protein-DNA complexes in AD cells , which correlated with increased phosphorylation of retinoblastoma ( P06400 ) related proteins and enhanced proliferation . The calmodulin ( P62158 ) antagonist calmidazolium ( DB01489 ) abrogated the increased activity of AD cells by partially dephosphorylating P06400 and Q08999 . The NF-kappaB-DNA binding activity increased as cell progress through the cell cycle . The reduced NF-kappaB activation observed in AD cells appears not to be related to the increased phosphorylation of the P06400 family proteins nor with the enhanced proliferative activity of AD cells , but seems to protect them from death induced by the loss of trophic support . Ca2+/ P62158 antagonists rescue NF-kappaB-DNA binding activity and sensitize AD cells to serum withdrawal . These observations suggest that disruption of Ca2+/ P62158 signaling pathway could be linked mechanistically to its pro cell survival actions , promoting enhanced proliferation or decreased cell death depending on the presence of growth-stimulatory signals . Fluorescence energy transfer analysis of calmodulin-peptide complexes . The interactions between calmodulin and the tryptophan residues of synthetic peptides corresponding to the calmodulin binding domains of skeletal muscle myosin light-chain kinase and the plasma membrane calcium pump were examined . The single tryptophan residue contained in each peptide became relatively immobilized and inaccessible to iodide ion upon binding to calmodulin , indicating that the indole side chain was inserted into a hydrophobic cleft in the surface of calmodulin . Fluorescence energy transfer from peptidyl tryptophan residues to an AEDANS moiety attached to cysteine-26 of spinach calmodulin was measured . Included in these analyses was a tryptophan-containing peptide analog of the calmodulin binding domain of neuromodulin . These data indicated that the indole ring of each peptide inserted 32-35 A away from cysteine-26 and may therefore interact with the carboxyl-terminal lobe of P62158 in its " bent " conformation [ Persechini & Kretsinger ( 1988a ) J. Cardiovasc. Pharmacol. 12 ( Suppl 5 ) , S1- P28222 ; Ikura et al. ( 1992 ) Science 256 , 632-638 ; Vorherr et al. ( 1992 ) Eur. J. Biochem. 204 , 931-937 ] . The interchange of tryptophan-3 and phenylalanine-21 of the calcium pump peptide increased the efficiency of energy transfer to the AEDANS-moiety approximately 12-fold , reducing the calculated distance to 20 A . These data suggest that phenylalanine-21 of the calcium pump peptide interacts with the hydrophobic cleft in the amino-terminal lobe of P62158 . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent .

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Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Dynamic and ligand-selective interactions of vitamin D receptor with retinoid X receptor and cofactors in living cells . The vitamin D receptor ( P11473 ) mediates vitamin D signaling in numerous physiological and pharmacological processes , including bone and calcium metabolism , cellular growth and differentiation , immunity , and cardiovascular function . Although transcriptional regulation by P11473 has been investigated intensively , an understanding of ligand-selective dynamic P11473 conformations remains elusive . Here , we examined ligand-dependent dynamic interactions of P11473 with retinoid X receptor ( RXR ) , steroid receptor coactivator 1 ( Q15788 ) , and silencing mediator of retinoic acid and thyroid hormone receptor ( Q9Y618 ) in cells using fluorescence resonance energy transfer ( FRET ) and chromatin immunoprecipitation ( ChIP ) assays . We compared the effects of 1α,25-dihydroxyvitamin D(3) [ 1,25(OH)(2)D(3) ] , lithocholic acid ( LCA ) , and ( 25R ) -25-adamantyl-1α,25-dihydroxy-2-methylene-22,23-didehydro-19,26,27-trinor-20-epivitamin D(3) ( ADTT ) , a partial agonist/antagonist vitamin D derivative . In the absence of ligand , P11473 homodimers were preferred to RXR heterodimers and were associated with Q9Y618 . 1,25(OH)(2)D(3) induced heterodimerization with RXR , dissociation of Q9Y618 , and association of Q15788 . LCA and ADTT induced those effects to a lesser extent at concentrations that did not induce expression of the P11473 target gene Q07973 in human embryonic kidney ( P29320 ) 293 cells . Unlike in HEK293 cells , ADTT increased Q07973 expression in HCT116 cells and increased the association of P11473 and Q9Y618 on the Q07973 promoter . The results indicate that ligand-selective conformation may lead to unique cofactor complex formation in a cell context-dependent manner . The combination of FRET and ChIP assays is a powerful tool useful in understanding ligand-selective dynamic P11473 conformations and the development of selective P11473 modulators . Synthesis of new thiazolo[4,5-d]pyrimidines as DB01285 releasing factor modulators . P06850 ( CRF ) is a neurohormone that plays a crucial role in integrating the body 's overall response to stress . It appears necessary and sufficient for the organism to mount functional , physiological and endocrine responses to stressors . CRF is released in response to various triggers such as chronic stress . The role of CRF and its involvement in these neurological disorders suggest that new drugs that can target the CRF function or bind to its receptors may represent a new development of neuropsychiatric medicines to treat various stress-related disorders including depression , anxiety and addictive disorders . Based on pharmacophore of the CRF1 receptor antagonists , a new series of thiazolo[4,5-d] pyrimidines were synthesized as P06850 ( CRF ) receptor modulators and the prepared compounds carry groups shown to produce optimum binding affinity to CRF receptors . Twenty two compounds were evaluated for their CRF1 receptor binding affinity in P29320 293 cell lines and two compounds 5o and 5s showed approximately 25 % binding affinity to CRF1 receptors . Selected compounds ( 5c and 5f ) were also evaluated for their effect on expression of genes associated with depression and anxiety disorders such as CRF1 , P16220 , P21397 , P31645 , P01303 , DatSLC6a3 , and P09172 and significant upregulation of CRF1 mRNA has been observed with compound 5c . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . The testis anion transporter TAT1 ( Q96RN1 ) physically and functionally interacts with the cystic fibrosis transmembrane conductance regulator channel : a potential role during sperm capacitation . The Slc26 gene family encodes several conserved anion transporters implicated in human genetic disorders , including Pendred syndrome , diastrophic dysplasia and congenital chloride diarrhea . We previously characterized the TAT1 ( testis anion transporter 1 ; Q96RN1 ) protein specifically expressed in male germ cells and mature sperm and showed that in the mouse , deletion of Tat1 caused male sterility due to a lack of sperm motility , impaired sperm capacitation and structural defects of the flagella . Ca(2+) , Cl(-) and HCO(3)(-) influxes trigger sperm capacitation events required for oocyte fertilization ; these events include the intracellular rise of cyclic adenosine monophosphate ( DB02527 ) and protein kinase A ( PKA ) -dependent protein phosphorylation . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is expressed in mature sperm and has been shown to contribute to Cl(-) and HCO(3)(-) movements during capacitation . Furthermore , several members of the SLC26 family have been described to form complexes with P13569 , resulting in the reciprocal regulation of their activities . We show here that TAT1 and P13569 physically interact and that in Xenopus laevis oocytes and in CHO- P04264 cells , TAT1 expression strongly stimulates P13569 activity . Consistent with this , we show that Tat1 inactivation in mouse sperm results in deregulation of the intracellular DB02527 content , preventing the activation of PKA-dependent downstream phosphorylation cascades essential for sperm activation . These various results suggest that TAT1 and P13569 may form a molecular complex involved in the regulation of Cl(-) and HCO(3)(-) fluxes during sperm capacitation . In humans , mutations in P13569 and/or TAT1 may therefore be causes of asthenozoospermia and low fertilizing capacity of sperm . P49841 -mediated tau phosphorylation in cultured cell lines . To study further the role of glycogen synthase kinase-3beta on tau phosphorylation , glycogen synthase kinase-3beta and tau expression vectors were co-transfected into CHO- P04264 , COS-7 and SH-SY5Y cell . Tau phosphorylation was assessed by phosphorylation-dependent antibodies AT-8 , AT-180 , AT-270 and PHF-1 . The AT-270 and AT-8 epitopes were consistently phosphorylated by glycogen synthase kinase-3beta in the three cell lines . Phosphorylation on AT-180 epitope was significant in CHO- P04264 and SH-SY5Y cells while PHF-1 epitope was hyper-phosphorylated only in SH-SY5Y cells . We also found that lithium induces phosphorylation of the serine 9 residue of glycogen synthase kinase-3beta together with inhibition of tau phosphorylation on PHF-1 epitope in all the three cell lines . This suggests a novel mechanism whereby lithium-mediated inhibition of GSK-3beta activity influences tau phosphorylation . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . P60568 inhibits DB01221 receptor-mediated currents directly and may differentially affect subtypes . Using whole-cell patch-clamp recordings , this study investigated the effects of interleukin-2 ( P60568 ) on N-methyl-d-aspartate ( DB01221 ) receptor-mediated currents ( I( DB01221 ) ) in rat cultured hippocampal neurons and human embryonic kidney ( P29320 ) 293 cells expressing recombinant DB01221 receptors . We found that P60568 ( 0.01-1ng/ml ) immediately and significantly decreased peak I( DB01221 ) in cultured neurons . Interestingly , the peak I( DB01221 ) induced in P29320 293 cells was also inhibited by P60568 . We also found that P60568 differentially decreased the peak amplitudes of Q12879 - and Q13224 -containing DB01221 receptor-mediated currents ( I( Q12879 ) and I( Q13224 ) ) by 54+/-5 % and 30+/-4 % , respectively . These results provide new evidence that P60568 induces rapid inhibition of peak currents of DB01221 receptor-mediated responses with possible Q9UHB4 / Q12879 and Q9UHB4 / Q13224 subtype-differentiation , and suggest that the inhibition is mediated by direct interaction between P60568 and DB01221 receptors . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . Potent and selective activation of the pancreatic beta-cell type K( DB00171 ) channel by two novel diazoxide analogues . AIMS/HYPOTHESIS : We investigated the pharmacological properties of two novel DB00171 sensitive potassium ( K( DB00171 ) ) channel openers , 6-Chloro-3-isopropylamino-4 H-thieno [ 3,2- e ] -1,2,4-thiadiazine 1,1-dioxide ( NNC 55-0118 ) and 6-chloro-3-(1-methylcyclopropyl)amino-4 H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide ( NN414 ) , on the cloned cardiac ( Kir6.2/SUR2A ) , smooth muscle ( Kir6.2/SUR2B ) and pancreatic beta cell ( Kir6.2/ Q09428 ) types of K( DB00171 ) channel . METHODS : We studied the effects of these compounds on whole-cell currents through cloned K( DB00171 ) channels expressed in Xenopus oocytes or mammalian cells ( HEK293 ) . We also used inside-out macropatches excised from Xenopus oocytes . RESULTS : In P29320 293 cells , NNC 55-0118 and NN414 activated Kir6.2/ Q09428 currents with EC(50) values of 0.33 micromol/l and 0.45 micromol/l , respectively , compared with that of 31 micro mol/l for diazoxide . Neither compound activated Kir6.2/SUR2A or Kir6.2/SUR2B channels expressed in oocytes , nor did they activate Kir6.2 expressed in the absence of Q09428 . Current activation was dependent on the presence of intracellular MgATP , but was not supported by MgADP . CONCLUSION/INTERPRETATION : Both NNC 55-0118 and NN414 selectively stimulate the pancreatic beta-cell type of K( DB00171 ) channel with a higher potency than diazoxide , by interaction with the Q09428 subunit . The high selectivity and efficacy of the compounds could prove useful for treatment of disease states where inhibition of insulin secretion is beneficial . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . P60568 abolishes myeloid cell accumulation induced by Lewis lung carcinoma . Immune aberration in cancer patients can be at least partly ascribed to an accumulation of immature myeloid cells and monocytes/macrophages with immunosuppressive functions . Mice implanted with Lewis lung carcinoma 2 ( LL/2 ) cells show marked splenomegaly as the tumors progress , and this condition is accompanied by impaired T cell activities . We characterized the cells that accumulated in the spleens of LL/2 tumor-bearing mice and attempted to restore the normal cell population by employing interleukin-2 ( P60568 ) . Flow cytometric analysis revealed that the cells expressing Mac1 , P33681 , NK- P04264 , Gra-1 , and MHC class II antigens on their surfaces drastically decreased in number when LL/2 had been engineered to produce P60568 . P60568 also restored the concanavalin A ( ConA ) -mediated proliferative response and P60568 production of the spleen cells . The in vivo growth of P60568 -producing tumors was significantly slower than that of parental LL/2 cells . Therefore , local P60568 production may reverse systemic immune abnormality by stopping myeloid cell accumulation . Mechanisms of atrial-selective block of Na⁺ channels by ranolazine : I . Experimental analysis of the use-dependent block . Atrial-selective inhibition of cardiac Na(+) channel current ( I(Na) ) and I(Na)-dependent parameters has been shown to contribute to the safe and effective management of atrial fibrillation . The present study examined the basis for the atrial-selective actions of ranolazine . Whole cell I(Na) was recorded at 15°C in canine atrial and ventricular myocytes and in human embryonic kidney ( P29320 ) -293 cells expressing Q14524 . Tonic block was negligible at holding potentials from -140 to -100 mV , suggesting minimal drug interactions with the closed state . Trains of 40 pulses were elicited over a range of holding potentials to determine use-dependent block . Guarded receptor formalism was used to analyze the development of block during pulse trains . Use-dependent block by ranolazine increased at more depolarized holding potentials , consistent with an interaction of the drug with either preopen or inactivated states , but was unaffected by longer pulse durations between 5 and 200 ms , suggesting a weak interaction with the inactivated state . Block was significantly increased at shorter diastolic intervals between 20 and 200 ms . Responses in atrial and ventricular myocytes and in P29320 -293 cells displayed a similar pattern . DB00243 is an open state blocker that unbinds from closed Na(+) channels unusually fast but is trapped in the inactivated state . Kinetic rates of ranolazine interactions with different states of atrial and ventricular Na(+) channels were similar . Our data suggest that the atrial selectivity of ranolazine is due to a more negative steady-state inactivation curve , less negative resting membrane potential , and shorter diastolic intervals in atrial cells compared with ventricular cells at rapid rates . Novel biologically active bibenzyls from Bauhinia saccocalyx Pierre . Four new bibenzyls , bauhinols A-D ( 1-4 ) , together with the two known bibenzyls 5 and 6 , were isolated from the roots of Bauhinia saccocalyx , and their structures were elucidated by analyses of spectroscopic data . Bauhinol A ( 1 ) exhibits significant cytotoxicity towards NCI-H187 ( small-cell lung cancer ) , BC ( breast cancer ) , and KB ( oral-cavity cancer ) cell lines , with IC50 values of 2.7-4.5 microg/ml . Bauhinol B ( 2 ) is cytotoxic against NCI-H187 ( IC50 = 1.1 microg/ml ) and BC ( IC50 = 9.7 microg/ml ) cell lines , but inactive toward the KB cell line ( at 20 microg/ml ) . Compound 2 also is mildly antifungal towards Candia albicans ( IC50 = 28.9 microg/ml ) . Bibenzyl 6 is active against NCI-H187 ( IC50 = 14.1 microg/ml ) and BC ( IC50 = 4.0 microg/ml ) cells , but inactive ( at 20 microg/ml ) toward the KB cell line . Compounds 1 , 2 , and 6 show mild antimycobacterial activities , with MIC values of 25-50 microg/ml , but are inactive at 20 microg/ml against the P04264 malarial parasite strain ( Plasmodium falciparum ) . While bauhinol A ( 1 ) is inactive against cyclooxygenase 1 ( P23219 ) and cyclooxygenase 2 ( P35354 ) , compounds 2 and 6 inhibit both P23219 and P35354 , with IC50 values comparable to those of the standard drug , aspirin ( Table 3 ) . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Cloning , expression , and regulation of rabbit cyclooxygenase-2 in renal medullary interstitial cells . Prostaglandin synthesis requires cyclooxygenase-1 ( P23219 ) or -2 ( P35354 ) , which mediate the conversion of arachidonate to prostaglandin H2 . P23219 is the predominant constitutive isoform , whereas P35354 expression is typically low . In the present studies we cloned rabbit P35354 and determined its distribution in unstimulated tissues . Screening rabbit eye and uterine libraries yielded two cDNAs containing identical inserts with a 1,812-nucleotide open-reading frame . This encoded a 604-amino acid polypeptide , 90 % identical to human , rat , and mouse P35354 . Expression of the rabbit P35354 in P29320 -293 cells enhanced prostanoid synthesis . Constitutive P35354 mRNA expression was highest in kidney and urinary bladder . P35354 expression was primarily in renal outer medullary interstitial cells with cortical expression in macula densa . In cultured medullary interstitial cells , P35354 mRNA predominated , with little P23219 expression . Interstitial cell P35354 mRNA but not P23219 was induced by phorbol ester and epidermal growth factor but suppressed by dexamethasone . Phorbol ester also upregulated immunoreactive P35354 . Constitutive P35354 in these tissues has important implications for side effects of P35354 -selective inhibitors . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Estrogen response element-dependent regulation of transcriptional activation of estrogen receptors alpha and beta by coactivators and corepressors . One mechanism by which ligand-activated estrogen receptors alpha and beta ( ERalpha and ERbeta ) stimulate gene transcription is through direct ER interaction with specific DNA sequences , estrogen response elements ( EREs ) . ERE-bound ER recruits coactivators that stimulate gene transcription . Binding of ER to natural and synthetic EREs with different nucleotide sequences alters ER binding affinity , conformation , and transcriptional activity , indicating that the ERE sequence is an allosteric effector of ER action . Here we tested the hypothesis that alterations in ER conformation induced by binding to different ERE sequences modulates ER interaction with coactivators and corepressors . CHO- P04264 cells transfected with ERalpha or ERbeta show ERE sequence-dependent differences in the functional interaction of ERalpha and ERbeta with coactivators steroid receptor coactivator 1 ( Q15788 ) , P12931 -2 ( glucocorticoid receptor interacting protein 1 ( GRIP1 ) ) , Q9Y6Q9 amplified in breast cancer 1 ( Q9Y6Q9 ) and Q9Y6Q9 , cyclic AMP binding protein ( CBP ) , and steroid receptor RNA activator ( SRA ) , corepressors nuclear receptor co-repressor ( NCoR ) and silencing mediator for retinoid and thyroid hormone receptors ( Q9Y618 ) , and secondary coactivators coactivator associated arginine methyltransferase 1 ( Q86X55 ) and protein arginine methyltransferase 1 ( Q99873 ) . We note both ligand-independent as well estradiol- and 4-hydroxytamoxifen-dependent differences in ER-coregulator activity . In vitro ER-ERE binding assays using receptor interaction domains of these coregulators failed to recapitulate the cell-based results , substantiating the importance of the full-length proteins in regulating ER activity . These data demonstrated that the ERE sequence impacts estradiol-and 4-hydroxytamoxifen-occupied ERalpha and ERbeta interaction with coregulators as measured by transcriptional activity in mammalian cells . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Co-expression of the human cannabinoid receptor coding region splice variants ( hCB₁ ) affects the function of hCB₁ receptor complexes . The human type 1 cannabinoid ( hCB1 ) receptor is expressed at high levels in the central nervous system . mRNA variants of the coding region of this receptor , human cannabinoid hCB1a and hCB1b receptors , have been identified , their biological function remains unclear . The present study demonstrated that the three human cannabinoid hCB1 coding region variants are expressed in the human and monkey ( Macaca fascicularis ) brain . Western blot analyses of homogenates from different regions of the monkey brain demonstrated that proteins with the expected molecular weights of the cannabinoid P21554 , CB1a and CB1b receptors were co-expressed throughout the brain . Given the co-localization of these receptors , we hypothesized that physical interactions between the three splice variants may affect cannabinoid pharmacology . The human cannabinoid hCB1 , hCB1a , and hCB1b receptors formed homodimers and heterodimers , as determined by BRET in transiently transfected P29320 293A cells . We found that the co-expression of the human cannabinoid hCB1 and each of the splice variants increased cell surface expression of the human cannabinoid hCB1 receptor and increased Gi/o-dependent P29323 phosphorylation in response to cannabinoid agonists . Therefore , the human cannabinoid hCB1 coding region splice variants play an important physiological role in the activity of the endocannabinoid system . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . Expression of human (beta)3-adrenergic receptor induces adipocyte-like features in CHO/ P04264 fibroblasts . It is reported here that CHO/ P04264 cells stably transfected with the human (beta)3 AR gene ( CHO/ P04264 -(beta)3 ) , grown in the presence of differentiation-stimulating agents accumulate triglycerides . This lipid formation is mediated through the (beta)3 AR , since non-transfected CHO/ P04264 cells , or cells expressing the human (beta)2 AR , accumulate no significant amount of lipids when grown in supplemented medium . Moreover , lipid production can be inhibited significantly by the ( beta ) AR antagonist bupranolol . CHO/ P04264 cells expressing the W64R polymorphism ( DB00150 to DB00125 polymorphism at position 64 of the human (beta)3 AR ) , which has been associated with morbid obesity , show increased lipid accumulation as compared to CHO/ P04264 cells expressing the wild-type (beta)3 AR . Semi-quantitative RT-PCR experiments reveal that a major gene regulating adipocyte differentiation , peroxisome-proliferator-activated-receptor ( gamma ) ( Q07869 (gamma) ) , is expressed in CHO/ P04264 cells . Concomitantly with the formation of lipid droplets , the expression of Q07869 (gamma) mRNA is increased in CHO/ P04264 -(beta)3 cells , but not in non-transfected CHO/ P04264 cells . We furthermore detected constitutive expression of another adipocyte-associated protein : hormone sensitive lipase , while leptin or uncoupling protein-1 transcripts were not expressed . These data suggest that the frequently used CHO/ P04264 fibroblasts display several preadipocyte-like features , and that the sole expression of the (beta)3 AR modifies the expression of Q07869 (gamma) mRNA in these cells , and induces lipid formation under certain culture conditions . Curcumin inhibits the metastasis of P04264 papillary thyroid cancer cells via modulating P12830 and matrix metalloproteinase-9 expression . The anti-metastatic effect of curcumin on papillary thyroid cancer P04264 cells and its underlying mechanisms were investigated . Curcumin at 12.5 , 25 and 50 μM promoted mesenchymal-epithelial transition and decreased the migration rate of P04264 cells by 24-87 % . Its mechanism may involve the up-regulation of P12830 expression levels and down-regulation of the activity and expression of matrix metalloproteinase-9 . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Intake of vitamin P04264 and K2 and risk of hip fractures : The Hordaland Health Study . BACKGROUND : Evidence of the effect of vitamin K on bone health is conflicting . The aim was to investigate the association between intake of vitamins P04264 and K2 and subsequent risk of hip fracture in a general population sample , as well as potential effect modification by apolipoprotein E gene ( P02649 ) status by presence of the E4 allele . METHODS : 1238 men and 1569 women 71-75 years of age were included in the community-based Hordaland Health Study 1997-1999 in Western Norway . Information on hip fracture was obtained from hospitalizations in the region from enrolment until 31 December 2009 . Information on intake of vitamins P04264 and K2 collected at baseline was used as potential predictors of hip fracture in Cox proportional hazards regression analyses . RESULTS : Participants in the lowest compared to the highest quartile of vitamin P04264 intake had increased risk of suffering a hip fracture ( hazard ratio (HR)=1.57 [ 95 % CI 1.09 , 2.26 ] ) . Vitamin K2 intake was not associated with hip fracture . Presence of APOE4-allele did not increase the risk of hip fracture , nor was there any effect modification with vitamin P04264 in relation to risk of hip fracture . CONCLUSIONS : A low intake of vitamin P04264 , but not K2 , was associated with an increased risk of hip fractures . Titration of KATP channel expression in mammalian cells utilizing recombinant baculovirus transduction . A variety of transfection approaches have been used to deliver plasmids encoding ion channel genes into cells . We have used the baculovirus transduction system , BacMam , to demonstrate transient expression of multi-subunit KATP channels in CHO- P04264 and P29320 -293 EBNA cells using sulfonylurea receptor 1 ( Q09428 ) , SUR2A , SUR2B , and P55040 6.2 genes . [ 3H ] -glyburide binding , patch clamp , and DiBAC4(3) measurements of membrane potential changes were used to monitor channel expression . BacMam delivery of each Q09428 isoform with KIR6.2 was demonstrated based on its pharmacological profiles . Expression levels of Q09428 and KIR6.2 were titrated by varying the viral concentration or time of virus addition , with functional activity measured in as little as 4-6 hours posttransduction . Further increases in BacMam virus induced sufficient KATP expression to dominate membrane potential without pharmacological opening of the channel . Independently altering treatment with virus containing either the Q09428 or KIR6.2 gene revealed interactions among subunits during formation of functional channels in the plasma membrane . This study demonstrates the utility and versatility of BacMam as a valuable gene delivery tool for the study of ion channel function . A role for iron in Wnt signalling . There is an emerging body of evidence implicating iron in carcinogenesis and in particular colorectal cancer , but whether this involves Wnt signalling , a major oncogenic signalling pathway has not been studied . We aimed to determine the effect of iron loading on Wnt signalling using mutant P25054 ( Caco-2 and SW480 ) and wild-type P25054 ( P29320 -293 and human primary fibroblasts ) containing cell lines . Elevating cellular iron levels in Caco-2 and SW480 cells caused increased Wnt signalling as indicated by increased TOPFLASH reporter activity , increased mRNA expression of two known targets , c-myc and Nkd1 , and increased cellular proliferation . In contrast wild-type P25054 and beta-catenin-containing lines , P29320 293 and human primary fibroblasts were not responsive to iron loading . This was verified in SW480 cells that no longer induced iron-mediated Wnt signalling when transfected with wild-type P25054 . The cell line LS174T , wild type for P25054 but mutant for beta-catenin , was also responsive suggesting that the role of iron is to regulate beta-catenin . Furthermore , we show that P12830 status has no influence on iron-mediated Wnt signalling . We thus speculate that excess iron could exacerbate tumorigenesis in the background of P25054 loss , a common finding in cancers . Rat peroxisome proliferator-activated receptors and brown adipose tissue function during cold acclimatization . Brown adipose tissue ( Q14032 ) hyperplasia is a fundamental physiological response to cold ; it involves an acute phase of mitotic cell growth followed by a prolonged differentiation phase . Peroxisome proliferator-activated receptors ( PPARs ) are key regulators of fatty acid metabolism and adipocyte differentiation and may therefore mediate important metabolic changes during non-shivering thermogenesis . In the present study we have investigated Q07869 mRNA expression in relation to peroxisome proliferation in rat Q14032 during cold acclimatization . By immunoelectron microscopy we show that the number of peroxisomes per cytoplasmic volume and acyl- DB01992 oxidase immunolabeling density remained constant ( thus increasing in parallel with tissue mass and cell number ) during the initial proliferative phase and the acute thermogenic response but increased after 14 days of cold exposure , correlating with terminal differentiation of Q14032 . A pronounced decrease in Q14032 PPARalpha and PPARgamma mRNA levels was found within hours of exposure to cold , which was reversed after 14 days , suggesting a role for either or both of these subtypes in the proliferation and induction of peroxisomes and peroxisomal beta-oxidation enzymes . In contrast , PPARdelta mRNA levels increased progressively during cold exposure . Transactivation assays in HIB 1B and P29320 -293 cells demonstrated an adrenergic stimulation of peroxisome proliferator response element reporter activity via Q07869 , establishing a role for these nuclear receptors in hormonal regulation of gene transcription in Q14032 .

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interacts_with DB04817?

multiple_choice

8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . Effect of active smoking on the human bronchial epithelium transcriptome . BACKGROUND : Lung cancer is the most common cause of cancer-related deaths . Tobacco smoke exposure is the strongest aetiological factor associated with lung cancer . In this study , using serial analysis of gene expression ( Q9NXZ1 ) , we comprehensively examined the effect of active smoking by comparing the transcriptomes of clinical specimens obtained from current , former and never smokers , and identified genes showing both reversible and irreversible expression changes upon smoking cessation . RESULTS : Twenty-four Q9NXZ1 profiles of the bronchial epithelium of eight current , twelve former and four never smokers were generated and analyzed . In total , 3,111,471 Q9NXZ1 tags representing over 110 thousand potentially unique transcripts were generated , comprising the largest human Q9NXZ1 study to date . We identified 1,733 constitutively expressed genes in current , former and never smoker transcriptomes . We have also identified both reversible and irreversible gene expression changes upon cessation of smoking ; reversible changes were frequently associated with either xenobiotic metabolism , nucleotide metabolism or mucus secretion . Increased expression of Q07654 , O75952 , and Q5MY95 were found to be reversible upon smoking cessation . Expression of P49841 , which regulates P35354 expression , was irreversibly decreased . P98088 expression was only partially reversed . Validation of select genes was performed using quantitative RT-PCR on a secondary cohort of nine current smokers , seven former smokers and six never smokers . CONCLUSION : Expression levels of some of the genes related to tobacco smoking return to levels similar to never smokers upon cessation of smoking , while expression of others appears to be permanently altered despite prolonged smoking cessation . These irreversible changes may account for the persistent lung cancer risk despite smoking cessation . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . P05362 -independent lymphocyte transmigration across high endothelium : differential up-regulation by interferon gamma , tumor necrosis factor-alpha and interleukin 1 beta . The adhesion of lymphocytes to cytokine-treated high endothelium was studied using cultured high endothelial cells ( O14777 ) . Pretreatment of the O14777 layer with a variety of cytokines caused up-regulation of lymphocyte adhesion with the effects ordered interferon gamma ( P01579 ) greater than tumor necrosis factor-alpha ( P01375 ) greater than or equal to interleukin 1 beta ( IL 1 beta ) . Increased lymphocyte adhesion was found to be independent of P05362 as expression by O14777 was not increased by cytokines and antibodies against P05362 did not block adhesion . The peptide CS1 and anti-beta 1 integrin subunit antibodies , however , caused partial inhibition of lymphocyte adhesion thus indicating a role for fibronectin on O14777 and alpha 4 beta 1 on lymphocytes . Study of the kinetics of lymphocyte adhesion showed that the effects of P01579 and P01375 were persistent and remained detectable 2.5 h after removal of the cytokines whereas the effects of IL 1 beta were transient and were not sustained beyond 1 h . All of the cytokines used caused transient increases in the number of surface-bound lymphocytes with P01579 greater than P01375 greater than or equal to IL 1 beta , however , the most dramatic effect was on the transmigration of lymphocytes across the O14777 . Both P01579 and P01375 caused sustained increased transmigration with P01579 having the greater effect . IL 1 beta had little effect on transmigration . This model demonstrates that the binding and transmigration of lymphocytes across O14777 can be differentially regulated by the actions of individual cytokines . These results support the concept that locally produced cytokines regulate O14777 function within the lymph node . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . The role of celecoxib in Rad51 expression and cell survival affected by gefitinib in human non-small cell lung cancer cells . Celecoxib ( DB00482 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor and gefitinib ( DB00317 (R) , ZD1839 ) is a selective epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor for human non-small cell lung cancer ( NSCLC ) . The addition of celecoxib to gefitinib to prolong the survival of patients with NSCLC still remains controversial and needs to be investigated . The Rad51 protein is essential for homologous recombination repair , and is overexpressed in chemo- or radioresistant carcinomas . In this study , we characterize the role of celecoxib in the cytotoxicity , P27361 /2 activation and Rad51 expression affected by gefitinib in NSCLC cells . We show that celecoxib can enhance the cytotoxicity induced by gefitinib in NSCLC cells . Treatment with celecoxib alone has no effect on the P27361 /2 activation , Rad51 mRNA and protein levels , however , combined treatment with gefitinib results in a significant reduction of phospho- P27361 /2 and Rad51 protein levels , and triggers the degradation of Rad51 via a 26S proteasome-dependent pathway . Expression of constitutively active Q02750 /2 vectors ( Q02750 /2-CA ) significantly rescues the decreased P27361 /2 activity , and restores Rad51 protein levels and cell survival under co-treatment with gefitinib and celecoxib . Furthermore , blocking P27361 /2 activation by U0126 ( Q02750 /2 inhibitor ) and knocking down Rad51 expression by transfection with small interfering RNA of Rad51 can enhance the cytotoxicity of celecoxib . Fucosylated glycoproteomic approach to identify a complement component 9 associated with squamous cell lung cancer ( SQLC ) . Human lung cancer is a major cause of cancer mortality worldwide . Understanding the pathophysiological features and the development of novel biomarkers for diagnosis as well as treatment are major tasks . In the present study , sera from ten SQLC patients and healthy control ( O14777 ) were collected and pooled , respectively . The pooled sera were depleted via an immunoaffinity method and further subjected to fucosylation enrichment . Enriched fucosylated glycoproteins were resolved by SDS-PAGE and subsequently analyzed by LC- P19957 -MS/MS . From comparative proteomic analysis , we selected the P02748 protein . P02748 protein levels were validated by Western blot , protein arrays and the fucosylation levels of P02748 by hybrid lectin ELISA ( P08246 ) in the sera of 120 O14777 and 118 SQLC patients . The P02748 protein level was 6.4-fold higher in SQLC patients compared to O14777 , as determined by Western blot analysis . The results were concurrently confirmed by a protein array that showed a P02748 level significantly higher in SQLC patients , as compared to O14777 , with a sensitivity of 53 % and a specificity of 89 % . P02748 fucosylation levels were significantly higher in SQLC patients compared to O14777 ( p < 0.05 ) when tested by P08246 . These findings suggest that P02748 and fucosylated form could serve as a useful marker for SQLC . Anti-allergic effects of nilotinib on mast cell-mediated anaphylaxis like reactions . DB04868 is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of P11274 - P00519 -positive chronic myelogenous leukemia . However , its effect on mast cell-mediated anaphylactic reaction is still not known . The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action . DB04868 administration prevented systemic anaphylaxis in mice , mediated by compound 48/80 , in a dose- and time-dependent manner . Also , nilotinib significantly inhibited ( P < 0.05 ) allergic paw edema in rats . Furthermore , nilotinib significantly decreased ( P < 0.05 ) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner . In addition , nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin . Moreover , nilotinib attenuated the secretion of pro-inflammatory cytokine , tumor necrosis factor ( P01375 ) -α expression in the rat peritoneal mast cells . These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent . A comparative study of the antipyretic effects of indomethacin and dipyrone in rats . OBJECTIVE : Compare the antipyretic effects of dipyrone and indomethacin . MATERIALS AND METHODS : Fever was induced in rats by i. v. LPS or i . c . v. interleukins ( IL ) , prostaglandins ( PG ) , arachidonic acid ( AA ) , pre-formed pyrogenic factor ( PFPF ) , tumour necrosis factor-alpha ( P01375 ) or corticotrophin releasing hormone ( P06850 ) . DB04817 and indomethacin were administered i.p. , arginine vasopressin V1-receptor antagonist , d(CH2)5 DB00135 (Me)AVP , into the ventral septal area . Cyclooxygenase ( P23219 /-2 ) blocking activity was assessed in transfected COS-7 cells . P06850 release from isolated hypothalami was determined by ELISA . RESULTS : Indomethacin or dipyrone reduced LPS , IL-1beta , P05231 or P01375 induced fever and P06850 release from rat hypothalamus . Only dipyrone inhibited P10145 , PFPF or PGF2alpha fever . Only indomethacin inhibited fever induced by AA or IL-1beta , plus AA . Neither antipyretic affected fever caused by DB00917 or P06850 . d(CH2)5Tyr(Me)AVP only blocked antipyresis induced by indomethacin . DB04817 at a very high concentration ( 10 mM ) inhibited only P23219 , while indomethacin ( 0.1 microM ) blocked P23219 and P35354 in COS-7 cells . CONCLUSION : The antipyretic effect of dipyrone differs from that of indomethacin in that it does not depend on AVP release or inhibition of PG synthesis . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Celecoxib blocks cardiac Kv1.5 , Kv4.3 and Kv7.1 ( P51787 ) channels : effects on cardiac action potentials . Celecoxib is a P35354 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets . The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels ( Kv ) involved on cardiac repolarization Kv1.5 ( I(Kur) ) , Kv4.3+ Q9NS61 ( I(to1) ) and Kv7.1+ P15382 ( I(Ks) ) and to compare with another P35354 inhibitor , rofecoxib . Currents were recorded in transfected mammalian cells by whole-cell patch-clamp . Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent , except on Kv4.3+ Q9NS61 channels . Kv1.5 block increased in the voltage range of channel activation , decreasing at potentials positive to 0 mV . The drug modified the activation curve of the channels that became biphasic . Block was frequency-dependent , increasing at fastest frequencies . Celecoxib effects were not altered by DB08837 (out) in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with DB08837 (in) , indicating that celecoxib acts from the cytosolic side . We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells , where Kv1.5 are the main contributors ( IC(50)≈ 7 μM ) . Finally , we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes , suggesting that Kv block induced by celecoxib may be of clinical relevance . N-glucuronidation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) and N_hydroxy-PhIP by specific human UDP-glucuronosyltransferases . Glucuronidation is a major metabolic pathway in the biotransformation of many xenobiotics . Recent studies have shown that in humans , UDP-glucuronosyltransferase ( P78381 ) -mediated glucuronidation plays a critical role in the detoxification of food-borne carcinogenic heterocyclic amines . 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , the most abundant carcinogenic heterocyclic amine found in well-cooked meats , has been shown to be extensively glucuronidated in humans . To determine which P78381 isozymes are involved in the biotransformation of PhIP and the cytochrome P4501A2-mediated reactive intermediate N-hydroxy-PhIP , microsomes expressing human P22309 , -1A4 , -1A6 or -1A9 were incubated with PhIP and N-hydroxy-PhIP and the reaction products analyzed by HPLC and P19957 -MS . Incubations containing N-hydroxy-PhIP and P22309 expressing microsomes , with an apparent Km of 4.58 microM and a Vmax of 4.18 pmol/min/mg protein , had the highest capacity to convert N-hydroxy-PhIP to N-hydroxy-PhIP-N2-glucuronide . Microsomes expressing O60656 produced N-hydroxy-PhIP-N3-glucuronide at the highest rate with an apparent Km and Vmax of 3.73 microM and 4.07 pmol/min/mg , respectively . A third previously undefined glucuronide accounted for 31 % of the total glucuronides formed from the P22310 expressing microsomes . No glucuronide conjugates were detected from microsomes expressing P19224 . Incubations containing PhIP as substrate formed direct PhIP-glucuronides in microsomes expressing P22309 , P22310 and O60656 but at levels averaging 53-fold lower than when N-hydroxy-PhIP was used as the substrate . Knowing the glucuronidation capacity of the specific P78381 isozymes involved in PhIP and N-hydroxy-PhIP glucuronidation should help in determining the individual susceptibility to the potential cancer risk from exposure to PhIP . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . Ectopic DB01285 syndrome caused by bronchial carcinoid tumor indistinguishable from Cushing 's disease . A 75-year-old woman was admitted to our hospital because of a poor glycemic control . She was found to have Cushingoid feature and dynamic endocrine tests showed elevated plasma DB01285 and cortisol levels , lack of their circadian rhythm , non-suppressibility to high-dose dexamethasone , responsiveness to P06850 , but not to DB00035 , and suppression to octreotide . Pituitary Q9BWK5 showed an equivocal small lesion . CT scan of the chest showed two nodular lesions in the right lung ( S5 , S7 ) , while a mild uptake was noted only in S5 lesion by DB09150 -PET , but positive uptake was only in S7 lesion by somatostatin receptor scintigraphy ( SRS ) . Inferior petrosal sinus sampling revealed a gradient of plasma DB01285 after P06850 stimulation , consistent with the diagnosis of Cushing ' s disease . She underwent middle and inferior lobectomy of the right lung . The resected tumor in S7 was consistent with the diagnosis of a bronchial carcinoid tumor with positive DB01285 immunoreactivity , while that of S5 was cryptococcal granuloma . RT-PCR revealed abundant expressions of P01189 and SSTR ( -1 , -2 , -5 ) , but not of P34998 and P47901 . Postoperatively , abnormal endocrine data were normalized along with improvement of hypertension and diabetes . This was a diagnostic challenging case with ectopic DB01285 syndrome indistinguishable from Cushing ' s disease by various endocrine and imaging tests , among which SRS successfully localized the tumor responsible for ectopic DB01285 secretion . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . Surfactin exhibits neuroprotective effects by inhibiting amyloid β-mediated microglial activation . Microglial-mediated neuroinflammation and neurotoxicity contribute to the pathogenesis of neurodegenerative diseases including Alzheimer 's disease ; therefore , control of microglial activation and subsequent suppression of neurotoxic pro-inflammatory molecules could provide a potential therapeutic approach for the treatment of such diseases . In this study , we investigated the effects of surfactin , a surfactant from Bacillus subtilis , on oligomeric amyloid β ( Aβ ) -induced microglial activation and neurotoxicity . Surfactin significantly suppressed expression of P14780 , P35228 and P35354 , as well as production of ROS , NO , DB00917 , P01375 -α , IL-1β , P05231 and P13500 in Aβ-stimulated BV-2 microglial cells . Moreover , surfactin markedly inhibited Aβ-induced nuclear translocation and activation of NF-κB as well as phosphorylation of JNK and p38 MAPK . Furthermore , surfactin protected hippocampal HT22 cells from indirect neuronal toxicity mediated by Aβ-treated microglial cells , but had no effect on Aβ-induced direct toxicity to HT22 cells . These results suggest that surfactin impairs the Aβ-induced inflammatory response of microglial cells and confers protection against indirect neurotoxicity to hippocampal cells . Our findings indicate that surfactin may have therapeutic potential for ameliorating Alzheimer 's disease as well as other neurodegenerative disorders which involve neuroinflammation . Synthesis and biological evaluation of novel pyrrolidine-2,5-dione derivatives as potential antidepressant agents . Part 1 . A series of 3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by (1)H NMR , (13)C NMR and P19957 -HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5- Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists . Novel bioactive metabolites of dipyrone ( metamizol ) . DB04817 is a common antipyretic drug and the most popular non-opioid analgesic in many countries . In spite of its long and widespread use , molecular details of its fate in the body are not fully known . We administered dipyrone orally to mice . Two unknown metabolites were found , viz. the arachidonoyl amides of the known major dipyrone metabolites , 4-methylaminoantipyrine ( 2 ) and 4-aminoantipyrine ( 3 ) . They were identified by P19957 -LC-MS/MS after extraction from the CNS , and comparison with reference substances prepared synthetically . The arachidonoyl amides were positively tested for cannabis receptor binding ( CB(1) and CB(2) ) and cyclooxygenase inhibition ( P23219 and P35354 in tissues and as isolated enzymes ) , suggesting that the endogenous cannabinoid system may play a role in the effects of dipyrone against pain . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Therapeutic targeting of CPT-11 induced diarrhea : a case for prophylaxis . CPT-11 ( irinotecan ) , a P11387 inhibitor is one of the main treatments for colorectal cancer . The main dose limiting toxicities are neutropenia and late onset diarrhea . Though neutropenia is manageable , CPT-11 induced diarrhea is frequently severe , resulting in hospitalizations , dose reductions or omissions leading to ineffective treatment administration . Many potential agents have been tested in preclinical and clinical studies to prevent or ameliorate CPT-11 induced late onset diarrhea . It is predicted that prophylaxis of CPT-11 induced diarrhea will reduce sub-therapeutic dosing as well as hospitalizations and will eventually lead to dose escalations resulting in better response rates . This article reviews various experimental agents and strategies employed to prevent this debilitating toxicity . Covered topics include schedule/dose modification , intestinal alkalization , structural/chemical modification , genetic testing , anti-diarrheal therapies , transporter ( P08183 , Q92887 , Q96JK2 ) inhibitors , enzyme ( β-glucuronidase , P22309 , P08684 , carboxylesterase , P35354 ) inducers and inhibitors , probiotics , antibiotics , adsorbing agents , cytokine and growth factor activators and inhibitors and other miscellaneous agents . Increased constitutive activity of P31749 /Akt in tamoxifen resistant breast cancer MCF-7 cells . The tamoxifen-resistant ( TAM-R ) MCF-7 breast cancer cell line has been used as a model to identify the signalling pathways that enable resistant cancer cells to grow independently of steroid hormones . In TAM-R cells , peptide growth factor signalling pathways appear to be important in modified cell behaviour , growth and survival . The P19957 kinase signalling components Akt1 and Akt2 are expressed at similar levels by both parental wild-type MCF-7 and TAM-R cells , but Akt1 phosphorylation is significantly increased in TAM-R cells grown under basal conditions . High levels of basal Akt , GSK3 alpha / beta and p70S6 kinase phosphorylation are all inhibited by the P19957 kinase inhibitor , LY 294002 . The ligands for the P00533 /erbB1 receptor , P01133 ( epidermal growth factor ) and TGF alpha ( transforming growth factor- alpha ) demonstrate an increased ability to activate Akt in TAM-R compared with parental MCF-7 cells and it is proposed that the preferred autocrine or paracrine activation of Akt occurs via the erbB heterodimer P00533 /erbB2 in TAM-R cells . Akt phosphorylation is reduced by gefitinib ( " DB00317 " /ZD1839 ) . The results suggest that the P19957 kinase pathway plays a role in proliferation of TAM-R cells and is important in the increased P01133 induced membrane ruffling detected in the resistant cells . Increased Akt1 activation may contribute to the aggressive phenotype of tamoxifen resistant ER ( oestrogen receptor ) positive breast cancers . P14210 induces anoikis resistance by up-regulation of cyclooxygenase-2 expression in uterine endometrial cancer cells . P35354 ( P35354 ) has been implicated in the promotion of carcinogenesis . Although the role of P35354 in endometrial cancer remains unclear , recent experiments suggest that P35354 antagonizes cell apoptosis , increases the invasiveness of malignant cells , and promotes angiogenesis . P14210 ( P14210 ) is a mesenchymal-derived cytokine and the interaction between P14210 and its tyrosine kinase receptor , c- DB00134 proto-oncogene , is associated with tumor progression and metastasis . To investigate the molecular mechanism of P14210 -induced anoikis resistance , we analyzed the signal transduction and P35354 expression in endometrial cancer cells . Here , we show i ) the expression of P35354 protein significantly increased in a dose-dependent manner after P14210 stimulation in endometrial cancer cell lines ( O14777 -IB and RL95-2 ) , reaching 200-270 % stimulation at the highest doses of P14210 tested ( 40 ng/ml ) ; ii ) flow cytometry and TUNEL analyses revealed that P14210 significantly inhibited anoikis of RL95-2 cells ; iii ) phosphatidylinositol 3-kinase ( PI3K ) inhibitor ( LY294002 ) , but not mitogen-activated protein kinase/ P29323 kinase ( MEK ) inhibitor ( PD98059 ) , specifically blocked P14210 -mediated anoikis resistance in RL95-2 cells ; and iv ) P35354 inhibitor , Meloxicam , abrogated P14210 -mediated anoikis resistance . Our data suggest that P14210 induces anoikis resistance in endometrial cancer cells possibly through PI3K/Akt pathway-dependent up-regulation of P35354 expression . Evaluation of the endogenous cannabinoid system in mediating the behavioral effects of dipyrone ( metamizol ) in mice . DB04817 is a common nonopioid analgesic and antipyretic , which , in many countries , is available over the counter and is more widely used than paracetamol or aspirin . However , the exact mechanisms by which dipyrone acts remain inconclusive . Two novel arachidonoyl-conjugated metabolites are formed in mice following the administration of dipyrone that are dependent on the activity of fatty acid amide hydrolase ( FAAH ) , which also represents the major catabolic enzyme of the endogenous cannabinoid ligand anandamide . These arachidonoyl metabolites not only inhibit cyclooxygenase ( P23219 / P35354 ) but also bind to cannabinoid receptors at low micromolar concentrations . The relative contributions of cannabinoid receptors and FAAH in the overall behavioral response to dipyrone remain untested . Accordingly , the two primary objectives of the present study were to determine whether the behavioral effects of dipyrone would ( a ) be blocked by cannabinoid receptor antagonists and ( b ) occur in FAAH mice . Here , we report that thermal antinociceptive , hypothermic , and locomotor suppressive actions of dipyrone are mediated by a noncannabinoid receptor mechanism of action and occurred after acute or repeated administration irrespective of FAAH . These findings indicate that FAAH-dependent arachidonoyl metabolites and cannabinoid receptors are not requisites by which dipyrone exerts these pharmacological effects under noninflammatory conditions . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients .

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MH_train_1074

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interacts_with DB01017?

multiple_choice

Release of mediators of systemic inflammatory response syndrome in the course of a severe delayed hemolytic transfusion reaction caused by anti-D . BACKGROUND : In vitro studies suggest that mediators of systemic inflammatory response syndrome are generated in the course of hemolytic transfusion reactions . Evidence for the in vivo significance of these findings is given by the present clinical and laboratory analysis of a severe delayed hemolytic transfusion reaction ( P10275 ) . CASE REPORT : A 67-year-old patient ( blood group O , D-negative ) with a negative pretransfusion antibody screen received a massive transfusion because of arterial bleeding ( Day 1 ) . The transfusion of group O , D-positive red cell concentrates was unavoidable because of limited supplies . At Day 10 , the patient developed a P10275 with symptoms of septic-toxic syndrome and signs of hemolysis ; he received an exchange transfusion . Serologic markers , as well as proinflammatory and anti-inflammatory mediators , were monitored at the onset of the P10275 and during the exchange transfusion . RESULTS : At Day 10 , the direct antiglobulin test was positive ; anti-D was present , most likely as the result of an anamnestic immune response . Interleukin ( IL ) -1 was not detectable ; all other mediators monitored were elevated : IL-1 receptor antagonist , tumor necrosis factor , P05231 , P10145 , P22301 , neopterin , elastase , C3a-desArg , P02741 , and fibrinogen . Most of the values declined during the exchange transfusion , which was followed by an improvement of the clinical presentation . CONCLUSIONS : Mediators of systemic inflammatory response syndrome were released in the course of a P10275 caused by anti-D . Severe clinical symptoms could be treated successfully by exchange transfusion . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . [ Anti-inflammatory effect of Urtica dioica folia extract in comparison to caffeic malic acid ] . Urtica dioica extract is a traditionary used adjuvant therapeutic in rheumatoid arthritis . The antiphlogistic effects of the urtica dioica folia extract P22304 23 ( Extractum Urticae dioicae foliorum ) and the main phenolic ingredient caffeic malic acid were tested concerning the inhibitory potential on biosynthesis of arachidonic acid metabolites in vitro . The caffeic malic acid was isolated from Urtica folia extract using gel exclusion- and high performance liquid chromatography and identified by mass spectroscopy and nuclear magnetic resonance . Concerning the P09917 products P22304 23 showed a partial inhibitory effect . The isolated phenolic acid inhibited the synthesis of the leukotriene B4 in a concentration dependent manner . The concentration for halfmaximal inhibition ( IC50 ) was 83 microns/ml in the used assay . P22304 23 showed a strong concentration dependent inhibition of the synthesis of cyclooxygenase derived reactions . The IC50 were 92 micrograms/ml for P22304 23 and 38 micrograms/ml for the caffeic malic acid . Calculating the content in P22304 23 the caffeic malic acid is a possible but not the only active ingredient of the plant extract in the tested assay systems . It is demonstrated that the phenolic component showed a different enzymatic target compared with P22304 23 . The antiphlogistic effects observed in vitro may give an explanation for the pharmacological and clinical effects of P22304 23 in therapie of rheumatoid diseases . DB01017 protects PC12 cells against DB01221 -induced injury via inhibiting P09917 activation . Recently , we have reported that minocycline , a semi-synthetic tetracycline with neuroprotective effects , inhibits the in vitro ischemic-like injury and P09917 ( 5- P28300 ) activation in PC12 cells . In the present study , we further determined whether minocycline protects PC12 cells from excitotoxicity via inhibiting 5- P28300 activation . We used N-methyl-d-aspartate ( DB01221 , 200 microM ) to induce early ( exposure for 6 h ) and delayed ( exposure for 6 h followed by 24 h recovery ) injuries . We found that DB01221 receptor antagonist ketamine , 5- P28300 inhibitor caffeic acid and minocycline concentration dependently attenuated DB01221 -induced early and delayed cell injuries ( viability reduction and cell death ) . However , only ketamine ( 1 microM ) inhibited DB01221 -evoked elevation of intracellular calcium . In addition , immunohistochemical analysis showed that DB01221 induced 5- P28300 translocation to the nuclear membrane after 1- to 6-h exposure which was confirmed by Western blotting , indicating that 5- P28300 was activated . DB01221 , caffeic acid and minocycline ( each at 1 microM ) inhibited 5- P28300 translocation after early injury . After delayed injury , PC12 cells were shrunk , and 5- P28300 was translocated to the nuclei and nuclear membrane ; ketamine , caffeic acid and minocycline inhibited both cell shrinking and 5- P28300 translocation . As a control , P16050 inhibitor baicalein showed a weak effect on cell viability and death , but no effect on 5- P28300 translocation . Therefore , we conclude that the protective effect of minocycline on DB01221 -induced injury is partly mediated by inhibiting 5- P28300 activation . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . DB01017 modulates cytokine and gene expression profiles in the brain after whole-body exposure to radiation . An effective countermeasure against radiation damage to normal tissues is urgently needed . The major goal of the present study was to determine if minocycline could modify the immunomodulatory effects of radiation on the brain . C57BL/6 mice were treated with minocycline intraperitoneally for 5 days beginning immediately before total-body exposure to 0 , 1 , 2 and 3 Gray ( Gy ) (60)Co γ-rays . Brains were collected on days 4 and 32 post-irradiation for cytokine and gene analyses . DB01017 treatment significantly increased the levels of interleukin ( IL ) -10 , P40933 and vascular endothelial growth factor ( P15692 ) in the brain on day 4 in one or more irradiated groups compared to radiation-alone ( p < 0.05 ) . P22301 is anti-inflammatory , P40933 can prevent apoptosis and P15692 is nuroprotective . On day 32 , the drug decreased IL-1β in the 2- Gy group ( p < 0.05 vs. 2-Gy alone ) ; this cytokine is implicated in immune-related central nervous system pathologies . Microarray analysis of brains on day 32 showed that while radiation increased expression of inflammatory genes such as Il1f10 , Il17 , Tnfrsf11b , Tnfsf12 , Il12b and Il1f8 , these were no longer up-regulated in the minocycline-treated groups . Similarly , the pro-apoptotic gene Bik and nitric oxide synthase producer ( Q8IVI9 ) were no longer up-regulated in the drug-treated groups . Pathway analysis based on gene data suggested that catenin-β1 and tumor suppressor-related transcription regulation were significantly activated by radiation and/or minocycline ( activation z-score > 2.0 ) . Overall , the data warrant further testing of minocycline as a potential neuroprotectant against radiation-induced damage . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . DB01017 accelerates hypoxia-inducible factor-1 alpha degradation and inhibits hypoxia-induced neovasculogenesis through prolyl hydroxylase , von Hippel-Lindau-dependent pathway . Hypoxia-mediated stress responses are important in tumor progression , especially when tumor growth causes the tumor to become deprived of its blood supply . The oxygen-labile transcription factor hypoxia-inducible factor-1 alpha ( HIF-1α ) plays a critical role in regulating hypoxia stress-related gene expression and is considered a novel therapeutic target . Lung adenocarcinoma cell lines were exposed to minocycline , followed by incubation at hypoxic condition for 3-6 h . Here , we show that minocycline , a second-generation tetracycline , can induce HIF-1α proteasomal degradation under hypoxia by increasing the expression prolyl hydroxylase-2 and HIF-1α/von Hippel-Lindau protein interaction , thereby overcoming hypoxia-induced HIF-1α stabilization . Neither repression of hypoxia-induced phosphatidylinositol-3 kinase/Akt/mammalian target of rapamycin pathway nor inhibition of Hsp90 was required for minocycline-induced HIF-1α degradation . The HIF-1α degradation-enhancing effect of minocycline was evident in both cancerous and primary cells . DB01017 -pretreated , hypoxia-conditioned cells showed a clear reduction in hypoxia response element reporter expression and amelioration of vascular endothelial growth factor C/D ( P49767 /D ) , matrix metalloproteinase 2 , and glucose transporter 1 expression . By decreasing P15692 secretion of cancerous cells , minocycline could suppress endothelial cell neovasculogenesis . These findings suggest a novel application of minocycline in the treatment of tumor angiogenesis as well as hypoxia-related diseases . Chemotactic and cytotoxic effects of minocycline on human retinal pigment epithelial cells . PURPOSE : To reveal the effects of minocycline , an anti-inflammatory and neuroprotective agent , on the viability and physiological properties of retinal pigment epithelial ( Q96AT9 ) cells and to compare the effects with those of triamcinolone acetonide . METHODS : The proliferation of human Q96AT9 cells in vitro was investigated with a bromodeoxyuridine immunoassay ; chemotaxis was examined with a Boyden chamber assay . Cell viability was determined by trypan blue exclusion . The gene expression of growth factors and P14780 was determined with real-time RT-PCR , and the secretion of P15692 was examined with ELISA . The phosphorylation of p38 MAPK and P27361 /2 proteins was determined with Western blot analysis . RESULTS : DB01017 at low concentrations ( 50 nM-20 microM ) stimulated chemotaxis and decreased the proliferation of Q96AT9 cells . DB01017 at high concentrations ( above 5 microM ) decreased the viability of Q96AT9 cells through the induction of cell necrosis . The chemotactic effect of minocycline was mediated by the stimulation of autocrine PDGF signaling and the activation of p38 MAPK . DB01017 promoted the expression of PDGF-B , P14210 , P15692 , and P14780 and increased the amounts of phosphorylated p38 and P27361 /2 proteins in Q96AT9 cells . DB00620 reduced PDGF-evoked chemotaxis and P15692 expression and secretion and had no significant effects on cell viability and proliferation . DB00620 did not reverse the effects of minocycline on cell proliferation , chemotaxis , or viability or the expression of P15692 . CONCLUSIONS : Low-dose minocycline induces the activation of Q96AT9 cells , as indicated by the activation of p38 and P27361 /2 and by enhanced chemotaxis mediated by autocrine PDGF signaling . High-dose minocycline induces Q96AT9 cell degeneration . DB01017 protects PC12 cells from ischemic-like injury and inhibits P09917 activation . DB01017 protects animals against cerebral ischemia by inhibiting inflammation . To determine whether minocycline protects PC12 cells from in vitro ischemic-like injury and affects pro-inflammatory P09917 activation , the cell viability and P09917 translocation to nuclear membrane were observed after oxygen-glucose deprivation ( OGD ) . We found that OGD reduced cell viability , which was attenuated by minocycline and P09917 inhibitor caffeic acid . 5-Lipoxygenase protein was detected in PC12 cells by immunohistochemical and Western blot analyses . OGD induced P09917 translocation to nuclear membranes , which was abolished by minocycline and caffeic acid . Thus , minocycline can protect PC12 cells from in vitro ischemic-like injury , and this effect may be partly related to the inhibition of P09917 activation . Levels of angiopoietins 1 and 2 in induced sputum supernatant in patients with P48444 . Pathological features of chronic obstructive pulmonary disease ( P48444 ) include lung vascular remodeling and angiogenesis . Q15389 ( Ang-1 ) , is an essential mediator of angiogenesis by establishing vascular integrity , whereas angiopoietin-2 ( Ang-2 ) acts as its natural inhibitor . We determined the levels of angiopoietins in sputum supernatants of patients with P48444 and investigated their possible association with mediators and cells involved in the inflammatory and remodeling process . Fifty-nine patients with P48444 , 25 healthy smokers and 20 healthy non-smokers were studied . All subjects underwent lung function tests , sputum induction for cell count identification and Ang-1 , Ang-2 , P15692 , TGF-β1 , P08253 , LTB4 , P10145 , albumin measurement in sputum supernatants . Airway vascular permeability ( AVP ) index was also assessed . Ang-2 levels were significantly higher in patients with P48444 compared to healthy smokers and healthy non-smokers [ median , interquartile ranges pg/ml , 267 ( 147-367 ) vs. 112 ( 67-171 ) and 98 ( 95-107 ) , respectively ; p < 0.001 ] . Regression analysis showed a significant association between Ang-2 levels and AVP index , P15692 , P10145 and P08253 levels in P48444 , the strongest being with P15692 . Our results indicate that induced sputum Ang-2 levels are higher in P48444 compared to healthy smokers and healthy non-smokers . Moreover , Ang-2 is associated with AVP , P10145 , P08253 , and P15692 , indicating a possible role for Ang-2 in the pathogenesis of the disease . DB01017 and tissue-type plasminogen activator for stroke : assessment of interaction potential . BACKGROUND AND PURPOSE : New treatment strategies for acute ischemic stroke must be evaluated in the context of effective reperfusion . DB01017 is a neuroprotective agent that inhibits proteolytic enzymes and therefore could potentially both inactivate the clot lysis effect and decrease the damaging effects of tissue-type plasminogen activator ( t-PA ) . This study aimed to determine the effect of minocycline on t-PA clot lysis and t-PA-induced hemorrhage formation after ischemia . METHODS : Fibrinolytic and amidolytic activities of t-PA were investigated in vitro over a range of clinically relevant minocycline concentrations . A suture occlusion model of 3-hour temporary cerebral ischemia in rats treated with t-PA and 2 different minocycline regimens was used . Blood-brain barrier basal lamina components , matrix metalloproteinases ( MMPs ) , hemorrhage formation , infarct size , edema , and behavior outcome were assessed . RESULTS : DB01017 did not affect t-PA fibrinolysis . However , minocycline treatment at 3 mg/kg IV decreased total protein expression of both P08253 ( P=0.0034 ) and P14780 ( P=0.001 for 92 kDa and P=0.0084 for 87 kDa ) . It also decreased the incidence of hemorrhage ( P=0.019 ) , improved neurologic outcome ( P=0.0001 for Bederson score and P=0.0391 for paw grasp test ) , and appeared to decrease mortality . MMP inhibition was associated with decreased degradation in collagen IV and laminin-alpha1 ( P=0.0001 ) . CONCLUSIONS : Combination treatment with minocycline is beneficial in t-PA-treated animals and does not compromise clot lysis . These results also suggest that neurovascular protection by minocycline after stroke may involve direct protection of the blood-brain barrier during thrombolysis with t-PA . Understanding the molecular mechanism of blood-brain barrier damage in an experimental model of Japanese encephalitis : correlation with minocycline administration as a therapeutic agent . The blood-brain barrier ( BBB ) serves to protect the central nervous system ( CNS ) from damage by exogenous molecules . Japanese encephalitis ( JE ) , caused by a neurotropic flavivirus , leads to inflammation in the CNS , neuronal death and also compromises the structural and functional integrity of the BBB . DB01017 , a semisynthetic tetracycline , has been found to be broadly protective in neurological disease models featuring inflammation and cell death and at present , is being evaluated in clinical trials . In the present study , we propose that the neuroprotective role of minocycline in experimental models of JE extends also to the protection of the BBB . Damage to the BBB was assessed by Evan 's blue dye exclusion test after minocycline treatment following Japanese encephalitis virus ( JEV ) infection . A breakdown of the BBB occurred in mice inoculated intravenously with JEV . This resulted in leakage of protein-bound Evan 's blue dye into the brain tissue . Semi-quantitative RT-PCR revealed an up-regulation of chemokine receptors and adhesion molecules following JEV infection . Immunostaining showed leukocyte and neutrophil infiltration following JEV infection . Intraperitoneal injection of minocycline , beginning 24h post-JEV infection , abrogated the effects by reducing BBB damage , decreasing expression of P35228 , Cox-2 , P15692 and also by reducing the elevated level of transcript of chemokine receptors and adhesion molecules in the brain . Matrix metalloproteinases ( MMPs ) are known to disrupt the BBB and minocycline was found to significantly decrease the activity of P14780 in brain tissue homogenates . Thus , minocycline , administered at a clinically relevant time , appears to maintain blood-brain barrier integrity following JEV infection . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Down-regulation of P12830 in mouse skin carcinoma cells enhances a migratory and invasive phenotype linked to matrix metalloproteinase-9 gelatinase expression . To assess the role of gelatinases in mouse skin tumor progression and their link to the expression of P12830 ( E-CD ) , the cell-cell adhesion protein , we used the highly metastatic squamous HaCa4 cell line and several HaCa4-derived clones obtained by transfection of the mouse E-CD cDNA . Expression of matrix metalloproteinase-9 ( P14780 ) mRNA and protein activity were present in E-CD ( - ) HaCa4 and control clones in culture , but they were strongly diminished in E-CD ( + ) clones ( E24 and E62 ) at subconfluence . To explore the suppressive effect of the cell-cell contacts mediated by E-CD on P14780 expression , we introduced a plasmid encoding mouse E-CD antisense cDNA into the E24 cell clone . The transfectant P1-clones obtained with reduced or absent E-CD expression showed increased levels of P14780 gelatinase , motility in vitro , and metastatic potential in vivo . Expression of P14780 in the various cell clones was also negatively modulated by cell density , but this effect was much stronger in E-CD ( + ) cells , despite the fact that all of the cell clones analyzed maintained the expression of P22223 and made cell-cell contacts at high cell density . Our results indicate that in this cell system , the E-CD-mediated cell-cell contacts are involved in the down-regulation of P14780 expression . Thus , the loss of E-CD triggers a migratory and invasive phenotype in mouse squamous carcinoma cells . Hypoxia-induced neuroinflammatory white-matter injury reduced by minocycline in SHR/SP . Hypertensive small vessel disease is a major cause of vascular cognitive impairment ( VCI ) . Spontaneously hypertensive/stroke prone rats ( SHR/SP ) with unilateral carotid artery occlusion ( UCAO ) and a Japanese permissive diet ( JPD ) have white-matter ( WM ) damage similar to that seen in VCI . We hypothesized that WM injury was due to hypoxia-mediated , blood-brain barrier ( BBB ) disruption . Twelve-week-old SHR/SP had UCAO/JPD and were studied with immunohistochemistry , biochemistry , multimodal magnetic resonance imaging ( Q9BWK5 ) , and Morris water maze ( MWM ) testing . One week after UCAO/JPD , WM showed a significant increase in hypoxia inducible factor-1α ( HIF-1α ) , which increased further by 3 weeks . Prolyl hydroxylase-2 ( Q9GZT9 ) expression decreased at 1 and 3 weeks . Infiltrating T cells and neutrophils appeared around endothelial cells from 1 to 3 weeks after UCAO/JPD , and matrix metalloproteinase-9 ( P14780 ) colocalized with inflammatory cells . At 3 weeks , WM immunostained for IgG , indicating BBB leakage . DB01017 ( 50 mg/kg intraperitoneally ) was given every other day from weeks 12 to 20 . Multimodal Q9BWK5 showed that treatment with minocycline significantly reduced lesion size and improved cerebral blood flow . DB01017 improved performance in the MWM and prolonged survival . We propose that BBB disruption occurred secondary to hypoxia , which induced an P14780 -mediated infiltration of leukocytes . DB01017 significantly reduced WM damage , improved behavior , and prolonged life . DB01017 inhibits angiogenesis in vitro through the translational suppression of HIF-1α . DB01017 was recently found to be effective against cancer . However , the precise molecular mechanisms of minocycline in cancer are poorly understood . Hypoxia-inducible factor-1 ( Q9BYW2 , a heterodimeric transcription factor composed of HIF-1α and β ) activates the transcription of genes that are involved in angiogenesis in cancer . In this study , we found that minocycline significantly inhibits HIF-1α protein expression and suppresses Q9BYW2 transcriptional activity . The tube formation assay showed that minocycline has anti-angiogenic activity and suppresses hypoxia-induced vascular endothelial growth factor ( P15692 ) expression . The metabolic labeling assay showed that minocycline reduces HIF-1α protein translation and global protein synthesis . In addition , minocycline suppresses P42345 signaling and increases the phosphorylation of eIF2α , which is known to be related to the translational regulation of HIF-1α expression . These findings collectively indicate that minocycline is a potential inhibitor of HIF-1α and provide new insight into the discovery of drugs for cancer treatment . Gene set enrichment analysis unveils the mechanism for the phosphodiesterase 4B control of glucocorticoid response in B-cell lymphoma . PURPOSE : Resistance to glucocorticoid ( GC ) is a significant problem in the clinical management of lymphoid malignancies . Addressing this issue via a mechanistic understanding of relevant signaling pathways is more likely to yield positive outcomes . EXPERIMENTAL DESIGN : We used gene set enrichment analysis ( GSEA ) , multiple genetic models of gain and loss of function in B-cell lymphoma cell lines , in vitro and in vivo , and primary patient samples to characterize a novel relationship between the cyclic AMP/phosphodiesterase 4B ( DB02527 / Q07343 ) , AKT/ P42345 activities , and GC responses . RESULTS : Starting from the GSEA , we found that overexpression of the Q07343 in diffuse large B-cell lymphoma ( DLBCL ) impinge on the same genes/pathways that are abnormally active in GC-resistant tumors . We used genetically modified cell lines to show that Q07343 modulates DB02527 inhibitory activities toward the AKT/ P42345 pathway and defines GC resistance in DLBCL . In agreement with these data , pharmacologic inhibition of DB05876 in a xenograft model of human lymphoma unleashed DB02527 effects , inhibited AKT , and restored GC sensitivity . Finally , we used primary DLBCL samples to confirm the clinical relevance and biomarker potential of AKT/ P42345 regulation by Q07343 . CONCLUSIONS : Together , these data mechanistically elucidated how DB02527 modulates GC responses in lymphocytes , defined AKT as the principal transducer of the growth inhibitory effects of DB02527 in B cells , and allowed the formulation of genomics-guided clinical trials that test the ability of DB05876 inhibitors to restore GC sensitivity and improve the outcome of patients with B-cell malignancies . Selenium improves stem cell potency by stimulating the proliferation and active migration of 3T3- Q9NUQ9 preadipocytes . Selenium is a trace nutrient element that protects cells against oxidative damage . In this study , the potential of selenium to improve stem cell potency through active proliferation and migration of 3T3- Q9NUQ9 preadipocytes was investigated , together with the underlying molecular mechanisms . The results indicated that selenium applied for 24 h stimulated cell proliferation up to 20 % compared to untreated control cells . Selenium induced the expression of cyclin-dependent kinase ( CDK ) 1 and P24941 , which are known to regulate G2/M progression , and significantly downregulated the CDK inhibitors P38936 and p27 . Selenium also activated the expression of the phosphatidylinositol 3-kinase ( PI3K ) /Akt pathway , as well as extracellular signal-regulated kinase ( P29323 ) . Although LY294002 , an inhibitor of PI3K , significantly inhibited the selenium-induced cell proliferation of the 3T3- Q9NUQ9 preadipocytes , PD98059 , an inhibitor of P29323 , did not affect selenium-induced active proliferation . These results clearly indicate that selenium stimulated cell proliferation through cell cycle progression and PI3K/Akt activation , but not through P29323 activation . Furthermore , selenium increased 3T3- Q9NUQ9 cell migration , which was associated with the induction of matrix metalloproteinase ( MMP ) -2 and P14780 . Taken together , the current findings suggest that selenium can stimulate stem cell potency by increasing the proliferation and active migration of 3T3- Q9NUQ9 cells . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . Effects of zileuton and montelukast in mouse experimental spinal cord injury . BACKGROUND AND PURPOSE : P09917 ( P09917 ) is the key enzyme in leukotriene ( LT ) biosynthesis from arachidonic acid ( AA ) . Here , we examined the role of the P09917 -product , cysteinyl-LT ( DB00151 -LT ) , with a P09917 inhibitor ( zileuton ) and a DB00151 -LT , receptor antagonist ( montelukast ) , in the inflammatory response and tissue injury associated with spinal cord injury ( SCI ) . EXPERIMENTAL APPROACH : SCI was induced in mice by the application of vascular clips to the dura via a two-level Q8NHM4 to T7 laminectomy for 1 min . Cord inflammation was assessed histologically and by measuring inflammatory mediators ( ELISA ) and apoptosis by annexin V , TUNEL , P48023 staining and Bax and Bcl-2 expression ( immunohistochemistry and western blots ) . Motor function in hindlimbs was assessed by a locomotor rating scale , for 10 days after cord injury . KEY RESULTS : SCI in mice resulted in tissue damage , oedema , neutrophil infiltration , apoptosis , tumour necrosis-alpha ( P01375 ) and cyclooxygenase-2 ( P35354 ) expression , prostaglandin E(2) ( PGE(2) ) and leukotriene B(4) ( Q06643 (4) ) production , and extracellular signal-regulated kinase 1/2 ( P27361 /2 ) phosphorylation in injured tissue . Treatment of the mice with zileuton or montelukast reduced the spinal cord inflammation and tissue injury , neutrophil infiltration , P01375 , P35354 and pERK1/2 expression , PGE(2) and Q06643 (4) production , and apoptosis . In separate experiments , zileuton or montelukast significantly improved the recovery of limb function over 10 days . CONCLUSIONS AND IMPLICATIONS : Zileuton and montelukast produced a substantial reduction of inflammatory events associated with experimental SCI . Our data underline the important role of P09917 and DB00151 -LT in neurotrauma . Changes of Dietary Fat and Carbohydrate Content Alter Central and Peripheral Clock in Humans . CONTEXT : The circadian clock coordinates numerous metabolic processes with light-dark and feeding regimens . However , in humans it is unknown whether dietary patterns influence circadian rhythms . OBJECTIVE : We examined the effects of switching from a high-carbohydrate , low-fat diet to a low-carbohydrate , high fat ( LC/HFD ) isocaloric diet on the central and peripheral circadian clocks in humans . DESIGN : Diurnal patterns of salivary cortisol and gene expression were analyzed in blood monocytes of 29 nonobese healthy subjects before and 1 and 6 weeks after the dietary switch . For this , we established a method of rhythm prediction by 3-time point data . RESULTS : The centrally driven cortisol rhythm showed a phase delay 1 and 6 weeks after the dietary switch to a LC/HFD as well as an amplitude increase . The dietary switch altered diurnal oscillations of core clock genes ( O15534 , O15055 , P56645 , and Q10587 ) and inflammatory genes ( P08571 , Q99467 , P25963 , and P01584 ) . The LC/HFD also affected the expression of nonoscillating genes contributing to energy metabolism ( Q96EB6 ) and fat metabolism ( O15254 and P50213 ) . Expression of clock genes but not of salivary cortisol in monocytes tightly correlated with levels of blood lipids and with expression of metabolic and inflammatory genes . CONCLUSIONS : Our results suggest that the modulation of the dietary fat and carbohydrate content alters the function of the central and peripheral circadian clocks in humans . Molecular discrimination of responders and nonresponders to anti- P01375 alpha therapy in rheumatoid arthritis by etanercept . INTRODUCTION : About 30 % of rheumatoid arthritis patients fail to respond adequately to TNFalpha-blocking therapy . There is a medical and socioeconomic need to identify molecular markers for an early prediction of responders and nonresponders . METHODS : RNA was extracted from peripheral blood mononuclear cells of 19 rheumatoid arthritis patients before the first application of the TNFalpha blocker etanercept as well as after 72 hours . Clinical response was assessed over 3 months using the 28-joint-count Disease Activity Score and X-ray scans . Supervised learning methods were applied to Affymetrix Human Genome U133 microarray data analysis to determine highly selective discriminatory gene pairs or triplets with prognostic relevance for the clinical outcome evinced by a decline of the 28-joint-count Disease Activity Score by 1.2 . RESULTS : Early downregulation of expression levels secondary to TNFalpha neutralization was associated with good clinical responses , as shown by a decline in overall disease activity 3 months after the start of treatment . Informative gene sets include genes ( for example , P25963 , P13236 , P10145 , P01584 , P21580 , Q07343 , O75807 and P35318 ) involved in different pathways and cellular processes such as TNFalpha signalling via NFkappaB , NFkappaB-independent signalling via DB02527 , and the regulation of cellular and oxidative stress response . Pairs and triplets within these genes were found to have a high prognostic value , reflected by prediction accuracies of over 89 % for seven selected gene pairs and of 95 % for 10 specific gene triplets . CONCLUSION : Our data underline that early gene expression profiling is instrumental in identifying candidate biomarkers to predict therapeutic outcomes of anti-TNFalpha treatment regimes . Experimental models of neuroprotection relevant to multiple sclerosis . Activated T cells , particularly those of the T-helper ( Th ) 1 subset , have the capacity to kill neurons . Strategies for preventing such damage may include deviation of activated T cells into the Th2 subset ( e.g. , via use of glatiramer acetate ) , alteration of functional properties of Th1 cells ( e.g. , through use of interferon [ P27352 ] -beta or IV immunoglobulin ) , and inhibition of activated cell migration into the CNS ( e.g. , by employing P27352 -beta or natalizumab ) . P14780 ( P14780 ) also causes neuron death in neurotoxicity models , and examination of medications with MMP inhibitory activity indicates that minocycline is capable of preventing such damage . DB01017 also has other properties relevant to conferring neuroprotection , such as inhibition of microglial activity and apoptosis pathways . In a small pilot study in patients with relapsing-remitting multiple sclerosis , minocycline treatment produced favorable outcomes in terms of gadolinium-enhancing lesions and clinical course . Further studies are needed to establish whether experimental neuroprotection strategies involving these mechanisms may be translated into preventing neurodegeneration in multiple sclerosis . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Pharmacogenetics of asthma . PURPOSE OF REVIEW : Patient response to the asthma drug classes , bronchodilators , inhaled corticosteroids and leukotriene modifiers , are characterized by a large degree of heterogeneity , which is attributable in part to genetic variation . Herein , we review and update the pharmacogenetics and pharmaogenomics of common asthma drugs . RECENT FINDINGS : Early studies suggest that bronchodilator reversibility and asthma worsening in patients on continuous short-acting and long-acting beta-agonists are related to the Gly16Arg genotype for the P07550 . More recent studies including genome-wide association studies implicate variants in other genes contribute to bronchodilator response heterogeneity and fail to replicate asthma worsening associated with continuous beta-agonist use . Genetic determinants of the safety of long-acting beta-agonist require further study . Variants in P34998 , Q9UL17 , and P06734 contribute to variability in response for lung function , airways responsiveness , and exacerbations in patients taking inhaled corticosteroids . Variants in P09917 , P09960 , Q16873 , P33527 , Q9NS75 , and O94956 contribute to variability in response to leukotriene modifiers . SUMMARY : Identification of novel variants that contribute to response heterogeneity supports future studies of single nucleotide polymorphism discovery and include gene expression and genome-wide association studies . Statistical models that predict the genomics of response to asthma drugs will complement single nucleotide polymorphism discovery in moving toward personalized medicine . Q9GZV9 and matrix-metalloproteinases in patients with chronic kidney disease : are they associated with cardiovascular disease ? BACKGROUND : High cardiovascular risk in patients with chronic kidney disease ( CKD ) may be related to mineral disorder and microinflammation . Q9GZV9 ( Q9GZV9 ) is a phosphatonin and inhibitor of calcitriol synthesis , which is associated with poor prognosis in CKD patients starting dialysis . Matrix-metalloproteinases ( P08253 , P14780 ) contribute to myocardial remodeling and arterial calcification . Q9GZV9 and MMPs levels are altered in CKD , however , little is known about their association and relation to cardiovascular ( CV ) disease . METHODS : Standard laboratory parameters , plasma levels of P08253 , P14780 , Q9GZV9 , Q13219 and CV disease history were assessed in 80 patients with CKD 1-5 and 44 healthy control subjects . RESULTS : Q9GZV9 and P08253 ( assessed by ELISA ) were higher in CKD patients compared to controls . Q9GZV9 increased from CKD 3 , whereas P08253 increased only in CKD 5 . Q9GZV9 was positively associated with P08253 , adjusted to age , eGFR , phosphatemia , calcitriol and parathormone . Q9GZV9 independently correlated with parathormone and inversely with calcitriol , whereas P08253 was related to phosphatemia . Q9GZV9 was higher in subjects with a history of CV disease compared to those free of such history ( 559.0 vs.184.0 RU/ml ) , adjusted to age and eGFR . CONCLUSION : Our data suggest a possible relationship between Q9GZV9 , P08253 and CV disease in CKD . Potential causality of this association remains to be elucidated . Association of common genetic variants with risperidone adverse events in a Spanish schizophrenic population . DB00734 non-compliance is often high due to undesirable side effects , whose development is in part genetically determined . Studies with genetic variants involved in the pharmacokinetics and pharmacodynamics of risperidone have yielded inconsistent results . Thus , the aim of this study was to investigate the putative association of genetic markers with the occurrence of four frequently observed adverse events secondary to risperidone treatment : sleepiness , weight gain , extrapyramidal symptoms and sexual adverse events . A series of 111 schizophrenia inpatients were genotyped for genetic variants previously associated with or potentially involved in risperidone response . Presence of adverse events was the main variable and potential confounding factors were considered . Allele 16Gly of P07550 was significantly associated with a higher risk of sexual adverse events . There were other non-significant trends for P35462 9Gly and P31645 S alleles . Our results , although preliminary , provide new candidate variants of potential use in risperidone safety prediction . DB01017 attenuates hypoxia-inducible factor-1α expression correlated with modulation of p53 and AKT/ P42345 /p70S6K/ Q13541 pathway in ovarian cancer : in vitro and in vivo studies . Hypoxia-inducible factor ( HIF ) -1α is the key cellular survival protein under hypoxia , and is associated with tumor progression and angiogenesis . We have recently shown the inhibitory effects of minocycline on ovarian tumor growth correlated with attenuation of vascular endothelial growth factor ( P15692 ) and herein report a companion laboratory study to test if these effects were the result of HIF-1α inhibition . In vitro , human ovarian carcinoma cell lines ( A2780 , OVCAR-3 and SKOV-3 ) were utilized to examine the effect of minocycline on Q9BYW2 and its upstream pathway components to elucidate the underlying mechanism of action of minocycline . Mice harboring OVCAR-3 xenografts were treated with minocycline to assess the in vivo efficacy of minocycline in the context of Q9BYW2 . DB01017 negatively regulated HIF-1α protein levels in a concentration-dependent manner and induced its degradation by a mechanism that is independent of prolyl-hydroxylation . The inhibition of HIF-1α was found to be associated with up-regulation of endogenous p53 , a tumor suppressor with confirmed role in HIF-1α degradation . Further studies demonstrated that the effect of minocycline was not restricted to proteasomal degradation and that it also caused down-regulation of HIF-1α translation by suppressing the AKT/ P42345 /p70S6K/ Q13541 signaling pathway . DB01017 treatment of mice bearing established ovarian tumors , led to suppression of HIF-1α accompanied by up-regulation of p53 protein levels and inactivation of AKT/ P42345 /p70S6K/ Q13541 pathway . These data reveal the therapeutic potential of minocycline in ovarian cancer as an agent that targets the pro-oncogenic factor HIF-1α through multiple mechanisms . DB01017 increases phosphorylation and membrane insertion of neuronal GluR1 receptors . The tetracycline antibiotic minocycline beneficially affects neuronal functioning and also inhibits the enzyme P09917 ( 5- P28300 ) . We hypothesized that similar to 5- P28300 inhibitors , minocycline may increase phosphorylation and membrane insertion of the glutamate receptor GluR1 . The experiments were performed in primary cultures of mouse striatal neurons and in the prefrontal cortex and striatum of minocycline-treated mice . In vitro , low micromolar minocycline concentrations increased GluR1 phosphorylation at Ser845 and Ser831 and increased the surface content of GluR1 . DB01017 also increased GluR1 phosphorylation in vivo . Increased GluR1 phosphorylation and minocycline treatment have been associated with antidepressant and memory-enhancing activities . Direct consequences of minocycline-increased GluR1 phosphorylation are yet to be established . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . Type I saikosaponins a and d inhibit osteoclastogenesis in bone marrow-derived macrophages and osteolytic activity of metastatic breast cancer cells . Many osteopenic disorders , including a postmenopausal osteoporosis and lytic bone metastasis in breast and prostate cancers , are linked with a hyperosteoclast activity due to increased receptor activator of nuclear factor kappa-B ligand ( O14788 ) expression in osteoblastic/stromal cells . Therefore , inhibition of O14788 -induced osteoclastogenesis and osteoclast-induced bone resorption is an important approach in controlling pathophysiology of these skeletal diseases . We found that , of seven type I , II , and III saikosaponins isolated from Bupleurum falcatum , saikosaponins A and D , type I saikosaponins with an allyl oxide linkage between position 13 and 28 and two carbohydrate chains that are directly attached to the hydroxyl groups in position 3 , exhibited the most potent inhibition on O14788 -induced osteoclast formation at noncytotoxic concentrations . The stereochemistry of the hydroxyl group at C16 did not affect their activity . Saikosaponins A and D inhibited the formation of resorptive pits by reducing the secreted levels of matrix metalloproteinase- ( MMP- ) 2 , P14780 , and cathepsin K in O14788 -induced osteoclasts . Additionally , saikosaponins A and D inhibited mRNA expression of parathyroid hormone-related protein as well as cell viability and invasion in metastatic human breast cancer cells . Thus , saikosaponins A and D can serve as a beneficial agent for the prevention and treatment of osteoporosis and cancer-induced bone loss . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Protection of minocycline on early brain injury after subarachnoid hemorrhage in rats . DB01017 has been shown to be neuroprotective in cerebral ischemia and in other models of brain injury . Our goal is to observe the protection of minocycline on EBI after Q53FZ2 and the mechanism . 48 adult male SD rats were randomly divided into four groups : the sham-operated group , Q53FZ2 group , vehicle group ( Q53FZ2 +normal sodium ) , and minocycline group ( Q53FZ2 +minocycline ) . The Q53FZ2 model was induced by injecting 300 μl of autologous arterial blood into the prechiasmatic cistern . Expressions of P14780 in the hippocampus were examined at 24 h by western blot and zymography . Western blot and zymography showed that the expression of total and active P14780 increased dramatically at 24 h after Q53FZ2 compared with that of the sham group ( P < 0.01 ) . The clinical assessments got a lower score than that of the sham-operated group . After treated with minocycline , the expression of P14780 decreased significantly ( P < 0.01 vs. vehicle group ) , and the clinical assessments improved . We conclude that minocycline can protect EBI after Q53FZ2 , which may be related to the mechanism of inhibiting the expression of P14780 in the hippocampus . Involvement of matrix metalloproteinases-2 and -9 in the formation of a lacuna-like cerebral cavity . We used a modified pial vessel disruption ( PVD ) protocol with adult male Wistar rats to mimic small-vessel stroke in the cerebral cortex . Within 3 weeks , this lesion develops into a single lacuna-like cavity , which is fluid-filled and encapsulated by reactive astrocytes . DB01017 treatment that commences 1 hr after lesion and continues for 6 days prevents the cavitation and causes a filling of the lesion with reactive astrocytes and no barrier . Here , we determined whether inhibition of matrix metalloproteinases-2 and -9 ( MMPs ) mediates this minocycline action . Confocal microscopy revealed increased punctate staining of MMPs inside the lesion sites after 2 days of PVD . Astrocytes lined the lesion border but showed sparse localization inside the lesion . In contrast , increased MMP levels inside the lesion coincided with increased Q92838 or OX-42 immunostaining , suggesting that MMP elevation reflected increased secretions from microglia/macrophages . Imaging analyses also revealed that minocycline administered for 2 days before animal euthanasia , significantly decreased MMP levels within the lesion . Moreover , Western blot analysis of cortical tissue extracts showed a significant 30-40 % upregulation of MMPs 2 days after lesion . DB01017 administered 2 hr before the lesion significantly inhibited both P14780 and P08253 levels by ∼40 % . In contrast , minocycline administered 1 hr after the lesion only decreased P14780 levels by ∼30 % . Because MMP inhibition with batimastat injection also prevented cavity formation at 21 days , we conclude that minocycline prevented the creation of a lacuna-like cyst in the cerebral cortex by inhibiting the MMP secretion from microglia in the affected tissue . P10275 promotes esophageal cancer cell migration and proliferation via matrix metalloproteinase 2 . Esophageal squamous cell carcinoma ( ESCC ) is one of the most common malignancies worldwide . P10275 ( AR ) plays an important role in many kinds of cancers . However , the molecular mechanisms of AR in ESCC are poorly characterized . In the present study , Western blot analysis and real-time quantitative PCR were performed to identify differentially expressed AR in 40 ESCC tissue samples , which revealed that the messenger RNA ( mRNA ) and protein expression of AR is upregulated in the ESCC tissue samples . AR overexpression induced increases in ESCC cell invasion and proliferation in vitro . Silencing of AR inhibited the proliferation of KYSE450 cells which have a relatively high level of AR , and the invasion of KYSE450 cells was distinctly suppressed . Furthermore , AR knockdown led to substantial reductions in matrix metalloproteinase 2 ( P08253 ) and p-AKT levels in ESCC cell lines , but no significant change in AKT and P14780 expression . These results suggest that AR is involved in tumor progression , and thus , AR could represent selective targets for the molecularly targeted treatments of ESCC . DB01017 with aspirin : a therapeutic approach in the treatment of diabetic neuropathy . Enhanced production of matrix metalloproteinase-2 ( P08253 ) and matrix metalloproteinase-9 ( P14780 ) in diabetes leads to degradation of extracellular matrix in blood vessels and leads to complications of diabetes . In the present study , we have targeted P08253 and P14780 overactivation in diabetic neuropathy using a known P08253 and P14780 inhibitor , minocycline , with a non-selective P36551 inhibitor , aspirin . DB00428 -induced diabetic neuropathy was carried out in male Wistar rats and monitored by measuring the sensory nerve conduction velocity ( SNCV ) , motor nerve conduction velocity ( MNCV ) , tail flick latency and hot plate latency . Three weeks of treatment with a combination of minocycline and aspirin showed significant improvement in SNCV , MNCV , hot plate latency and tail flick latency when compared with diabetic control . The results of the present study suggest that P08253 and P14780 inhibition in the presence of P36551 inhibitor prevents the development of experimental diabetic neuropathy in rats and can be a potential approach for the treatment . P06850 family of peptides regulates intestinal angiogenesis . BACKGROUND & AIMS : The corticotrophin-releasing hormone ( P06850 ) family of peptides modulates intestinal inflammation and the P06850 receptor 2 ( Q13324 ) suppresses postnatal angiogenesis in mice . We investigated the functions of P34998 and Q13324 signaling during intestinal inflammation and angiogenesis . METHODS : The activities of P34998 and Q13324 were disrupted by genetic deletion in mice or with selective antagonists . A combination of in vivo , ex vivo , and in vitro measures of angiogenesis were used to determine their activity . P34998 (-/-) mice and Q13324 (-/-) mice with dextran sodium sulfate-induced colitis were analyzed in comparison with wild-type littermates ( controls ) . RESULTS : Colitis was significantly reduced in mice in which P34998 activity was disrupted by genetic deletion or with an antagonist , determined by analyses of survival rate , weight loss , histological scores , and cytokine production . Inflammation was exacerbated in mice in which Q13324 activity was inhibited by genetic deletion or with an antagonist , compared with controls . The inflamed intestines of P34998 (-/-) mice had reduced microvascular density and expression of vascular endothelial growth factor ( P15692 ) -A , whereas the intestines of Q13324 (-/-) mice had increased angiogenesis and P15692 levels . An antagonist of P35968 activity alleviated colitis in Q13324 (-/-) mice . Ex vivo aortic vessel outgrowth was reduced when P34998 was deficient but increased when Q13324 was deficient . The P34998 preferred agonist P06850 stimulated tube formation , proliferation , and migration of cultured intestinal microvascular endothelial cells by phosphorylating Akt , whereas the specific Q13324 agonist Q969E3 had opposite effects . CONCLUSION : P34998 promotes intestinal inflammation , as well as endogenous and inflammatory angiogenesis whereas Q13324 inhibits these activities . PEG minocycline-liposomes ameliorate CNS autoimmune disease . BACKGROUND : DB01017 is an oral tetracycline derivative with good bioavailability in the central nervous system ( CNS ) . DB01017 , a potent inhibitor of matrix metalloproteinase ( MMP ) -9 , attenuates disease activity in experimental autoimmune encephalomyelitis ( EAE ) , an animal model of multiple sclerosis ( MS ) . Potential adverse effects associated with long-term daily minocycline therapy in human patients are concerning . Here , we investigated whether less frequent treatment with long-circulating polyethylene glycol ( PEG ) minocycline liposomes are effective in treating EAE . FINDINGS : Performing in vitro time kinetic studies of PEG minocycline-liposomes in human peripheral blood mononuclear cells ( PBMCs ) , we determined that PEG minocycline-liposome preparations stabilized with CaCl(2) are effective in diminishing P14780 activity . Intravenous injections of PEG minocycline-liposomes every five days were as effective in ameliorating clinical EAE as daily intraperitoneal injections of minocycline . Treatment of animals with PEG minocycline-liposomes significantly reduced the number of CNS-infiltrating leukocytes , and the overall expression of P14780 in the CNS . There was also a significant suppression of P14780 expression and proteolytic activity in splenocytes of treated animals , but not in CNS-infiltrating leukocytes . Thus , leukocytes gaining access to the brain and spinal cord require the same absolute amount of P14780 in all treatment groups , but minocycline decreases the absolute cell number . CONCLUSIONS : Our data indicate that less frequent injections of PEG minocycline-liposomes are an effective alternative pharmacotherapy to daily minocycline injections for the treatment of CNS autoimmune diseases . Also , inhibition of P14780 remains a promising treatment target in EAE and patients with MS . DB01017 prevents dynorphin-induced neurotoxicity during neuropathic pain in rats . Despite many advances , our understanding of the involvement of prodynorphin systems in the development of neuropathic pain is not fully understood . Recent studies suggest an important role of neuro-glial interactions in the dynorphin effects associated with neuropathic pain conditions . Our studies show that minocycline reduced prodynorphin mRNA levels that were previously elevated in the spinal and/or dorsal root ganglia ( Q86YR7 ) following sciatic nerve injury . The repeated intrathecal administration of minocycline enhanced the analgesic effects of low-dose dynorphin ( 0.15 nmol ) and U50,488H ( 25-100 nmol ) and prevented the development of flaccid paralysis following high-dose dynorphin administration ( 15 nmol ) , suggesting a neuroprotective effect . DB01017 reverts the expression of IL-1β and P05231 mRNA within the spinal cord and IL-1β mRNA in Q86YR7 , which was elevated following intrathecal administration of dynorphin ( 15 nmol ) . These results suggest an important role of these proinflammatory cytokines in the development of the neurotoxic effects of dynorphin . Similar to minocycline , a selective inhibitor of P14780 ( P14780 levels are reduced by minocycline ) exerts an analgesic effect in behavioral studies , and its administration prevents the occurrence of flaccid paralysis caused by high-dose dynorphin administration ( 15 nmol ) . In conclusion , our results underline the importance of neuro-glial interactions as evidenced by the involvement of IL-1β and P05231 and the minocycline effect in dynorphin-induced toxicity , which suggests that drugs that alter the prodynorphin system could be used to better control neuropathic pain . Autosomal-dominant hypophosphatemic rickets ( P30518 ) mutations stabilize Q9GZV9 . BACKGROUND : The gene for the renal phosphate wasting disorder autosomal-dominant hypophosphatemic rickets ( P30518 ) is Q9GZV9 , which encodes a secreted protein related to the fibroblast growth factors ( FGFs ) . We previously detected missense mutations R176Q , R179W , and R179Q in Q9GZV9 from P30518 kindreds . The mutations replace R residues within a subtilisin-like proprotein convertase ( Q969E3 ) cleavage site 176RHTR-179 ( RXXR motif ) . The goal of these studies was to determine if the P30518 mutations lead to protease resistance of Q9GZV9 . METHODS : The P30518 mutations were introduced into human Q9GZV9 cDNA clones with or without an N-terminal FLAG tag by site-directed mutagenesis and were transiently transfected into HEK293 cells . Protein expression was determined by Western analyses . RESULTS : Antibodies directed toward the C-terminal portion of Q9GZV9 revealed that the native Q9GZV9 protein resolved as 32 kD and 12 kD species in HEK293 conditioned media ; however , the three mutated proteins were detected only as the 32 kD band . An N-terminal FLAG-tagged native Q9GZV9 resolved as two bands of 36 kD and 26 kD when detected with a FLAG antibody , whereas the R176Q mutant resolved primarily as the 36 kD protein species . Cleavage of Q9GZV9 was not enhanced by extracellular incubation of Q9GZV9 with HEK293 cells . Native and mutant FGF-23s bound heparin . CONCLUSIONS : Q9GZV9 proteins containing the P30518 mutations are secreted , and produce polypeptides less sensitive to protease cleavage than wild-type Q9GZV9 . Therefore , the P30518 mutations may protect Q9GZV9 from proteolysis , thereby potentially elevating circulating concentrations of Q9GZV9 and leading to phosphate wasting in P30518 patients . Transient estrogen exposure from birth affects uterine expression of developmental markers in neonatal gilts with lasting consequences in pregnant adults . Disruption of estrogen-sensitive , estrogen receptor ( ER ) -dependent events during porcine uterine development between birth ( postnatal day= P01160 0 ) and P01160 14 affects patterns of uterine morphoregulatory gene expression in the neonate with lasting consequences for reproductive success . Uterine capacity for conceptus support is reduced in pregnant adult gilts exposed to estradiol valerate ( EV ) for 14 days from birth . Objectives here were to determine effects of EV exposure from birth through P01160 13 on neonatal uterine and adult endometrial markers of growth , patterning , and remodeling . Targets included the relaxin receptor ( Q9HBX9 ) , estrogen receptor-alpha ( P03372 ) and vascular endothelial growth factor ( P15692 ) , morphoregulatory markers P31260 and O00755 , and the matrix metalloproteinases (MMP)2 and P14780 . Gilts were treated daily with EV ( 50 microg/kg body weight per day , i.m. ) or corn oil vehicle from birth through P01160 13 . Uteri were obtained from neonates on P01160 14 and from adults on pregnancy day 12 ( PxD 12 ) . In neonates , EV exposure from birth increased uterine Q9HBX9 gene expression , and both P03372 and P15692 proteins . At PxD 12 , endometrial Q9HBX9 mRNA remained elevated , while P03372 protein was reduced . Early EV treatment decreased neonatal uterine O00755 , but increased P31260 expression . O00755 expression was reduced in EV-treated adults . Transient EV exposure increased P14780 transcripts at P01160 14 , whereas both latent and active P14780 activity was increased due to early EV treatment in adults on PxD 12 . Results support the hypothesis that transient , estrogen-induced disruption of porcine uterine development from birth alters early programming events that lead to functional consequences in the adult . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . DB01017 reduces renal microvascular leakage in a rat model of ischemic renal injury . Tetracyclines exhibit significant anti-inflammatory properties , inhibit matrix metalloproteinases ( MMPs ) , and are protective in models of ischemia-reperfusion injury ( IRI ) . Both inflammatory cascades and MMP activation have been demonstrated to modulate microvascular permeability . Because increased microvascular permeability occurs during IRI in a variety of organ systems including the kidney , we hypothesized that minocycline , a semisynthetic tetracycline , would diminish microvascular leakage during renal IRI . To test this hypothesis , we used intravital 2-photon microscopy to examine leakage of fluorescent dextrans from the vasculature in a rodent model of IRI . DB01017 significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Although minocycline diminished leukocyte accumulation in the kidney following ischemia , areas of leukocyte accumulation did not correlate with areas of microvascular permeability in either the saline- or minocycline-pretreated animals . DB01017 diminished the perivascular increase in P08253 and P14780 , as well as the increase in P08253 activity 24 h after ischemia . ABT-518 , a specific inhibitor of P08253 and P14780 , also significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Our results indicate that minocycline mitigates the renal microvascular permeability defect following IRI . This effect is spatially distinct from the effect of minocycline on leukocyte accumulation and may be related to diminished activity of MMPs on the integrity of the perivascular matrix . DB01017 protects against permanent cerebral ischemia in wild type but not in matrix metalloprotease-9-deficient mice . DB01017 is protective in models of transient middle cerebral artery occlusion ( MCAO ) . We studied whether minocycline and doxycycline , another tetracycline derivative , provide protection in permanent MCAO . Because minocycline inhibits matrix metalloprotease-9 ( P14780 ) , we also compared minocycline 's protective effect in wild type ( wt ) and P14780 knock-out ( ko ) mice . Wt FVB/N , Balb/C , and two lines of P14780 ko and their wt C57Bl/6 control mice were subjected to 24- or 72-hour permanent MCAO . Drug administration was started either 12 hours before or 2 hours after the onset of MCAO . Infarct size was determined by triphenyltetrazolium staining or P24752 -weighted Q9BWK5 . Zymography was used to study the expression of MMPs . In wt strains , tetracycline treatments started before MCAO reduced the infarct size by 25 % to 50 % , whereas the treatment started after MCAO was not protective . DB01017 inhibited ischemia-provoked pro- P14780 induction in wt mice , but was not protective in P14780 ko mice . Pro- P08253 was induced by MCAO in wt and P14780 ko mice . MCAO-induced pro- P08253 was downregulated by minocycline treatment in wt mice but remained in P14780 ko mice at the same level as in saline-treated wt mice . Tetracyclines are protective in permanent MCAO when the treatment is started before the insult . DB01017 may provide protection by interfering with MMPs . Clinical and pathogenic aspects of candidate genes for lithium prophylactic efficacy . A number of candidate genes for lithium prophylactic efficacy have been proposed , some of them being also associated with a predisposition to bipolar illness . The aim of the present study was to investigate a possible association between polymorphisms of 14 common genes with the quality of prophylactic lithium response in patients with bipolar mood disorder , in relation to the putative role of these genes in the pathogenesis of this disorder . Some association with lithium prophylactic efficacy was found for the polymorphisms of P31645 , P21728 , P21964 , P23560 and P06241 genes , but not for 5HT2A , 5HT2C , P14416 , P35462 , P21917 , GSK-3 , Q16620 , Q13224 and P14780 . Possible aspects of these genes with regard to the mechanism of lithium activity and pathogenesis of bipolar mood disorder are discussed . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . DB01017 suppresses experimental autoimmune encephalomyelitis by increasing tissue inhibitors of metalloproteinases . Matrix metalloproteinases ( MMPs ) that are secreted by activated T cells play a significant role in degradation of the extracellular matrix around the blood vessels and facilitate autoimmune neuroinflammation ; however , it remains unclear how MMPs act in lesion formation and whether MMP-targeted therapies are effective in disease suppression . In the present study , we attempted to treat experimental autoimmune encephalomyelitis ( EAE ) by administration of small interfering RNAs ( siRNAs ) for P08253 , P14780 , and minocycline , all of which have MMP-inhibiting functions . DB01017 , but not siRNAs , significantly suppressed disease development . In situ zymography revealed that gelatinase activities were almost completely suppressed in the spinal cords of minocycline-treated animals , while significant gelatinase activities were measured in the EAE lesions of control animals . However , P08253 and P14780 mRNAs and proteins in the spinal cords of treated rats were unexpectedly upregulated . At the same time , mRNA for tissue inhibitors of MMPs ( P01033 ) -1 and -2 were also upregulated . The EnzChek Gelatinase/Collagenase assay using tissue containing native MMPs and TIMPs demonstrated that gelatinase activity levels in the spinal cords of treated rats were suppressed to the same level as those in normal spinal cord tissues . Finally , double immunofluorescent staining demonstrated that P14780 immunoreactivities of treated rats were almost the same as those of control rats and that P14780 and P01033 immunoreactivities were colocalized in the spinal cord . These findings suggest that minocycline administration does not suppress MMPs at mRNA and protein levels but that it suppresses gelatinase activities by upregulating TIMPs . Thus , MMP-targeted therapies should be designed after the mechanisms of candidate drugs have been considered . Protective effect of beraprost sodium , a stable prostacyclin analog , in the development of cigarette smoke extract-induced emphysema . Chronic inflammation , imbalance of proteolytic and anti-proteolytic activities , oxidative stress , and apoptosis of lung structural cells contribute to the pathogenesis of P48444 . DB01240 protects cells against apoptosis , has anti-inflammatory properties , partially prevents cigarette smoke extract ( CSE ) -induced apoptosis of the pulmonary endothelium , and thus may be relevant in the pathogenesis of emphysema . We determined whether a synthetic stable prostacyclin analog , beraprost sodium ( BPS ) , attenuates the development of CSE-induced emphysema and elucidated the molecular mechanisms involved in its effect . Sprague-Dawley rats were treated with BPS and injected with CSE once a week for 3 wk . We measured the DNA damage of cells , the expression of caspase-3 , and the activity of matrix metalloproteinase ( MMP ) -2 and P14780 . We also analyzed TNFalpha and IL-1beta concentrations and the serum antioxidant activity . BPS prevented the development of CSE-induced emphysema , resulting in significant attenuation in alveolar enlargement and pulmonary parenchymal destruction . BPS inhibited pulmonary apoptosis and induction of P08253 and P14780 activity . Moreover , the protective effect of BPS was associated with a reduction of the expression of proinflammatory cytokines including TNFalpha and IL-1beta and a normalized biological oxidant activity . BPS introduces all these events , probably by activating DB02527 signaling through acting specific prostacyclin receptors . In conclusion , BPS protects against the development of CSE-induced emphysema by attenuating apoptosis , inhibiting proteolytic enzyme activity , reducing inflammatory cytokine levels , and augmenting antioxidant activity . BPS may potentially represent a new therapeutic option in the prevention of emphysema in humans in prospect . Rictor regulates P14780 activity and invasion through P04049 -MEK- P29323 signaling pathway in glioma cells . Glioblastoma multiforme ( GBM ) is the most common and highly aggressive type of primary brain tumor . Tumor-associated macrophages ( TAMs ) secrete P01375 -α that activates important survival pathways including Akt ( P31749 ) / P42345 network . The mammalian target of rapamycin ( P42345 ) network functions downstream of PI3K/Akt pathway to regulate cell growth , proliferation and survival . P42345 exists in two distinct complexes-mTORC1 and mTORC2 that differ in their components and sensitivity to rapamycin . The rapamycin-insensitive complex ( mTORC2 ) consists of P42345 , Q9BVC4 , Rictor , mSin1 and Protor and regulates the actin cytoskeleton in addition to activating Akt ( protein kinase B ) . The present study aimed to investigate the role of Rictor-a core component of mTORC2 in regulating proliferation , survival , and invasion in gliomas . siRNA-mediated loss of Rictor function in human glioma cell lines , LN18 and LN229 and in primary GBM cells resulted in elevated expression and activity of P14780 and significant increase in the invasive potential of these cells . Mechanistic studies revealed that the activation of P04049 -MEK- P29323 pathway was essential for induction of P14780 activity and enhanced invasion . Interestingly , ablation of Rictor did not affect P01375 -α-induced P14780 activity and invasiveness suggesting that P01375 -α in the microenvironment of tumor might overrule the function of Rictor as a negative regulator of P14780 and invasion . Silencing Rictor had no effect on the survival or proliferation in the cell lines in the presence or absence of P01375 -α . Our findings identify a role for Rictor in bridging two major pathways-Akt ( P31749 ) / P42345 and P04049 -MEK- P29323 in regulating P14780 activity and invasion of glioma tumor cells . P14416 occupancy by risperidone : implications for the timing and magnitude of clinical response . The objective of the study is to investigate whether dopamine D2 receptor occupancy by risperidone and plasma levels over time can account for therapeutic efficacy and the latency period to response . Thirty-eight examinations with (123)I-IBZM single photon emission computed tomography were performed on 22 patients with schizophrenia , at diagnosis , 48 h after starting risperidone treatment and at a stable dose . DB00734 plasma levels were determined and psychopathologic evaluations ( Brief Psychiatric Rating Scale , Positive and Negative Syndrome Scale ) were carried out . No differences in the striatal/occipital ( S/O ) ratio or plasma levels were found between examinations at the 48-h time point and when a stable dose level had been established , so these parameters could not account for the latency period required for clinical response . D2 receptor occupancy at 48 h correlated positively with clinical improvement after 2 weeks of treatment . Therefore , if these results are confirmed , D2 receptor occupancy at the beginning of treatment with risperidone may be a predictor of subsequent clinical response . Delivering minocycline into brain endothelial cells with liposome-based technology . DB01017 has been proposed as a way to blunt neurovascular injury from matrix metalloproteinases ( MMPs ) during stroke . However , recent clinical trials suggest that high levels of minocycline may have deleterious side-effects . Here , we showed that very high minocycline concentrations damage endothelial cells via calpain/caspase pathways . To alleviate this potential cytotoxicity , we encapsulated minocycline in liposomes . Low concentrations of minocycline could not reduce tumor necrosis factor α ( TNFα ) -induced P14780 release from endothelial cells . But low concentrations of minocycline-loaded liposomes significantly reduced TNFα-induced P14780 release . This study provides proof-of-concept that liposomes may be used to deliver lower levels of minocycline for targeting MMPs in cerebral endothelium . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood-lymphocytes . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood lymphocytes was investigated in vitro . The effects of the antibiotics on the mitotic response of purified lymphocytes stimulated with P01584 varied according to the individual from whom the lymphocytes were obtained . At concentrations above those reported to be present in serum during conventional therapy ( 2-8 mg/l ) , there was a tendency for both minocycline and tetracycline to suppress the mitotic response . DB01017 was superior to tetracycline in this respect . However , at physiological concentrations the antibiotics either had no significant effect , suppressed the mitotic response ( minocycline at 2 mg/l with one of six donors ) , or enhanced the mitotic response ( tetracycline at 2 and 8 mg/l with four of six donors ) . The stimulatory effect of tetracycline was not demonstrated when lymphocytes were cultured in whole blood for up to seven days with the antibiotic alone . Similar effects of the antibiotics were observed when mononuclear cell fractions isolated from six donors were stimulated with an optimal concentration of phytohaemagglutinin ( PHA ) . Stimulation of lymphocytes in whole blood cultures with PHA in the presence of minocycline and tetracycline revealed that , under these culture conditions , the antibiotics could suppress the mitotic response of lymphocytes at physiological doses with cells from a majority of donors . DB01017 improves functional outcomes , memory deficits , and histopathology after endovascular perforation-induced subarachnoid hemorrhage in rats . Subarachnoid hemorrhage ( Q53FZ2 ) results in significant long-lasting cognitive dysfunction . Therefore , evaluating acute and long-term outcomes after therapeutic intervention is important for clinical translation . The aim of this study was to use minocycline , a known neuroprotectant agent , to evaluate the long-term benefits in terms of neurobehavior and neuropathology after experimental Q53FZ2 in rats , and to determine which neurobehavioral test would be effective for long-term evaluation . Q53FZ2 was induced by endovascular perforation in adult male Sprague-Dawley rats ( n=118 ) . The animals were treated with intraperitoneal injection of minocycline ( 45 mg/kg or 135 mg/kg ) or vehicle 1 h after Q53FZ2 induction . In the short-term , animals were euthanized at 24 and 72 h for evaluation of neurobehavior , brain water content , and matrix metalloproteinase ( MMP ) activity . In the long-term , neurobehavior was evaluated at days 21-28 post- Q53FZ2 , and histopathological analysis was done at day 28 . High-dose but not low-dose minocycline reduced brain water content at 24 h , and therefore only the high-dose regimen was used for further evaluation , which reduced P14780 activity at 24 h . Further , high-dose minocycline improved spatial memory and attenuated neuronal loss in the hippocampus and cortex . The rotarod , T-maze , and water maze tests , but not the inclined plane test , detected neurobehavioral deficits in Q53FZ2 rats at days 21-28 . This study demonstrates that minocycline attenuates long-term functional and morphological outcomes after endovascular perforation-induced Q53FZ2 . Long-term neurobehavioral assessments using the rotarod , T-maze , and water maze tests could be useful to evaluate the efficacy of therapeutic intervention after experimental Q53FZ2 . DB01017 with aspirin : an approach to attenuate diabetic nephropathy in rats . Degradation of extracellular matrix ( Q13201 ) by enhanced production of matrix metalloproteinase-2 ( P08253 ) and matrix metalloproteinase-9 ( P14780 ) in diabetes leads to nephropathy . Cyclooxygenases ( P36551 ) further increase levels of these MMPs . The objective of present study was to inhibit P08253 and P14780 by combination of minocycline and aspirin to treat diabetic nephropathy . Diabetes was induced in male Wistar rats by streptozotocin ( Q11206 , 55 mg/kg i.p. ) . Four weeks after diabetes induction , the rats were treated with minocycline ( 50 mg/kg , p.o. ) , aspirin ( 50 mg/kg , p.o. ) , or minocycline ( 50 mg/kg , p.o. ) plus aspirin ( 50 mg/kg , p.o. ) for a period of 4 weeks . At the end of eighth week fluid input , urine output , and renal function tests were carried out for diagnosis of diabetic nephropathy . Renal hypertrophy was measured and histopathology was done to evaluate renal damage . Diabetes produced significant loss of body weight , polyuria , polydipsia , hyperglycemia , and increase in blood pressure . Serum creatinine , urea , and blood urea nitrogen levels were found to be increased significantly in the Q11206 group diabetic rats . Treatment with combination of minocycline and aspirin significantly prevented the rise in creatinine , urea , and blood urea nitrogen levels and increased creatinine clearance . Image analysis of kidneys revealed that collagen level was significantly decreased in combined treated group when compared with control . Results of present study suggest that P08253 and P14780 inhibition in presence of P36551 inhibitor prevents the development of experimental diabetic nephropathy in rats and can be a potential approach for the treatment . DB01017 effects on cerebral edema : relations with inflammatory and oxidative stress markers following traumatic brain injury in mice . One of the severe complications following traumatic brain injury ( TBI ) is cerebral edema and its effective treatment is of great interest to prevent further brain damage . This study investigated the effects of minocycline , known for its anti-inflammatory properties , on cerebral edema and its respective inflammatory markers by comparing different dose regimens , on oxidative stress and on neurological dysfunction following TBI . The weight drop model was used to induce TBI in mice . The brain water content was measured to evaluate cerebral edema . Inflammatory markers were detected by ELISA ( IL-1beta ) , zymography and Western blot ( P14780 ) . The oxidative stress marker ( glutathione levels ) and neurological function were measured by Griffith technique and string test , respectively . DB01017 was administered i.p. once ( 5 min ) , twice ( 5 min and 3 h ) or triple ( 5 min , 3 h and 9 h ) following TBI . The first dose of minocycline only varied ( 45 or 90 mg/kg ) , whereas the following doses were all at 45 mg/kg . The single and double administrations of minocycline reduced the increase of inflammatory markers at 6 h post-TBI . DB01017 also reduced cerebral edema at this time point , only after double administration and at the high dose regimen , although with no effect on the TBI-induced oxidized glutathione increase . The anti-edematous effect of minocycline persisted up to 24 h , upon a triple administration , and accompanied by a neurological recovery . In conclusion , we reported an anti-edematous effect of minocycline after TBI in mice according to a specific treatment regimen . These findings emphasize that the beneficial effects of minocycline depend on the treatment regimen following a brain injury . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema .

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Immunohistochemical detection of alpha1E voltage-gated Ca(2+) channel isoforms in cerebellum , P01308 -1 cells , and neuroendocrine cells of the digestive system . Polyclonal antibodies were raised against a common and a specific epitope present only in longer alpha1E isoforms of voltage-gated Ca(2+) channels , yielding an " anti-E-com " and an " anti-E-spec " serum , respectively . The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected P29320 -293 cells or membrane proteins derived from them . Cells from the insulinoma cell line P01308 -1 , tissue sections from cerebellum , and representative regions of gastrointestinal tract were stained immunocytochemically . P01308 -1 cells expressed an alpha1E splice variant with a longer carboxy terminus , the so-called alpha1Ee isoform . Similarily , in rat cerebellum , which was used as a reference system , the anti-E-spec serum stained somata and dendrites of Purkinje cells . Only faint staining was seen throughout the cerebellar granule cell layer . After prolonged incubation times , neurons of the molecular layer were stained by anti-E-com , suggesting that a shorter alpha1E isoform is expressed at a lower protein density . In human gastrointestinal tract , endocrine cells of the antral mucosa ( stomach ) , small and large intestine , and islets of Langerhans were stained by the anti-E-spec serum . In addition , staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae . We conclude that the longer alpha1Ee isoform is expressed in neuroendocrine cells of the digestive system and that , in pancreas , alpha1Ee expression is restricted to the neuroendocrine part , the islets of Langerhans. alpha1E therefore appears to be a common voltage-gated Ca(2+) channel linked to neuroendocrine and related systems of the body . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . P62937 modulates the sensitivity of HIV-1 to host restriction factors . Many mammalian species express restriction factors that confer host resistance to retroviral infection . Here we show that HIV-1 sensitivity to restriction factors is modulated by cyclophilin A ( CypA ) , a host cell protein that binds the HIV-1 capsid protein ( CA ) . In certain nonhuman primate cells , the CA-CypA interaction is essential for restriction : HIV-1 infectivity is increased > 100-fold by cyclosporin A ( DB00091 ) , a competitive inhibitor of the interaction , or by an HIV-1 CA mutation that disrupts CypA binding . Conversely , disruption of CA-CypA interaction in human cells reveals that CypA protects HIV-1 from the Ref-1 restriction factor . These findings suggest that HIV-1 has co-opted a host cell protein to counteract restriction factors expressed by human cells and that this adaptation can confer sensitivity to restriction in unnatural hosts . Manipulation of HIV-1 CA recognition by restriction factors promises to advance animal models and new therapeutic strategies for HIV-1 and AIDS . Contribution of P62937 to determination of simian immunodeficiency virus tropism : a progress update . An understanding of cellular factors that affect viral replication contributes to elucidation of the mechanism for the determination of viral tropism . P62937 ( CypA ) , a peptidyl-prolyl cis-trans isomerase ( PPIase ) , is an essential host factor for the efficient replication of human immunodeficiency virus type 1 ( HIV-1 ) in human cells . However , its role in simian immunodeficiency virus ( SIV ) replication has not been determined . In the 2008 US-Japan AIDS panel meeting , I have presented the effect of cyclosporine A ( DB00091 ) , a PPIase inhibitor , on replication of wild-type SIV . Interestingly , DB00091 treatment enhanced SIV replication in human cells but abrogated SIV replication in macaque cells , implying a species-specific effect of DB00091 on SIV replication . After this meeting , analysis using CypA knocked-down human cells indicated that CypA was considered inhibitory for SIV replication . These results suggest possible involvement of CypA in the determination of SIV tropism . P62937 is an inflammatory mediator that promotes atherosclerosis in apolipoprotein E-deficient mice . P62937 ( CyPA ; encoded by Ppia ) is a ubiquitously expressed protein secreted in response to inflammatory stimuli . CyPA stimulates vascular smooth muscle cell migration and proliferation , endothelial cell adhesion molecule expression , and inflammatory cell chemotaxis . Given these activities , we hypothesized that CyPA would promote atherosclerosis . P02649 -deficient ( Apoe(-/-) ) mice fed a high-cholesterol diet for 16 wk developed more severe atherosclerosis compared with Apoe(-/-)Ppia(-/-) mice . Moreover , CyPA deficiency was associated with decreased low-density lipoprotein uptake , P19320 ( vascular cell adhesion molecule 1 ) expression , apoptosis , and increased P29474 ( endothelial nitric oxide synthase ) expression . To understand the vascular role of CyPA in atherosclerosis development , bone marrow ( BM ) cell transplantation was performed . Atherosclerosis was greater in Apoe(-/-) mice compared with Apoe(-/-)Ppia(-/-) mice after reconstitution with CyPA(+/+) BM cells , indicating that vascular-derived CyPA plays a crucial role in the progression of atherosclerosis . These data define a role for CyPA in atherosclerosis and suggest CyPA as a target for cardiovascular therapies . DB00091 and sanglifehrin A enhance chemotherapeutic effect of cisplatin in P13671 glioma cells . Glioma is the most common type of brain tumors in adults , and treatment of high-grade gliomas is still palliative . Studies to date have revealed only modest effect in attenuating growth of these tumors with single agent therapy , but combination treatment appears to be more effective . P62937 ( CypA ) , a target of immunosuppressive drugs cyclosporin A ( DB00091 ) and sanglifehrin A ( SFA ) , is an intracellular protein that has peptidyl-prolyl cis-trans isomerase ( PPIase ) enzymatic activity . Previously , we showed that overexpressed CypA induced chemoresistance in cancer cells . Here we provide evidence that combination of cisplatin with either DB00091 or SFA synergistically enhances apoptotic cell death in P13671 glioma cells , compared with single agent treatment . Enhanced apoptotic cell death is a result of an increase in ROS generation and a decrease in intracellular glutathione levels . Consistently , CypA knockdown by siRNA also enhances cisplatin-induced apoptosis . Immunohistochemical analysis showed increased expression of CypA in human glioblastoma multiforme , but not in normal human astrocytes . CypA was also shown to be up-regulated in P13671 glioma cells during hypoxia . In conclusion , DB00091 or SFA in combination with cisplatin synergistically enhances cisplatin-induced apoptosis in P13671 glioma cells via inhibition of PPIase activity of CypA , indicating that development of new drugs that selectively inhibit the CypA PPIase activity without immune suppression may facilitate alleviation of chemoresistance in treatment of high-grade glioma . A cyclophilin A inducible expressed in gonad of zhikong scallop Chlamys farreri . P62937 ( CypA ) , a receptor for the immunosuppressive agent cyclosporin A ( DB00091 ) , is a cis-trans peptidyl-prolyl isomerase ( PPIase ) which accelerates the cis-trans isomerization of prolyl-peptide bonds , interacts with a variety of proteins and therefore regulates their activities . One CypA ( designated CfCypA ) cDNA was cloned from Chlamys farreri by expressed sequence tag ( EST ) and rapid amplification of cDNA ends ( RACE ) techniques . The full-length cDNA of CfCypA consisted of 1,248 nucleotides with a canonical polyadenylation signal sequence AATAAA , a poly ( A ) tail , and an open reading frame ( ORF ) of 495 nucleotides encoding a polypeptide of 164 amino acids . The deduced amino acid sequence shared high similarity with CypA from the other species , indicating that CfCypA should be a new member of the CypA family . Quantitative real-time ( RT ) PCR was employed to assess the mRNA expression of CfCypA in various tissues and its temporal expression in haemocytes and gonad of scallops challenged with Vibrio anguillarum . The mRNA transcripts of CfCypA could be detected in all the examined tissues with highest expression level in gonad . After bacterial challenge , the expression level of CfCypA was almost unchanged in haemocytes , but up-regulated in gonad and increased to the peak ( 22.59-fold ; P < 0.05 ) at 4 h post-injection , and then dropped to the original level at 8 h post-injection . These results indicated that CfCypA was constitutive expressed in haemocytes , but could be induced in gonad , and perhaps played a critical role in response to the bacterial challenge in gonad . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Knockdown endogenous CypA with siRNA in U2OS cells results in disruption of F-actin structure and alters tumor phenotype . P62937 ( CypA ) was originally identified as a cytosolic protein possessing peptidyl-prolyl isomerase activity . CypA has been shown to play a pivotal role in the immune response , but little is known about other molecular mechanisms of CypA-mediated biologic events . In our present study , we demonstrate that knockdown CypA expression using RNAi in U2OS cells resulted in disruption of the F-actin structure , as well as decreased anchorage-independent growth , proliferation , and migration . Wild-type U2OS cells treated with cyclosporine A ( DB00091 ) , a peptidyl-prolyl isomerase inhibitor , displayed the same phenotype as knockdown CypA cells , suggesting that the isomerase activity of CypA is required to maintain a normal phenotype . In vitro and in vivo binding assays revealed that CypA binds to O00401 , which functions in the nucleation of actin via the Arp2/3 complex . Pulse-chase labeling study indicated an enhanced degradation of O00401 in cell lacking CypA , suggesting that CypA is required for stabilizing O00401 to form a O00401 /Arp2/3 complex for the nucleation/initiation of F-actin polymerization . Marine invertebrates cross phyla comparisons reveal highly conserved immune machinery . Naturally occurring histocompatibility responses , following tissue-to-tissue allogeneic contacts , are common among numerous colonial marine invertebrate taxa , including sponges , cnidarians , bryozoans and ascidians . These responses , often culminating in either tissue fusions or rejections , activate a wide array of innate immune components . By comparing two allorejection EST libraries , developed from alloincompatible challenged colonies of the stony coral Stylophora pistillata and the ascidian Botryllus schlosseri , we revealed a common basis for innate immunity in these two evolutionary distant species . Two prominent genes within this common basis were the immunophilins , P62937 ( CypA ) and FK506-binding protein ( FKBP ) . In situ hybridizations revealed that mRNA expression of the coral and ascidian immunophilins was restricted to specific allorecognition effector cell populations ( nematoblasts and nematocytes in the coral and morula cells in the ascidian ) . The expressions were limited to only some of the effector cells within a population , disclosing disparities in numbers and location between naïve colonies and their immune challenged counterparts . Administration of the immunosuppression drug DB00091 -A during ascidian 's allogeneic assays inhibited both fusion and rejection reactions , probably through the inhibition of ascidian 's immunocytes ( morula cells ) movement and activation . Our results , together with previous published data , depict an immunophilins-based immune mechanism , which is similarly activated in allogeneic responses of distantly related animals from sponges to humans . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . P14735 binds to the nonglycosylated precursor of varicella-zoster virus gE protein found in the endoplasmic reticulum . P01308 degradation enzyme ( P14735 ) is a 110-kDa zinc metalloprotease found in the cytosol of all cells . P14735 degrades insulin and a variety of small proteins including amyloid-beta . Recently , P14735 has been proposed as the receptor for varicella-zoster virus ( VZV ) attachment . During our reassessment , some of the original studies were repeated and expanded in scope . We first confirmed that P14735 antibody reduced VZV spread . For additional controls , we repeated the same experiments with herpes simplex virus ( HSV ) -infected cells as well as uninfected cells . There was a visible reduction in HSV spread but less than seen in the VZV system . Of greater importance , P14735 antibody also inhibited the growth of uninfected cells . Second , we repeated the coprecipitation assays . We confirmed that antibodies to VZV gE ( open reading frame 68 ) coprecipitated P14735 and that anti- P14735 antibody coprecipitated gE . However , the detected gE protein was not the mature 98-kDa form ; rather , it was a precursor 73-kDa gE form found in the endoplasmic reticulum . Additional control experiments included VZV-infected cell cultures treated with tunicamycin to block gE glycosylation in the endoplasmic reticulum ; again , the anti- P14735 antibody coprecipitated a 73-kDa gE product . Finally , Orbitrap mass spectrometry analysis of a chromatographically purified gE sample revealed four cellular proteins associated with the unfolded protein response : P11021 ( P11021 ) , P11142 , P10809 , and P62937 ( peptidyl-propyl cis-trans isomerase ) . We conclude that P14735 protease binds to the 73-kDa gE precursor and that this event occurs in the cytosol but not as a receptor/ligand interaction . Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . P62937 is required for efficient human cytomegalovirus DNA replication and reactivation . Human cytomegalovirus ( HCMV ) is a large DNA virus belonging to the subfamily Betaherpesvirinae . Haematopoietic cells of the myeloid lineage have been shown to harbour latent HCMV . However , following terminal differentiation of these cells , virus is reactivated , and in an immunocompromised host acute infection can occur . It is currently unknown which viral and cellular factors are involved in regulating the switch between lytic and latent infections . P62937 ( CyPA ) is a cellular protein that acts as a major factor in virus replication and/or virion maturation for a number of different viruses , including human immunodeficiency virus , hepatitis C virus , murine cytomegalovirus , influenza A virus and vaccinia virus . This study investigated the role of CyPA during HCMV infection . CyPA expression was silenced in human foreskin fibroblast ( HF ) and THP-1 cells using small interfering RNA ( siRNA ) technology , or the cells were treated with cyclosporin A ( DB00091 ) to inhibit CyPA activity . Silencing CyPA in HF cells with siRNA resulted in an overall reduction in virus production characterized by delayed expression of immediate-early ( IE ) proteins , decreased viral DNA loads and reduced titres . Furthermore , silencing of CyPA in THP-1 cells pre- and post-differentiation prevented IE protein expression and virus reactivation from a non-productive state . Interestingly , it was observed that treatment of THP-1 cells with DB00091 prevented the cells from establishing a fully latent infection . In summary , these results demonstrate that CyPA expression is an important factor in HCMV IE protein expression and virus production in lytically infected HF cells , and is a major component in virus reactivation from infected THP-1 cells . Critical selection of internal control genes for quantitative real-time RT-PCR studies in lipopolysaccharide-stimulated human THP-1 and K562 cells . The choice of internal control genes is important since it may affect the study outcome in RT-qPCR . Indeed , it is well-known that expression levels of traditional internal control genes can vary across tissue types and across experimental settings within one specific tissue type . The aim of this study is an evaluation of a set of housekeeping genes ( HKGs ) to be used in the normalization of gene expression in vitro different cultured cells , THP-1 and K562 . The transcriptional stability of eleven potential internal control genes ( P61513 , P60709 , P04406 , B(2)M , P23284 , P00558 , P62937 , P31040 , P20226 , P00492 and P40429 ) were evaluated using RT-qPCR and were compared in different treatment , that was un-stimulated or LPS-stimulated cells . The raw Ct values were determined for each candidate gene at different time points following LPS-stimulated or unstimulated cells . Furthermore , all data were analyzed by the geNorm , BestKeeper , and NormFinder validation programs . Results indicated that P23284 and P00558 were the most stable internal control genes in this study . P40429 was found to be the least stable . This study provides the comprehensive reported assessment of internal control genes for use in expression studies in vitro cultured cells . These findings further emphasize the need to accurately validate candidate internal control genes in the study before use in gene expression studies using RT-qPCR . Farnesylthiosalicylic acid inhibits mammalian target of rapamycin ( P42345 ) activity both in cells and in vitro by promoting dissociation of the P42345 -raptor complex . The mammalian target of rapamycin ( P42345 ) functions with raptor and Q9BVC4 in a signaling complex that controls rates of cell growth and proliferation . Recent results indicate that an inhibitor of the Ras signaling pathway , farnesylthiosalicylic acid ( Q9H8T0 ) , decreased phosphorylation of the P42345 effectors , Q13541 and P23443 , in breast cancer cells . Here we show that incubating 293T cells with Q9H8T0 produced a stable change in P42345 activity that could be measured in immune complex kinase assays using purified Q13541 as substrate . Similarly , Q9H8T0 decreased the Q13541 kinase activity of P42345 when added to cell extracts or to immune complexes containing P42345 . Incubating either cells or extracts with Q9H8T0 also decreased the amount of raptor that coimmunoprecipitated with P42345 , although having relatively little effect on the amount of Q9BVC4 that coimmunoprecipitated . The concentration effect curves of Q9H8T0 for inhibition of P42345 activity and for dissociation of the raptor- P42345 complex were almost identical . Caffeine , wortmannin , LY294002 , and rapamycin- P62942 also markedly inhibited P42345 activity in vitro , but unlike Q9H8T0 , none of the other P42345 inhibitors appreciably changed the amount of raptor associated with P42345 . Thus , our findings indicate that Q9H8T0 represents a new type of P42345 inhibitor , which acts by dissociating the functional P42345 -raptor signaling complex . DB00091 regulates the levels of cyclophilin A in neuroblastoma cells in culture . P62937 ( CyP-A ) , a member of a highly conserved family of proteins , immunophilins , is the major intracellular receptor for the immunosuppressive drug , cyclosporin A ( DB00091 ) . CyP-A is widely expressed in many tissues , but is found in the highest concentration in brain tissues and may perform critical neuronal functions . DB00091 is a known neurotoxin . Therefore , understanding the regulation of CyP-A levels in nerve cells , particularly by DB00091 , is important . We have utilized murine neuroblastoma ( NB ) cells as an experimental model to investigate this issue . Our results show that DB00091 alone was sufficient to induce morphological differentiation in undifferentiated NB cells and to increase CyP-A levels as determined by immunostaining . However , inducing terminal differentiation by elevating adenosine 3',5'-cyclic monophosphate ( DB02527 ) levels using either 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone ( RO20-1724 ) , an inhibitor of cyclic nucleotide phosphodiesterase , or prostaglandin E1 ( PGE1 ) , a stimulator of adenylate cyclase , was not sufficient to increase CyP-A levels . DB00091 was required to increase CyP-A levels in both RO20-1724- and PGE1-induced differentiated NB cells . Increases in CyP-A levels , however , occurred without any change in the expression of the CyP-A gene as determined by reverse-transcriptase polymerase-chain reaction analysis using ( CyP-A ) -specific primers . These results suggest that DB00091 regulates the level of its own binding protein , CyP-A , in both undifferentiated and DB02527 -induced differentiated NB cells in culture . Transgenic mice overexpressing cyclophilin A are resistant to cyclosporin A-induced nephrotoxicity via peptidyl-prolyl cis-trans isomerase activity . DB00091 ( DB00091 ) suppresses immune reaction by inhibiting calcineurin activity after forming complex with cyclophilins and is currently widely used as an immunosuppressive drug . P62937 ( CypA ) is the most abundantly and ubiquitously expressed family member of cyclophilins . We previously showed that DB00091 toxicity is mediated by ROS generation as well as by inhibition of peptidyl-prolyl cis-trans isomerase ( PPIase ) activity of CypA in DB00091 -treated myoblasts [ FASEB J. 16 ( 2002 ) 1633 ] . Since DB00091 -induced nephrotoxicity is the most significant adverse effect in its clinical utilization , we here investigated the role of DB00091 inhibition of CypA PPIase activity in its nephrotoxicity using transgenic mouse models . Transgenic mice of either wild type ( CypA/wt ) or R55A PPIase mutant type ( CypA/R55A ) , a dominant negative mutant of CypA PPIase activity , showed normal growth without any apparent abnormalities . However , DB00091 -induced nephrotoxicity was virtually suppressed in CypA/wt mice , but exacerbated in CypA/R55A mice , compared to that of littermates . Also , life expectancy was extended in CypA/wt mice and shortened in CypA/R55A mice during DB00091 administration . Besides , DB00091 -induced nephrotoxicity was inversely related to the levels of catalase expression and activity . In conclusion , our data provide in vivo evidence that supplement of CypA PPIase activity allows animal 's resistance toward DB00091 -induced nephrotoxicity . Host cell species-specific effect of cyclosporine A on simian immunodeficiency virus replication . BACKGROUND : An understanding of host cell factors that affect viral replication contributes to elucidation of the mechanism for determination of viral tropism . P62937 ( CypA ) , a peptidyl-prolyl cis-trans isomerase ( PPIase ) , is a host factor essential for efficient replication of human immunodeficiency virus type 1 ( HIV-1 ) in human cells . However , the role of cyclophilins in simian immunodeficiency virus ( SIV ) replication has not been determined . In the present study , we examined the effect of cyclosporine A ( DB00091 ) , a PPIase inhibitor , on SIV replication . RESULTS : SIV replication in human CEM-SS T cells was not inhibited but rather enhanced by treatment with DB00091 , which inhibited HIV-1 replication . DB00091 treatment of target human cells enhanced an early step of SIV replication . CypA overexpression enhanced the early phase of HIV-1 but not SIV replication , while CypA knock-down resulted in suppression of HIV-1 but not SIV replication in CEM-SS cells , partially explaining different sensitivities of HIV-1 and SIV replication to DB00091 treatment . In contrast , DB00091 treatment inhibited SIV replication in macaque T cells ; DB00091 treatment of either virus producer or target cells resulted in suppression of SIV replication . SIV infection was enhanced by CypA overexpression in macaque target cells . CONCLUSIONS : DB00091 treatment enhanced SIV replication in human T cells but abrogated SIV replication in macaque T cells , implying a host cell species-specific effect of DB00091 on SIV replication . Further analyses indicated a positive effect of CypA on SIV infection into macaque but not into human T cells . These results suggest possible contribution of CypA to the determination of SIV tropism . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . P62937 as a target of DB00515 chemosensitizers . Platinum-based chemotherapeutics are the mainstay of treatment of a range of tumors achieving high response rates but limited in the course of disease by appearance of drug resistance . Tumor cells respond with reduced uptake and increased intracellular inactivation of the drugs , as well as increased DNA repair and general resistance to chemotherapyinduced cell death . DB00515 is known to induce expression of cyclophilins , a group of proteins that have peptidyl-prolyl cis-trans isomerase ( PPIase ) and molecular chaperone activities , as stress response . P62937 ( CypA ) and other members of this family are inhibited by cyclosporin A ( DB00091 ) which sensitized diverse drug-resistant tumor cell lines in vitro to cisplatin . This effect of DB00091 was attributed to metabolic changes , inhibition of DNA repair , enhancement of apoptosis , altered intracellular signal transduction or increased production of reactive oxygen species ( ROS ) , although no definitive explanation was provided so far . Several clinical trials employing cisplatin/carboplatin in combination with DB00091 yielded unsatisfactory results . Since viral replication was found to be dependent on cyclophilins of the host cells , effective new inhibitors , different from DB00091 or with low or absent immunosuppressive activity , are in development or clinical trials . Sanglifehrins are more potent than DB00091 and proved to increase toxicity of cisplatin against hepatocellular cancer cells in vitro . These novel cyclophilin inhibitors may offer new opportunities to achieve reversal of resistance to platinumbased drugs in refractory patients . Responsive cancer patients may be enriched in clinical trials by an identification of the downstream targets of Cyps responsible for chemoresistance . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . HCV NS5A and Q00978 compete for CypA binding . BACKGROUND & AIMS : P62937 ( CypA ) is vital for HCV replication . Cyp inhibitors successfully decrease viral loads in HCV-infected patients . However , their mechanisms of action remain unknown . Since interferon ( IFN ) can also suppress HCV replication , we asked whether a link between CypA and the IFN response exists . METHODS : We used cellular and recombinant pulldown approaches to investigate the possibility of a specific association of CypA with host ligands . RESULTS : We found for the first time that CypA binds to a major component of the IFN response - the IFN regulatory factor 9 ( Q00978 ) . Q00978 is the DNA-binding component of the transcriptional IFN-stimulated gene factor 3 ( ISGF3 ) . CypA binds directly to Q00978 via its peptidyl-prolyl isomerase ( PPIase ) pocket . Cyp inhibitors such as cyclosporine A ( DB00091 ) or non-immunosuppressive derivates such as alisporivir and SCY-635 , prevent Q00978 -CypA complex formation . CypA binds to the C-terminal Q969Q1 -association-domain ( IAD ) , but not to the DNA-binding or linker domains of Q00978 . Remarkably , CypA associates with the multimeric ISGF3 complex . We also obtained evidence that CypA neutralization enhances IFN-induced transcription . Interestingly , the hepatitis C virus ( HCV ) non-structural 5A ( NS5A ) protein , which is known to modulate the IFN response , competes with Q00978 for CypA binding and can prevent the formation of Q00978 -CypA complexes . CONCLUSIONS : This study demonstrates for the first time that CypA binds specifically to a component of the Janus kinase/signal transducer and activator of transcription ( JAK/ P35610 ) pathway , Q00978 . This study also reveals a novel opportunity of HCV to modulate the IFN response via NS5A . In utero activation of fetal memory T cells alters host regulatory gene expression and affects HIV susceptibility . In utero priming to malaria antigens renders cord blood mononuclear cells ( CBMC ) more susceptible to productive HIV infection in vitro in the absence of exogenous stimulation . This provides a unique model to better understand mechanisms affecting lymphocyte susceptibility to HIV infection in vivo . Effector memory CD3(+) P01730 (+) T cells ( T(EM) ) were the exclusive initial targets of HIV with rapid spread to central memory cells . HIV susceptibility correlated with increased expression of CD25 and HLA-DR on T(EM) . Virus entered all samples equally , however gag/pol RNA was only detected in HIV susceptible samples , suggesting regulation of proviral gene transcription . Targeted analysis of human genes in memory T cells showed greater expression of P01579 , O95644 , P10914 , P01100 , and P62937 and decreased expression P25490 and Q12800 in HIV susceptible samples . Thus fetal priming to exogenous antigens enhances specific proviral gene transcription pathways in effector memory cells that may increase risk of vertical transmission of HIV . Immunophilins in nervous system degeneration and regeneration . Immunophilins are receptors for immunosuppressive drugs like cyclosporin A , FK506 , rapamycin and their non- immunosuppressive analogs , which are collectively referred to as " immunophilin ligands " ( Q53GA4 ) . DB00091 binds to a class of IP called cyclophilins , whereas the receptors for FK506 and rapamycin belong to the family of FK506- binding proteins ( FKBP ) . The latter are designated according to their molecular weight : P62942 , 25 , 52 etc . FKBP levels in the rat brain are up to 50 times higher than in the immune system . P62942 is associated with IP3 and ryanodine receptors present on the endoplasmic reticulum and plays a role in stabilizing calcium release . It has also been proposed to be a modulator of the TGFbeta receptor activity . Crush injury of facial or sciatic nerves in rat leads to markedly increased P62942 levels in the respective nerve nuclei and this increase is related to nerve regeneration . P62937 protects cells from death following expression of mutant Cu/ Zn superoxide dismutase , which is associated with familial amyotrophic lateral sclerosis . Our recent studies show that P62942 and Q02790 are expressed in the human nervous system , especially in the substantia nigra- deep gray matter axis . In neurodegenerative diseases , P62942 levels increase in neurons situated in areas of pathology . This IP colocalizes with synaptophysin and alpha- synuclein , suggesting that it may become a novel marker of pathology . Immunophilins participate in axonal transport , synaptic vesicle assembly and may play a role in neuroprotection against abnormal protein aggregation , suggesting a potential avenue of therapeutic interventions . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . P62937 interacts with domain II of hepatitis C virus NS5A and stimulates RNA binding in an isomerase-dependent manner . NS5A plays a critical , yet poorly defined , role in hepatitis C virus genome replication . The protein consists of three domains , each of which is able to bind independently to the 3' untranslated region ( UTR ) of the viral positive strand genomic RNA . The peptidyl-prolyl isomerase cyclophilin A ( CypA ) binds to domain II , catalyzing cis-trans isomerization . CypA inhibitors such as cyclosporine ( DB00091 ) have been shown to inhibit hepatitis C virus ( HCV ) replication . We show here that CypA stimulated domain II RNA binding activity , and this stimulation was abrogated by DB00091 . An isomerase mutant of CypA ( H126Q ) failed to bind to domain II and did not stimulate RNA binding . Finally , we demonstrate that the RNA binding of two domain II mutants , the D316E and D316E/Y317N mutants , previously shown to exhibit CypA independence for RNA replication , was unaffected by CypA . This study provides an insight into the molecular mechanism of CypA activity during HCV replication and further validates the use of CypA inhibitors in HCV therapy . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Cyclophilin interactions with incoming human immunodeficiency virus type 1 capsids with opposing effects on infectivity in human cells . P62937 ( CypA ) is a peptidyl-prolyl isomerase that binds to the capsid protein ( CA ) of human immunodeficiency virus type 1 ( HIV-1 ) and by doing so facilitates HIV-1 replication . Although CypA is incorporated into HIV-1 virions by virtue of CypA-Gag interactions that occur during virion assembly , in this study we show that the CypA-CA interaction that occurs following the entry of the viral capsid into target cells is the major determinant of CypA 's effects on HIV-1 replication . Specifically , by using normal and CypA-deficient Jurkat cells , we demonstrate that the presence of CypA in the target and not the virus-producing cell enhances HIV-1 infectivity . Moreover , disruption of the CypA-CA interaction with cyclosporine A ( DB00091 ) inhibits HIV-1 infectivity only if the target cell expresses CypA . The effect of DB00091 on HIV-1 infection of human cells varies according to which particular cell line is used as a target , and CA mutations that confer DB00091 resistance and dependence exert their effects only if target cells , and not if virus-producing cells , are treated with DB00091 . The differential effects of DB00091 on HIV-1 infection in different human cells appear not to be caused by polymorphisms in the recently described retrovirus restriction factor TRIM5alpha . We speculate that CypA and/or CypA-related proteins affect the fate of incoming HIV-1 capsid either directly or by modulating interactions with unidentified host cell factors . Induction of G2 arrest and binding to cyclophilin A are independent phenotypes of human immunodeficiency virus type 1 Vpr . P62937 ( CypA ) is a member of a family of cellular proteins that share a peptidyl prolyl cis-trans isomerase ( PPIase ) activity . CypA was previously reported to be required for the biochemical stability and function ( specifically , induction of G2 arrest ) of the human immunodeficiency virus type 1 ( HIV-1 ) protein R ( Vpr ) . In the present study , we examine the role of the Vpr-CypA interaction on Vpr-induced G2 arrest . We find that Vpr coimmunoprecipitates with CypA and that this interaction is disrupted by substitution of proline-35 of Vpr as well as incubation with the CypA inhibitor cyclosporine A ( DB00091 ) . Surprisingly , the presence of CypA or its binding to Vpr is dispensable for the ability of Vpr to induce G2 arrest . Vpr expression in CypA-/- cells leads to induction of G2 arrest in a manner that is indistinguishable from that in CypA+ cells . DB00091 abolished CypA-Vpr binding but had no effect on induction of G2 arrest or Vpr steady-state levels . In view of these results , we propose that the interaction with CypA is independent of the ability of Vpr to induce cell cycle arrest . The interaction between Vpr and CypA is intriguing , and further studies should examine its potential effects on other functions of Vpr . UTP induces osteopontin expression through a coordinate action of NFkappaB , activator protein-1 , and upstream stimulatory factor in arterial smooth muscle cells . P10451 ( P10451 ) is an important chemokinetic agent for several cell types . Our earlier studies have shown that its expression is essential for uridine triphosphate ( UTP ) -mediated migration of vascular smooth muscle cells . We demonstrated previously that the activation of an AP-1 binding site located 76 bp upstream of the transcription start in the rat P10451 promoter is involved in the induction of P10451 expression . In this work , using a luciferase promoter deletion assay , we identified a new region of the rat P10451 promoter ( -1837 to -1757 ) that is responsive to UTP . This region contains an NFkappaB site located at -1800 and an Ebox located at -1768 . Supershift electrophoretic mobility shift assay and chromatin immunoprecipitation assays identified NFkappaB and P22415 -1/ P22415 -2 as the DNA binding proteins induced by UTP , respectively , for these two sites . Using dominant negative mutants of O15111 and P22415 transcription factors , we confirmed that NFkappaB and P22415 -1/ P22415 -2 are involved in the UTP-mediated expression of P10451 . Using a pharmacological approach , we demonstrated that P22415 proteins are regulated by the extracellular signal-regulated kinase ( P29323 )1/2 pathway , just as the earlier discovered AP-1 complex , whereas NFkappaB is up-regulated through PKCdelta signals . Finally , our work suggests that the UTP-stimulated P10451 expression involves a coordinate regulation of PKCdelta-NFkappaB , P27361 /2- P22415 , and P27361 /2/NAD(P)H oxidase AP-1 signaling pathways . 1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea : a novel cyclophilin A allosteric activator . P62937 ( CypA ) plays an important role in many physiology processes and its overexpression has been involved in many diseases including immune disease , viral infection , neuro-degenerative disease , and cancer . However , the actual role of CypA in the diseases is still far from clear , and a complete understanding of CypA is necessary in order to direct more specific and effective therapeutic strategies . Based on the screening of our in-house library through the isomer-specific proteolysis method , we find a CypA activator ( 1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea ) , compound 1a , which can increase CypA 's PPIase activity and give allosteric behavior . The binding affinity of compound 1a to CypA has been confirmed by Fortebio 's Octet Q13123 system and the increased phosphorylation of P29323 in H446 cells is observed by treatment with both compound 1a and DB00091 . In order to further evaluate the binding mode between the activator and CypA , the allosteric binding site and allosteric mechanism of CypA are investigated by molecular dynamics ( MD ) simulations in combination with mutagenesis experiments . The results show that the allosteric binding site of CypA is 7Å away from its catalytic site and is composed of Cys52 , His70 , His54 , Lys151 , Thr152 and Lys155 . Compound 1a binds to the allosteric site of CypA , stabilizing the active conformation of catalytic residues , and finally promotes the catalytic efficiency of CypA . We believe our finding of the CypA allosteric activator will be used as an effective chemical tool for further studies of CypA mechanisms in diseases . Two cyclophilin A homologs with shared and distinct functions important for growth and virulence of Cryptococcus neoformans . P62937 is the target of the immunosuppressant cyclosporin A ( DB00091 ) and is encoded by a single unique gene conserved from yeast to humans . In the pathogenic fungus Cryptococcus neoformans , two homologous linked genes , P15085 and P48052 , were found to encode two conserved cyclophilin A proteins . In contrast to Saccharomyces cerevisiae , in which cyclophilin A mutations confer DB00091 resistance but few other phenotypes , cyclophilin A mutations conferred dramatic phenotypes in C. neoformans . The Cpa1 and Cpa2 cyclophilin A proteins play a shared role in cell growth , mating , virulence and DB00091 toxicity . The Cpa1 and Cpa2 proteins also have divergent functions . cpa1 mutants are inviable at 39 degrees C and attenuated for virulence , whereas cpa2 mutants are viable at 39 degrees C and fully virulent . cpa1 cpa2 double mutants exhibited synthetic defects in growth and virulence . P62937 active site mutants restored growth of cpa1 cpa2 mutants at ambient but not at higher temperatures , suggesting that the prolyl isomerase activity of cyclophilin A has an in vivo function . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . Secreted cyclophilin A , a peptidylprolyl cis-trans isomerase , mediates matrix assembly of hensin , a protein implicated in epithelial differentiation . Q9UGM3 is a rabbit ortholog of Q9UGM3 , a multifunctional , multidomain protein implicated in the regulation of epithelial differentiation , innate immunity , and tumorigenesis . Q9UGM3 in the extracellular matrix ( Q13201 ) induced morphological changes characteristic of terminal differentiation in a clonal cell line ( clone C ) of rabbit kidney intercalated cells . Although hensin is secreted in monomeric and various oligomeric forms , only the polymerized Q13201 form is able to induce these phenotypic changes . Here we report that hensin secretion and matrix assembly were inhibited by the peptidylprolyl cis-trans isomerase ( PPIase ) inhibitors cyclosporin A ( DB00091 ) and a derivative of cyclosporin A with modifications in the d- DB00133 side chain ( Cs9 ) but not by the calcineurin pathway inhibitor FK506 . PPIase inhibition led to failure of hensin polymerization in the medium and Q13201 , plus the loss of apical cytoskeleton , apical microvilli , and the columnar epithelial shape of clone C cells . P62937 was produced and secreted into the media to a much greater extent than cyclophilins B and C . Our results also identified the direct DB00091 -sensitive interaction of cyclophilin A with hensin , suggesting that cyclophilin A is the PPIase that mediates the polymerization and matrix assembly of hensin . These results are significant because this is the first time a direct role of peptidylprolyl cis-trans isomerase activity has been implicated in the process of epithelial differentiation . P62937 -deficient mice are resistant to immunosuppression by cyclosporine . DB00091 is an immunosuppressive drug that is widely used to prevent organ transplant rejection . Known intracellular ligands for cyclosporine include the cyclophilins , a large family of phylogenetically conserved proteins that potentially regulate protein folding in cells . Immunosuppression by cyclosporine is thought to result from the formation of a drug-cyclophilin complex that binds to and inhibits calcineurin , a serine/threonine phosphatase that is activated by TCR engagement . Amino acids within the cyclophilins that are critical for binding to cyclosporine have been identified . Most of these residues are highly conserved within the 15 mammalian cyclophilins , suggesting that many are potential targets for the drug . We examined the effects of cyclosporine on immune cells and mice lacking Ppia , the gene encoding the prototypical cyclophilin protein cyclophilin A . TCR-induced proliferation and signal transduction by Ppia(-/-) P01730 (+) T cells were resistant to cyclosporine , an effect that was attributable to diminished calcineurin inhibition . Immunosuppressive doses of cyclosporine failed to block the responses of Ppia(-/-) mice to allogeneic challenge . Rag2(-/-) mice reconstituted with Ppia(-/-) splenocytes were also cyclosporine resistant , indicating that this property is intrinsic to Ppia(-/-) immune cells . Thus , among multiple potential ligands , CypA is the primary mediator of immunosuppression by cyclosporine . P62937 is upregulated in small cell lung cancer and activates P27361 /2 signal . P62937 ( CypA ) , a peptidyl-prolyl cis-trans isomerase ( PPIase ) , was originally identified as the intracellular receptor for cyclosporin A ( DB00091 ) . Recently , correlations of CypA with tumor pathogenesis have been studied . Here , we studied the expression of CypA and its receptor CD147 in several kinds of lung cancer cells as well as a normal lung cell and found that in H446 cell , a kind of small cell lung cancer cell , the expression are the highest . The exogeneous CypA protein can substantially stimulate H446 cell growth in dependence on its PPIase activity . We also showed that CypA protein can stimulate P27361 /2 signal in dose and time dependent manners and almost has no effect to p38 and JNK signals . Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for small cell lung cancer . P35372 expression is increased in inflammatory bowel diseases : implications for homeostatic intestinal inflammation . BACKGROUND AND AIMS : Recent studies with mu opioid receptor ( MOR ) deficient mice support a physiological anti-inflammatory effect of MOR at the colon interface . To better understand the potential pharmacological effect of certain opiates in inflammatory bowel diseases ( Q9UKU7 ) , we ( 1 ) evaluated the regulation in vivo and in vitro of human MOR expression by inflammation ; and ( 2 ) tested the potential anti-inflammatory function of a specific opiate ( DALDA ) in inflamed and resting human mucosa . PATIENTS AND METHODS : Expression of MOR mRNA and protein was evaluated in healthy and inflamed small bowel and colonic tissues , isolated peripheral blood mononuclear cells and purified monocytes , and P01730 + and CD8+ T cells from healthy donors and Q9UKU7 patients . The effect of cytokines and nuclear factor kappaB ( NFkappaB ) activation on MOR expression in lymphocyte T and monocytic human cell lines was assessed . Finally , DALDA induced anti-inflammatory effect was investigated in mucosal explants from controls and Q9UKU7 patients . RESULTS : MOR was expressed in ileal and colonic enteric neurones as well as in immunocytes such as myeloid cells and P01730 + and CD8+ T cells . Overexpressed in active Q9UKU7 mucosa , MOR was significantly enhanced by cytokines and repressed by NFkappaB inhibitor in myeloid and lymphocytic cell lines . Furthermore , ex vivo DALDA treatment dampened tumour necrosis factor alpha mRNA expression in the colon of active Q9UKU7 patients . CONCLUSIONS : Given the increased expression of MOR and the ex vivo beneficial effect of DALDA in active Q9UKU7 , natural and/or synthetic opioid agonists could help to prevent overt pathological intestinal inflammation . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Evidences of monomer , dimer and trimer of recombinant human cyclophilin A . P62937 ( CyPA ) is a cytosolic receptor of immunosuppressive drug cyclosporin A ( DB00091 ) which possesses peptidyl-prodyl cis/trans isomerase ( PPIase ) activity . The recombinant human CyPA ( rhCyPA ) gene has been expressed in E. coli M15 . Purification was performed using salting-out , as well as Sephacryl S-100 and DEAE-Sepharose CL-6B column chromatography . The molecular weight is about 18 kDa , confirmed by SDS-PAGE and mass spectrum . The results of Native-PAGE and immunoblotting showed the existence of three bands , which agreed well with the gel filtration results . The molecular mass of the three bands determined via CTAB gel electrophoresis and SDS-PAGE ( rhCyPA cross-linked with glutaraldehyde ) was 18 kDa , 36 kDa and 54 kDa respectively . Further more , the native rhCyPA and the cross-linked rhCyPA had the similar chromatographic behavior in gel filtration . All of the evidences above suggest that rhCyPA exists in forms of monomer , dimer and trimer . Moreover , we observed that even at low protein concentrations CyPA largely occurs as a dimer in solution , and enzyme kinetic parameters showed that activity of dimer was much higher than monomer or trimer , which probably have some biological significances . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) Genetic variants in epidermal growth factor receptor pathway genes and risk of esophageal squamous cell carcinoma and gastric cancer in a Chinese population . The epidermal growth factor receptor ( P00533 ) signaling pathway regulates cell proliferation , differentiation , and survival , and is frequently dysregulated in esophageal and gastric cancers . Few studies have comprehensively examined the association between germline genetic variants in the P00533 pathway and risk of esophageal and gastric cancers . Based on a genome-wide association study in a Han Chinese population , we examined 3443 SNPs in 127 genes in the P00533 pathway for 1942 esophageal squamous cell carcinomas ( ESCCs ) , 1758 gastric cancers ( GCs ) , and 2111 controls . SNP-level analyses were conducted using logistic regression models . We applied the resampling-based adaptive rank truncated product approach to determine the gene- and pathway-level associations . The P00533 pathway was significantly associated with GC risk ( P = 2.16×10(-3) ) . Gene-level analyses found 10 genes to be associated with GC , including P06241 , P45983 , P45985 , P08754 , Q02750 , Q9Y490 , P16471 , P16885 , Q9UBS0 , and Q92569 ( P < 0.05 ) . For ESCC , we did not observe a significant pathway-level association ( P = 0.72 ) , but gene-level analyses suggested associations between P08754 , Q04844 , O96013 , O00401 , and Q96J02 , and ESCC ( P < 0.05 ) . Our data suggest an association between specific genes in the P00533 signaling pathway and risk of GC and ESCC . Further studies are warranted to validate these associations and to investigate underlying mechanisms . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . P62937 , the major intracellular receptor for the immunosuppressant cyclosporin A , maps to chromosome 7p11.2-p13 : four pseudogenes map to chromosomes 3 , 10 , 14 , and 18 . P62937 ( CyP-A ) , the major intracellular receptor for the immunosuppressant cyclosporin A ( DB00091 ) , is a member of the immunophilin class of proteins , which all possess peptidyl-prolyl cis-trans isomerase activity and , therefore , are believed to be involved in protein folding and/or intracellular protein transport . The CyP-A protein is encoded by a single gene ; in addition , 15 pseudogenes have been identified . Recently , specific binding of CyP-A to the human immunodeficiency virus type 1 ( HIV-1 ) gag protein has been reported . Interestingly , this interaction can be inhibited by the immunosuppressant DB00091 and also by nonimmunosuppressive , CyP-A-binding DB00091 derivatives , which were also shown to exhibit potent anti-HIV-1 activity . Results thus indicate that CyP-A may have an essential function in HIV-1 replication . Using a panel of somatic rodent-human cell hybrids and PCR technology , we localized the coding cyclophilin A gene ( P62937 ) on chromosome 7 and four pseudogenes ( PPIP2 , PPIP3 , PPIP4 , and PPIP6 ) on chromosomes 14 , 10 , 18 , and 3 , respectively . Using chromosome 7 and chromosome 10 deletion hybrid panels , we were able to localize further the coding gene to the region 7p11.2-p13 , as confirmed by fluorescence in situ hybridization analysis , and one pseudogene ( PPIP3 ) to the region 10q11.2-q23 . This is the first report on the regional mapping of members of the CyP-A gene family . Design and synthesis of conformationally constrained cyclophilin inhibitors showing a cyclosporin-A phenotype in C. elegans . P62937 ( CypA ) is a member of the immunophilin family of proteins and receptor for the immunosuppressant drug cyclosporin A ( DB00091 ) . Here we describe the design and synthesis of a new class of small-molecule inhibitors for CypA that are based upon a dimedone template . Electrospray mass spectrometry is utilised as an initial screen to quantify the protein affinity of the ligands . Active inhibitors and fluorescently labelled derivatives are then used as chemical probes for investigating the biological role of cyclophilins in the nematode Caenorhabditis elegans . Nephrotoxicity of cyclosporin A and FK506 : inhibition of calcineurin phosphatase . DB00091 ( DB00091 ; 50 mg/kg ) and Fujimycine ( FK506 ; 5 mg/kg ) , but not the related macrolide immunosuppressant rapamycin ( 5 mg/kg ) , caused a reduction of glomerular filtration rate , degenerative changes of proximal tubular epithelium , and hypertrophy of the juxtaglomerular apparatus in male Wistar rats when given for 10 days . The molecular mechanisms of DB00091 and FK506 toxicity were investigated . P62937 and FK506-binding protein , the main intracytoplasmic receptors for DB00091 and FK506 , respectively , were each detected in renal tissue extract . In the kidney , high levels of immunoreactive and enzymatically active calcineurin were found which were inhibited by the immunosuppressants DB00091 and FK506 , but not by rapamycin . Finally , specific immunophilin-drug-calcineurin complexes formed only in the presence of DB00091 and FK506 , but not rapamycin . These results suggest that the nephrotoxic effects of DB00091 and FK506 is likely mediated through binding to renal immunophilin and inhibiting calcineurin phosphatase . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . P62937 is an essential cofactor for hepatitis C virus infection and the principal mediator of cyclosporine resistance in vitro . DB00091 ( DB00091 ) and its derivatives potently suppress hepatitis C virus ( HCV ) replication . Recently , DB00091 -resistant HCV replicons have been identified in vitro . We examined the dependence of the wild-type and DB00091 -resistant replicons on various cyclophilins for replication . A strong correlation between DB00091 resistance and reduced dependency on cyclophilin A ( CyPA ) for replication was identified . Silencing of CyPB or CyPC expression had no significant effect on replication , whereas various forms of small interfering RNA ( siRNA ) directed at CyPA inhibited HCV replication of wild-type but not DB00091 -resistant replicons . The efficiency of a particular siRNA in suppressing CyPA expression was correlated with its potency in inhibiting HCV replication , and expression of an siRNA-resistant CyPA cDNA rescued replication . In addition , an anti-CyPA antibody blocked replication of the wild-type but not the resistant replicon in an in vitro replication assay . Depletion of CyPA alone in the DB00091 -resistant replicon cells eliminated DB00091 resistance , indicating that CyPA is the chief mediator of the observed DB00091 resistance . The dependency on CyPA for replication was observed for both genotype ( GT ) 1a and 1b replicons as well as a GT 2a infectious virus . An interaction between CyPA and HCV RNA as well as the viral polymerase that is sensitive to DB00091 treatment in wild-type but not in resistant replicons was detected . These findings reveal the molecular mechanism of DB00091 resistance and identify CyPA as a critical cellular cofactor for HCV replication and infection . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . HIV protease substrate conformation : modulation by cyclophilin A . P62937 ( CyPA ) , a cytosolic peptidyl-prolyl trans-cis isomerase can accelerate the trans-cis isomerization of Xxx-Pro peptide bonds . One- and two-dimensional 1H-NMR spectroscopy were used to determine that the heptapeptide DB00133 -Gln- DB00174 - DB00135 -Pro- DB00167 - DB00161 , a model peptide of an HIV-1 protease cleavage site in the gag polyprotein of HIV-1 , is a substrate for CyPA . Experiments revealed a slow exchange about the DB00135 -Pro peptide bond with 30 +/- 5 % in the cis conformation ( pH 1-9 ) . While the interconversion rate is too slow to measure by kinetic NMR methods in the absence of CyPA , these methods , saturation transfer and NOE experiments , established that CyPA enhanced the rate of trans-cis interconversion , a process inhibited by cyclosporin A ( DB00091 ) . With a substrate:CyPA ratio of 40:1 , an interconversion rate of 2.5 s(-1) at 25 degrees C was observed . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . P62937 is required for Q9C035 {alpha}-mediated resistance to HIV-1 in Old World monkey cells . The peptidyl-prolyl isomerase cyclophilin A ( CypA ) embraces an exposed , proline-rich loop on HIV-1 capsid ( CA ) and renders reverse transcription complexes resistant to an antiviral activity in human cells . A CypA fusion with Q9C035 that is unique to New World owl monkeys also targets HIV-1 CA , but this interaction potently inhibits infection . A similar block to HIV-1 infection in Old World monkeys is attributable to the alpha isoform of the Q9C035 orthologue in these species . To determine whether HIV-1 restriction by Old World monkey TRIM5alpha is modulated by the CA-CypA interaction , RNA interference was used to disrupt CypA in cells from African green monkeys and rhesus macaques . HIV-1 infectivity increased in response to CypA knock-down to the same extent that it increased in response to Q9C035 knock-down . CypA knock-down eliminated the HIV-1 stimulatory effect of cyclosporin A ( DB00091 ) , a competitive inhibitor of the CypA-CA interaction , or of CA mutants that block binding to CypA but caused no change in titer of retroviruses that do n't interact with CypA . Simultaneous knock-down of both CypA and Q9C035 caused minimal additional increase in titer , suggesting that CypA inhibits HIV-1 replication in these cells because it is required for CA recognition by TRIM5alpha . Finally , DB00091 increased HIV-1 titer in otherwise nonrestrictive feline cells but only after these cells were transduced with Old World monkey TRIM5alpha . Thus , CypA is required for HIV-1 restriction by Old World monkey orthologues of TRIM5alpha . Differential expression of cyclophilin isoforms during keratinocyte differentiation . P62937 , the major intracellular binding protein for the immunosuppressive drug cyclosporin A ( DB00091 ) , was studied in human keratinocytes during differentiation both in vivo and in vitro . Analysis of cyclophilin by gel-filtration radiobinding-assay with tritiated DB00091 showed one specific radioactive peak at 17 kDa . By this technique , the levels of cyclophilin ( mean 55.23 +/- 8.43 pmol/mg protein ) did not significantly differ during keratinocyte differentiation . When the protein extracts from calcium-induced differentiating keratinocytes and normal human skin were analysed by PAGE radiobinding-assay , two specific radioactive DB00091 -binding peaks were detected . The major peak ( RF 0.13 ) was expressed in all samples ( mean 47.32 +/- 17.53 pmol/mg protein ) whereas the minor peak ( RF 0.23 ) was dramatically decreased about 6-fold in abnormally differentiated skin ( psoriasis ) as well as in non-differentiated keratinocytes . At least six [3H] DB00091 -binding isoforms with pI values ranging from 5.58 to 7.75 were detected by isoelectrofocusing autoradio-blotting-assay in normal human skin ; three of them immunoreacted with antibodies to cyclophilin . These results demonstrated the presence of several cyclophilin isoforms in human epidermal cells and an expression which correlated with the differentiation of human keratinocytes both in vivo and in vitro . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . SERCA2b activity is regulated by cyclophilins in human platelets . OBJECTIVE : The role of cyclophilins ( chaperones that are widely expressed in different cell types , including human platelets ) was explored in sarcoendoplasmic calcium ( Ca(2+) ) adenosine triphosphatase ( SERCA ) activity . METHODS AND RESULTS : Cyclophilin inhibition by cyclosporin A ( DB00091 ) evoked a time- and concentration-dependent reduction of Ca(2+) uptake by SERCA2b . However , other Ca(2+)-adenosine triphosphatases expressed in platelets , such as Q93084 and plasma membrane Ca(2+) adenosine triphophatase , remained unaltered after DB00091 treatment . Cypermethrin , a non- DB00091 -related calcineurin inhibitor , did not alter SERCA2b activity . Furthermore , SERCA2b was affected by other DB00091 analogues , which do not interfere with calcineurin , such as PKF-211-811-NX5 ( NIM811 ) and sanglifehrin A . Inhibition of the immunophilin family members using FK506 ( tacrolimus ) did not alter SERCA2b ability to sequester Ca(2+) into the dense tubular system . Coimmunoprecipitation experiments confirmed that cyclophilin A associates with SERCA2b and stromal interaction molecule-1 in resting platelets . This interaction is attenuated by the physiological agonist thrombin but enhanced by treatment with DB00091 or sanglifehrin A . CONCLUSIONS : P62937 is a regulator of SERCA2b in human platelets . P62937 and nuclear factor of activated T cells are essential in cyclosporine-mediated suppression of polyomavirus BK replication . Immunosuppressants have impacts on the development of polyomavirus-associated nephropathy . We previously demonstrated that cyclosporin A ( DB00091 ) suppressed polyomavirus BK ( BKV ) replication . The role of cyclophilin A ( CypA ) and nuclear factor of activated T cells ( NFAT ) in DB00091 -imposed suppression of BKV replication was determined in this study . Results demonstrated that knockdown of CypA but not CypB significantly reduced BKV large T antigen ( TAg ) expression and BKV titer . Overexpression of CypA reversed CypA siRNA-induced inhibition in BKV TAg expression . In addition , CypA overexpression attenuated the suppressive effect of DB00091 on TAg expression , suggesting CypA implicated in DB00091 -mediated anti-BKV effect . Knockdown of Q12968 abrogated TAg expression , while overexpression of Q12968 promoted TAg expression and augmented BKV promoter activity . Q12968 binding to the BKV promoter was verified by chromatin immunoprecipitation assay and electrophoretic mobility shift assay . Renal histology also displayed an increase in Q12968 expression in tubulointerstitium of BKV-associated nephropathy . Furthermore , overexpression of Q12968 rescued DB00091 -mediated inhibition of BKV load and TAg expression . A DB00091 analog , NIM811 , which can not block NFAT functionality , failed to suppress TAg expression . In conclusion , CypA and NFAT are indispensable in BKV replication . DB00091 inhibits BKV replication through CypA and NFAT , which may be potential targets of anti-BKV treatment .

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interacts_with DB00482?

multiple_choice

Quantum mechanics-based properties for 3D-QSAR . We have used a set of four local properties based on semiempirical molecular orbital calculations ( electron density ( ρ ) , hydrogen bond donor field ( HDF ) , hydrogen bond acceptor field ( P00748 ) , and molecular lipophilicity potential ( MLP ) ) for 3D-QSAR studies to overcome the limitations of the current force field-based molecular interaction fields ( MIFs ) . These properties can be calculated rapidly and are thus amenable to high-throughput industrial applications . Their statistical performance was compared with that of conventional 3D-QSAR approaches using nine data sets ( angiotensin converting enzyme inhibitors ( P12821 ) , acetylcholinesterase inhibitors ( AchE ) , benzodiazepine receptor ligands ( BZR ) , cyclooxygenase-2 inhibitors ( P35354 ) , dihydrofolate reductase inhibitors ( P00374 ) , glycogen phosphorylase b inhibitors ( GPB ) , thermolysin inhibitors ( THER ) , thrombin inhibitors ( THR ) , and serine protease factor Xa inhibitors ( fXa ) ) . The 3D-QSAR models generated were tested thoroughly for robustness and predictive ability . The average performance of the quantum mechanical molecular interaction field ( QM-MIF ) models for the nine data sets is better than that of the conventional force field-based MIFs . In the individual data sets , the QM-MIF models always perform better than , or as well as , the conventional approaches . It is particularly encouraging that the relative performance of the QM-MIF models improves in the external validation . In addition , the models generated showed statistical stability with respect to model building procedure variations such as grid spacing size and grid orientation . QM-MIF contour maps reproduce the features important for ligand binding for the example data set ( factor Xa inhibitors ) , demonstrating the intuitive chemical interpretability of QM-MIFs . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Quantification and characterisation of cyclo-oxygenase and lipid peroxidation inhibitory anthocyanins in fruits of Amelanchier . The levels of bioactive anthocyanins in the fruits of Amelanchier alnifolia , A. arborea and A. canadensis have been determined by HPLC . Cyanidin 3-galactoside ( 1 ) was present in the fresh fruit of the three species at concentrations of 155 , 390 and 165 mg/100 g , respectively . Cyanidin 3-glucoside ( 2 ) was present only in A. alnifolia and A. canadensis at concentrations of 54 and 48 mg/100 g , respectively . The anthocyanins were confirmed by LC- P19957 /MS and NMR studies . At 100 ppm , anthocyanin mixtures from the three species inhibited cyclo-oxygenase ( P36551 ) -1 and -2 enzymes at 66 and 67 % , 60 and 72 % , and 51 and 76 % , respectively . The positive controls used in the P36551 assays were aspirin , DB00482 and Vioxx at 180 , 1.67 and 1.67 ppm , respectively , and showed 74 and 69 % , 5 and 82 % and 0 and 85 % P23219 and P35354 inhibition , respectively . Anthocyanins 1 and 2 and cyanidin ( 3 ) inhibited P23219 enzyme 50.5 , 45.62 and 96.36 % , respectively , at 100 ppm , whereas P35354 inhibition was the highest for 3 at 75 % . In the lipid peroxidation inhibitory assay , anthocyanin mixtures at 10 ppm from the three species showed activities of 72 , 73 and 68 % , respectively , compared with 89 , 87 and 98 % for commercial anti-oxidants butylated hydoxyanisole , butylated hydroxytoluene , and tert-butylhydroxyquinone at 1.67 , 2.2 and 1.67 ppm , respectively . At 10 ppm , compounds 1-3 inhibited lipid peroxidation by 70 , 75 and 78 % , respectively . Detection of overexpressed P35354 in precancerous lesions of hamster pancreas and lungs by molecular imaging : implications for early diagnosis and prevention . The enzyme cyclooxygenase-2 ( P35354 ) is overexpressed in many cancers , cardiovascular disease , neurodegenerative disorders , and arthritis . Selective inhibitors of P35354 have been developed as therapeutics or preventive agents for these diseases . However , recent reports have revealed a significant increase in cardiovascular mortality in long-term users of the P35354 inhibitors Vioxx and DB00482 , emphasizing the need for noninvasive tests that allow the identification of individuals whose P35354 levels are overexpressed prior to assignment to treatment with these drugs . In this study , we have prepared a radioiodinated analogue of the selective P35354 inhibitor celecoxib , and verified its binding to the P35354 enzyme in vitro . Biodistribution studies in hamsters demonstrated significantly higher levels of radiotracer in animals treated with the tobacco carcinogen NNK in lung , pancreas , and liver . Assessment of P35354 levels by whole-body planar nuclear imaging two hours after injection of the radiotracer was suggestive of a distinct increase in P35354 in the pancreas and liver of a hamster treated for 10 weeks with NNK , in the lungs and liver of a second animal , and in the liver only , in two additional animals from the same treatment group . Immunostains showed selective overexpression of P35354 in pre-neoplastic lesions of the pancreas and lungs in only those animals that showed tracer accumulation in these organs and in the livers of all NNK-treated hamsters . Immunostains for P23219 yielded detectable reactions in the intestinal epithelium but not in pancreas , lungs , or liver , supporting the specificity of the tracer for P35354 . Our data provide proof of principle for the hypothesis that molecular imaging with radiolabeled P35354 inhibitors can be used for the noninvasive monitoring of overexpressed P35354 levels . Selective inhibition of P35354 is beneficial to mice infected intranasally with VSV . Cyclooxygenase ( P36551 ) is the key enzyme for prostaglandin ( PG ) synthesis . PGs are mediators of many critical physiological and inflammatory responses . There are two isoforms , P23219 and P35354 , both of which are constitutively expressed in the central nervous system ( CNS ) . Studies have shown that P23219 and P35354 are involved in physiological and pathological conditions of the brain . However , little is known about the role(s) of P36551 in the host defense system against a viral infection in the CNS . In this report , we used Vesicular Stomatitis Virus ( VSV ) induced acute encephalitis to distinguish between the contribution(s) of the two isoforms . P35354 activity was inhibited with a P35354 selective drug , celecoxib ( DB00482 ) , and P23219 was antagonized with SC560 . We found that inhibition of P35354 led to decreased viral titers , while P23219 antagonism did not have the same effect at day 1 post infection . 5-lipooxygenase ( P09917 ) expression and neutrophil recruitment in the CNS were increased in celecoxib-inhibited mice . Furthermore , mice treated with celecoxib expressed more DB00435 Synthase-1 ( Q8WY41 ) , a crucial component of the innate immune system in the restriction of VSV propagation . The expression of type 1 cytokines , P01579 and IL-12 , were also increased in celecoxib-treated mice . Acute generalized exanthematic pustulosis : a case and an overview of side effects affecting the skin caused by celecoxib and other P35354 inhibitors reported so far . A 55-year-old woman who was treated for periarthritis humeroscapularis with celecoxib ( DB00482 ) developed a generalized pustular exanthema on the head and upper trunk , accompanied by fever , leukocytosis and increased erythrocyte sedimentation rate . The histological findings were subcorneal pustules , necrotic keratinocytes , edema in the upper dermis and polymorphic perivascular infiltrates . Four days after stopping celecoxib , the pustules disappeared without any treatment . Four weeks after disappearance of the skin lesions , celecoxib demonstrated a positive lymphocyte stimulation test . In this article , we present to our knowledge the first case of acute generalized exanthematic pustulosis caused by celecoxib , and we give an overview of the side effects affecting the skin caused by celecoxib and other cyclooxygenase type 2 inhibitors reported so far . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Tolerability of selective cyclooxygenase inhibitor , celecoxib , in patients with analgesic intolerance . Intolerance reactions to acetyl salicylic acid ( ASA ) and nonsteroidal anti-inflammatory drugs ( NSAIDs ) are common and caused by inhibition of P23219 enzyme . Therefore , drugs that selectively inhibit P35354 enzyme may be safe in these subjects . In this study , we evaluated the tolerability of celecoxib , a selective P35354 inhibitor , in patients with analgesic intolerance . The eligible study population consisted of patients with a history of urticaria/angioedema , naso-ocular symptoms , bronchospasm , and/or anaphylactoid reaction induced by ASA and/or NSAIDs . A single-blind , placebo-controlled oral challenge test was performed in the hospital setting . On 2 separate days , 1/4 and 3/4 divided doses of placebo and celecoxib ( DB00482 200 mg , Pfizer , Turkey ) were given with 2-hour intervals . Seventy-five subjects ( mean age : 38.2 +/- 1.4 years ; F:M : 55:20 ) were included in the study . Twenty-one subjects had asthma . No reaction was observed with placebo or celecoxib provocation . Although celecoxib seems to be a safe alternative drug in our study group , considering its serious adverse events reported in the literature , the drug should be recommended for patients with analgesic intolerance only after being tested by an experienced allergist . The role of celecoxib in Rad51 expression and cell survival affected by gefitinib in human non-small cell lung cancer cells . Celecoxib ( DB00482 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor and gefitinib ( DB00317 (R) , ZD1839 ) is a selective epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor for human non-small cell lung cancer ( NSCLC ) . The addition of celecoxib to gefitinib to prolong the survival of patients with NSCLC still remains controversial and needs to be investigated . The Rad51 protein is essential for homologous recombination repair , and is overexpressed in chemo- or radioresistant carcinomas . In this study , we characterize the role of celecoxib in the cytotoxicity , P27361 /2 activation and Rad51 expression affected by gefitinib in NSCLC cells . We show that celecoxib can enhance the cytotoxicity induced by gefitinib in NSCLC cells . Treatment with celecoxib alone has no effect on the P27361 /2 activation , Rad51 mRNA and protein levels , however , combined treatment with gefitinib results in a significant reduction of phospho- P27361 /2 and Rad51 protein levels , and triggers the degradation of Rad51 via a 26S proteasome-dependent pathway . Expression of constitutively active Q02750 /2 vectors ( Q02750 /2-CA ) significantly rescues the decreased P27361 /2 activity , and restores Rad51 protein levels and cell survival under co-treatment with gefitinib and celecoxib . Furthermore , blocking P27361 /2 activation by U0126 ( Q02750 /2 inhibitor ) and knocking down Rad51 expression by transfection with small interfering RNA of Rad51 can enhance the cytotoxicity of celecoxib . DB00184 consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 - P30532 - P30926 cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5(-/-) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5*-nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake . Celecoxib and radiation therapy in non-small-cell lung cancer . Overexpression of cyclooxygenase-2 ( P35354 ) is frequently present in lung cancer and may play a significant role in carcinogenesis , invasion , and metastasis . It has been associated with shortened survival in patients with resected early-stage adenocarcinoma of the lung . P35354 inhibition decreases tumor cell proliferation in vivo and has been shown to enhance tumor radiosensitivity . Additionally , P35354 inhibition may protect normal pulmonary tissue from radiation fibrosis . Clinical studies are under way to assess the potential benefits and risks of P35354 inhibition in the treatment of lung cancer . The rationale for P35354 inhibitors in the treatment of lung cancer will be reviewed . The results of a phase II study assessing the acute toxicity of concurrent celecoxib ( DB00482 ) and thoracic irradiation in patients with non-small-cell lung cancer ( NSCLC ) are reported , and an ongoing Radiation Therapy Oncology Group study using celecoxib and concurrent radiation therapy for NSCLC in patients with intermediate prognostic factors is reviewed . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Selective P35354 inhibitors and the surgical patient . The identification of P35354 less than a decade ago has been followed by an unprecedented period of discovery and drug development ( Golden & Abramson , 1999 ) . Clinical studies thus far have established that the selective P35354 inhibitors , celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) are effective in the treatment of OA and RA , as are conventional NSAIDs . However , they do not cause the same adverse effects on the GI mucosa nor do they increase bleeding time as do conventional NSAIDs . Therefore , their use is not contraindicated in pre or postsurgical patients . Further investigation is underway to fully elucidate the role of P35354 and to determine any additional benefits of selective P35354 inhibition . Plasma levels of vascular endothelial growth factor during and after radiotherapy in combination with celecoxib in patients with advanced head and neck cancer . DB00482 and radiotherapy in advanced head and neck cancer . This phase I dose-escalation study seeks to determine the phase II recommended dose of cyclooxygenase type 2 ( P35354 ) inhibitor in patients with locally advanced squamous cell head and neck ( H & N ) cancer , treated with accelerated radiotherapy . Anti-vasculogenic effect of this treatment on serum vascular endothelial growth factor ( P15692 ) is examined . Patients were irradiated with curative intent ( 72Gy in 6weeks ) . Celecoxib was administered throughout the radiotherapy course . Serum P15692 level were tested during radiotherapy and in follow-up . Tumor specimens were stained to quantify the P35354 expression . Thirty-two patients completed the treatment . The dose of celecoxib was escalated ( 200 , 400 and 800mg bid , then de-escalated to 600mg bid ) . The acute toxicity related to the treatment in the first and second cohort did not reach grade III ; in the third cohort three patients had grade III radiation toxicity and one had celecoxib-related toxicity . In the last fourth cohort the toxicity was acceptable . Significant P15692 level drop ( p=0.011 ) was found between radiation day 1 and post-treatment visit . Significant decrease ( p=0.022 ) of the P15692 level was shown in patients with high P35354 expression in the tumor . Phase II recommended dose of celecoxib combined with accelerated radiotherapy in advanced H & N cancer was 600mg bid . A significant decrease of the post-treatment serum P15692 level compared to the initial level was noticed only in patients with high P35354 expression in tumors . P01116 , P00533 , P09619 -α , P10721 and P35354 status in carcinoma showing thymus-like elements ( CASTLE ) . BACKGROUND : CASTLE ( Carcinoma showing thymus-like elements ) is a rare malignant neoplasm of the thyroid resembling lymphoepithelioma-like and squamous cell carcinoma of the thymus with different biological behaviour and a better prognosis than anaplastic carcinoma of the thyroid . METHODS : We retrospectively investigated 6 cases of this very rare neoplasm in order to investigate the mutational status of P01116 , P00533 , P09619 -α and P10721 , as well as the immunohistochemical expression pattern of CD117 , P00533 and P35354 , and possibly find new therapeutic targets . RESULTS : Diagnosis was confirmed by a moderate to strong expression of P06127 , CD117 and CK5/6 , whereas thyroglobulin , calcitonin and Q15669 -1 were negative in all cases . Tumors were also positive for P35354 and in nearly all cases for P00533 . In four cases single nucleotide polymorphisms ( SNPs ) could be detected in exon 12 of the P09619 -α gene ( rs1873778 ) , in three cases SNPs were found in exon 20 of the P00533 gene ( rs1050171 ) . No mutations were found in the P10721 and P01116 gene . CONCLUSIONS : All tumors showed a P35354 expression as well as an P00533 expression except for one case and a wild-type P01116 status . No activating mutations in the P00533 , P10721 and P09619 -α gene could be detected . Our data may indicate a potential for targeted therapies , but if these therapeutic strategies are of benefit in CASTLE remains to be determined . VIRTUAL SLIDES : The virtual slide(s) for this article can be found here : http://www.diagnosticpathology.diagnomx.eu/vs/1658499296115016 . Approach to angiogenesis inhibition based on cyclooxygenase-2 . Two cyclooxygenase ( P36551 ) isoforms have been identified : P23219 and P35354 . P23219 is the constitutively expressed form of the enzyme and is ubiquitous in its distribution . P35354 is inducible and is present in inflammatory foci , tumors , and neovasculature . Expression of P35354 appears to be important in tumor promotion , growth , and metastasis . It is up-regulated in a variety of premalignant disorders and malignancies . P36551 inhibitors have a major role in the treatment of inflammation and pain . Epidemiologic evidence in patients who take nonsteroidal anti-inflammatory drugs links P36551 inhibition with decreases in malignant esophageal , stomach , colon , lung , and breast tumors . Nonselective P36551 inhibitors have demonstrated efficacy in control of familial adenomatous polyposis , a disorder associated with the development of thousands of benign intestinal polyps . The selective P35354 inhibitor celecoxib ( DB00482 , Pharmacia ) has been shown to reduce the number of adenomatous colorectal polyps in familial adenomatous polyposis as an adjunct to usual care . Celecoxib has recently been approved for this indication and offers the potential for equivalent or greater efficacy than that seen with nonselective P36551 inhibitors but without the gastrointestinal mucosal toxicity and the inhibition of platelet function associated with those agents . Angiogenesis is a feature of both benign and malignant disease . Because P35354 is up-regulated in the neovasculature of the rheumatoid pannus and in malignant tumors and their surrounding stroma , selective P35354 inhibitors may be able to modify the progression of these disorders through the control of angiogenesis . Membranous glomerulopathy and acute interstitial nephritis following treatment with celecoxib . Both membranous glomerulopathy and acute interstitial nephritis have been reported to occur following treatment with non-steroidal anti-inflammatory drugs . We report the first cases of membranous glomerulopathy and acute interstitial nephritis following treatment with celecoxib ( DB00482 ) , a selective P35354 inhibitor . The rapid and complete resolution of both conditions following discontinuation of DB00482 strongly implicates this agent in disease pathogenesis . These cases enlarge the spectrum of potential renal toxicities of the P35354 -specific non-steroidal anti-inflammatory drugs . Lipoperoxidation and cyclooxygenases 1 and 2 inhibitory compounds from Iryanthera juruensis . Plants from Iryanthera genus have been traditionally used as food supplements by South American Indians . The MeOH extract of leaves of Iryanthera juruensis , one of the plants endemic to the Amazon region and consumed in Brazil , and the hexane extract from its seeds inhibited lipid peroxidation ( P22079 ) and cyclooxygenase ( P23219 and -2 ) ) enzymes in in vitro assays . Further analyses of these extracts yielded 5-deoxyflavones ( 1-5 ) from the leaf extract and sargachromenol ( 6 ) , sargaquinoic acid ( 7 ) , a novel juruenolic acid ( 8 ) , omega-arylalkanoic acids ( 9a-c ) , and the lignan guaiacin ( 10 ) from the seed extract . Compounds 3-5 inhibited P22079 by 86 % , 77 % , and 88 % at 10 ppm , respectively , and compounds 6 and 9a-c showed inhibition at 76 % and 78 % at 100 ppm , respectively . However , compounds 7 and 8 were inactive and lignan 10 exhibited P22079 inhibitory activity by 99 % at 100 ppm compared to commercial antioxidants butylated hydroxyanisole ( BHA ) , butylated hydroxytoluene ( BHT ) , and vitamin E . The flavones 1-5 also inhibited P23219 and -2 enzymes by 50-65 % at 100 ppm . DB04088 showed high but nonselective inhibition of P23219 and P35354 enzymes , when compared to aspirin and DB00482 , a nonsteroidal anti-inflammatory drug ( NSAID ) . Compounds 7 and 10 inhibited P23219 by 60 % and 65 % and P35354 by 37 % and 18 % , respectively , whereas compounds 8 and 9a-c showed little or no activity against these enzymes . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . DB00482 offers a small protection from root resorption associated with orthodontic movement . Tooth movement results from alveolar bone resorption/deposition following application of orthodontic forces , and root resorption can be an undesirable complication associated with this process . No treatment for external root resorption is available to date . OBJECTIVE : To determine if P35354 inhibitors like DB00482 are effective in protecting root resorption associated with orthodontic forces . METHODS : A force of 80 grams was applied to the left maxillary first molars of 7-week-old female Wistar rats using nickel titanium closed coil springs attached to the cervical area of the incisors with 0.010 stainless-steel ligature wires . Twenty animals were divided into three experimental groups : one receiving no treatment , the second receiving 25mg/kg , and the third receiving 50 mg/kg of celecoxib ( DB00482 ) in their drinking water . Rats were maintained on a soft diet and euthanized two weeks after initial placement of the force . Paraffin-embedded sections of the right ( control ) and left ( experimental ) maxillae were stained with H & E and the areas of root resorption were examined by counting the number of lacunaes in the roots . RESULTS : No difference in the distance of tooth movement ( 0.5 mm/two weeks ) was seen in all three groups . The rats that received the low dose of DB00482 showed no statistically significant difference in root resorption than that of the rats that received no dose . The rats that received the high dose of DB00482 showed a lower number of lacunaes ( mean = 3.5 ) than that of the control group ( mean 10.2 ; p=0.02 ) . CONCLUSIONS : Administration of DB00482 during the application of orthodontic forces does not interfere with tooth movement and appears to offer some slight protection against root resorption . Identification of endogenous control genes for normalisation of real-time quantitative PCR data in colorectal cancer . BACKGROUND : Gene expression analysis has many applications in cancer diagnosis , prognosis and therapeutic care . Relative quantification is the most widely adopted approach whereby quantification of gene expression is normalised relative to an endogenously expressed control ( EC ) gene . Central to the reliable determination of gene expression is the choice of control gene . The purpose of this study was to evaluate a panel of candidate EC genes from which to identify the most stably expressed gene(s) to normalise RQ-PCR data derived from primary colorectal cancer tissue . RESULTS : The expression of thirteen candidate EC genes : P61769 , P00492 , P04406 , P60709 , P62937 , O43612 , Q9BV35 , Q8N9I9 , P55056 , Q9UHP6 , P60328 , P30926 and P49406 were analysed in a cohort of 64 colorectal tumours and tumour associated normal specimens . P48061 , P07148 , Q02817 and Q53EL6 genes were chosen as target genes against which a comparison of the effect of each EC gene on gene expression could be determined . Data analysis using descriptive statistics , geNorm , NormFinder and qBasePlus indicated significant difference in variances between candidate EC genes . We determined that two genes were required for optimal normalisation and identified P61769 and P62937 as the most stably expressed and reliable EC genes . CONCLUSION : This study identified that the combination of two EC genes ( P61769 and P62937 ) more accurately normalised RQ-PCR data in colorectal tissue . Although these control genes might not be optimal for use in other cancer studies , the approach described herein could serve as a template for the identification of valid ECs in other cancer types . Phospholipase A₂ activities in skin physiology and pathology . Skin inflammatory diseases are most commonly treated with corticosteroids , especially topical preparations , benefitting from high potency and unparalleled formulation flexibility . However , these benefits are limited due to side effects , especially under long-term use . Non-steroidal anti-inflammatory drugs ( NSAIDs ) which block the P36551 pathways have been used as safer alternatives to corticosteroids , and much effort and resources have been invested in developing P36551 inhibitors . However , synthetic NSAIDs are less potent than steroids , have limited formulation flexibility and have their own safety issues , thereby yielding unsatisfactory results , with some high-profile drugs ( e.g. , the P35354 inhibitors Vioxx , DB00482 ) being withdrawn from the market due to safety concerns . The potency and safety challenges of NSAIDs are related to inter-eicosanoid dynamics , pertaining to their pro-versus anti-inflammatory action , homeostatic functions and tissue-specific activities . Instead , the upstream control of phospholipase A2 ( P04054 ) enzymatic activity , which hydrolyzes cell membrane phospholipids to initiate the eicosanoid production , has been considered for inhibiting eicosanoid activation while maintaining the intricate balance needed for their homeostatic functions . Yet , PLA(2) inhibitors have hardly been tested for treating skin inflammatory/allergic conditions . In this article we review the involvement of PLA(2)s in skin physiology and pathology , and discuss the prospect of PLA(2) inhibition for the treatment of dermatological diseases . Dual carbonic anhydrase -- cyclooxygenase-2 inhibitors . Cyclooxygenase is a key enzyme responsible for metabolisation of arachidonic acid into prostaglandins and thromboxane . This enzyme is the target of non steroidal anti-inflammatory drugs ( NSAIDs ) , used against inflammation and pain . The inducible P35354 was associated with inflammatory conditions , whereas the constitutive form ( P23219 ) was responsible for the beneficial effects of the PGs . This observation led to the development of P35354 inhibitors or " coxibs " of which rofecoxib ( Vioxx ) characterized by a methylsulfone moiety and the sulfonamides celecoxib ( DB00482 ) and valdecoxib ( Bextra ) . Initially described as P35354 " selective " inhibitors , recent reports revealed a nanomolar inhibition activity of the sulfonamide P35354 inhibitors for several carbonic anhydrase ( CA ) isoforms , confirmed by X-ray crystal structures for the adducts of celecoxib and valdecoxib with isozyme CA II . This dual activity may help to explain differences in clinical observation between sulfonamide and methylsulfone P35354 inhibitors . Moreover , the inhibition of CA isozymes , critical for the development and invasion of cancer cells , such as CA II , IX and XII , may constitute an important mechanism of antitumor action of such sulfonamide compounds . Are rofecoxib and celecoxib safer NSAIDS ? NSAIDs work by inhibiting the enzyme cyclo-oxygenase ( P36551 ) , responsible for prostaglandin synthesis . This enzyme exists in two isoforms , P23219 and P35354 . Inhibition of P23219 is thought to be the main cause of the gastrointestinal unwanted effects of NSAIDs , whilst inhibition of P35354 results in anti-inflammatory effects . [ symbol : see text ] DB00533 ( Vioxx -- MSD ) and [ symbol : see text ] celecoxib ( DB00482 -- Searle ) have been developed as selective inhibitors of P35354 . DB00533 is licensed for the symptomatic treatment of osteoarthritis , but not for rheumatoid arthritis . The manufacturer claims that " in clinical studies rofecoxib inhibits P35354 but not P23219 " , has " the power of high-dose NSAIDs -- diclofenac and ibuprofen " and " superior GI safety profile compared to conventional NSAIDs " . Celecoxib is licensed for symptom relief in osteoarthritis and rheumatoid arthritis . The manufacturer claims that celecoxib has " comparable efficacy and superior GI tolerability when compared to diclofenac or naproxen " . Here , we review rofecoxib and celecoxib and consider whether they are safer than conventional NSAIDs . A clinical audit of the prescribing of celecoxib and rofecoxib in Australian rural general practice . AIMS : The new cyclooxygenase-2 ( P35354 ) selective inhibitors , celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) , have been widely prescribed since their launch . No reviews currently appear in the literature of prescribing patterns in Australia . This paper describes a self-audit of the clinical use of selective P35354 inhibitor therapy undertaken with rural general practitioners ( GPs ) in Australia . METHODS : A structured audit form was developed and distributed to interested GPs . The form was self-administered and focused on issues about P35354 inhibitors and the types of patients who were receiving them , e.g. indications , patient demographics , risk factors and drug interactions . RESULTS : A total of 627 patients were recruited ( 569 celecoxib and 58 rofecoxib ) . A range of doses was prescribed . Osteoarthritis was the most common indication ( 68.1 % ) . Risk factors known for the nonselective nonsteroidal anti-inflammatory drugs were identified in 65.1 % of patients , with the most common being advanced age , hypertension and previous peptic ulcer disease . Potential drug interactions were common . A variety of reasons for initiation of therapy was identified ; these included perceived increased efficacy , safety and failure of other treatment . CONCLUSIONS : These results show that P35354 inhibitors are being prescribed for patients with multiple risk factors that may place the patient at increased risk of adverse drug reactions to a P35354 inhibitor . The perception of improved safety and efficacy was common and is of concern . Limitations of the study include the reliance on self-reporting . DB00762 , cisplatin/carboplatin , and P35354 inhibition in small-cell lung cancer . Recent findings indicate significant prolongation of survival and time to disease progression with irinotecan ( CPT-11 , Camptosar ) /cisplatin vs etoposide/cisplatin in extensive-stage small-cell lung cancer , and a larger-scale phase III trial has been planned to provide more definitive data on the benefits of the irinotecan/cisplatin combination in this setting . Early-phase studies indicate that the activity of carboplatin ( DB00958 ) in small-cell lung cancer is comparable to that of cisplatin , and that combining irinotecan on a day 1 and 8 schedule with split-dose carboplatin is feasible . Inhibition of the cyclooxygenase-2 ( P35354 ) enzyme , which is active in tumorigenesis , may augment efficacy and reduce toxicity of platinum/irinotecan combinations . A phase II trial has been designed to compare irinotecan/carboplatin and irinotecan/cisplatin combinations with or without the P35354 inhibitor celecoxib ( DB00482 ) in patients with extensive-stage small-cell lung cancer . Results of these trials will help define the roles of platinum/irinotecan combinations and P35354 inhibition in treatment for small-cell lung cancer . Multifocal synchronous mucinous adenocarcinomas arising in congenital pulmonary airway malformation : a case report with molecular study . AIMS : Congenital pulmonary airway malformation ( CPAM ) is a rare developmental anomaly of the lung . Here , we report a case of mucinous adenocarcinoma arising in CPAM . A 23-month-old boy underwent a thoracoscopic lobectomy of the left upper lobe of the lung based on a presumptive diagnosis of asymptomatic CPAM , found in antenatal sonogram . METHODS AND RESULTS : Histologically , the lesion was consistent with CPAM , Stocker type I . In addition , multiple foci ranging from mucinous epithelial hyperplasia to mucinous adenocarcinoma were detected . All lesions shared the same immunoprofile with the expression of cytokeratin ( CK ) 20 , P98088 , and human epidermal growth factor receptor2 ( P04626 ) , but were negative for CK7 , transcription factor 1 ( Q15669 -1 ) , P15941 , Q99626 , P15056 ( VE1 ) and anaplastic lymphoma kinase ( Q9UM73 ) . K- DB01367 point mutation ( G12V ) was also detected in all micro-dissected mucinous lesions but P00533 mutation was not found . All lesions were consistent with mucinous adenocarcinoma . The patient 's clinical course has been uneventful during the 12-months follow-up period . CONCLUSIONS : This interesting case demonstrated that multiple foci in CPAM can synchronously transform into malignancies . P35354 inhibitors and cancer therapeutics : potential roles for inhibitors of P35354 in combination with cytotoxic therapy : reports from a symposium held in conjunction with the Radiation Therapy Oncology Group June 2001 Meeting . Tumor growth and angiogenesis are interdependent . Cyclooxygenase ( P36551 ) catalyzes the synthesis of prostaglandins ( PGs ) from arachidonic acid . Nonsteroidal antiinflammatory drugs ( NSAIDs ) inhibit P36551 -mediated synthesis of PGs . Although P23219 is constitutively expressed in a wide range of tissues , P35354 is cytokine inducible . Enhanced P35354 expression has been attributed a key role in the development of inflammation and related processes observed in pathologically altered disease states . Two specific P35354 inhibitors , namely , rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) , both oral agents and FDA approved , have been shown preclinically and clinically to have efficacy comparable to that of NSAIDs for relief of pain and inflammation in osteoarthritis , with decreased risk of gastrointestinal damage . Significant preclinical evidence strongly supports the potential role for these inhibitors for the treatment of cancer . On June 29 , 2001 , the Radiation Therapy Oncology Group ( www.rtog.org ) , a National Cancer Institute-sponsored cooperative group , held a 1-day symposium focusing on the potential role of inhibitors of P35354 in the treatment of cancer . This issue of the American Journal of Clinical Oncology contains the summary of those presentations . Celecoxib : a novel treatment for lung cancer . Lung cancer is by far the leading cause of cancer-related deaths . Overall survival is poor and has not improved substantially over the last 50 years . Therefore , it is clear that novel and more effective treatments are needed to improve the outcome of therapy . Recent attention has been drawn to the role of cyclooxygenase ( P36551 ) -2 in the pathogenesis of cancer , and it has been considered as an attractive target for therapeutic and chemopreventive strategies in lung cancer patients . Celecoxib ( DB00482 ) , Pfizer ) , a selective P35354 inhibitor and potent anti-inflammatory agent , has been approved for the treatment of osteoarthritis and rheumatoid arthritis . This orally administered agent is generally well tolerated and has almost no gastrointestinal or renal toxicity . Phase II clinical trials suggest that P35354 inhibition by celecoxib would enhance response to cytotoxic chemotherapy or radiation therapy through interference with cellular proliferation and tumor angiogenic processes , promotion of apoptosis and immune surveillance , or other possible mechanisms . Celecoxib has shown promising antitumor efficacy in lung cancer and a large variety of solid tumors that rely on P35354 -related mechanisms for growth and survival . This article reviews the profile of celecoxib and evidence supporting its role in the therapy of lung cancer . Genetic tools to tailor cancer prevention by NSAIDs . It was shown that NSAIDs , such as aspirin or DB00482 , are effective cancer preventive agents when taken regularly . However , the long-term use of NSAIDs , the cyclooxygenase ( P36551 ) inhibitors , may have significant adverse effects - primarily on the gastrointestinal ( inhibiting P23219 ) and cardiovascular ( inhibiting P35354 ) systems . Genetic analysis of enzymes ( including P36551 ) involved in the prostaglandin synthesis should reveal and predict a person 's benefits vs. toxicity resulting from the NSAID treatment . Targeting angiogenic processes by combination rofecoxib and ionizing radiation . Tumor growth and angiogenesis are interdependent . Cyclooxygenase ( P36551 ) catalyzes the synthesis of prostaglandins from arachidonic acid . Nonsteroidal antiinflammatory drugs ( NSAIDs ) inhibit P36551 -mediated synthesis of prostaglandins . P23219 is constitutively expressed in a wide range of tissues , whereas P35354 is cytokine inducible . Enhanced P35354 expression has been attributed a key role in the development of inflammation and related processes observed in pathologically altered disease states . Two specific P35354 inhibitors , namely rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) , both oral agents and U.S . Food and Drug Administration approved , have been shown preclinically and clinically to have efficacy comparable to that of NSAIDs for relief of pain and inflammation in osteoarthritis , with decreased risk of gastrointestinal damage . Little is known about how angiogenesis is affected by the combination of rofecoxib and radiation . We have evaluated the combination of rofecoxib , at various concentrations , and radiation on cytokine-induced angiogenesis in vitro . We have found that rofecoxib inhibited endothelial cell proliferation , migration , and tube formation ( differentiation ) at clinically relevant doses . In combination with radiation , inhibition of endothelial cell function further increased twofold . The combination of rofecoxib and radiation suggests a complementary strategy with clinical ramifications to target angiogenesis-dependent malignancies . Molecular dynamics simulations of arachidonic acid-derived pentadienyl radical intermediate complexes with P23219 and P35354 : insights into oxygenation regio- and stereoselectivity . The two cyclooxygenase enzymes , P23219 and P35354 , are responsible for the committed step in prostaglandin biosynthesis and are the targets of the nonsteroidal antiinflammatory drugs aspirin and ibuprofen and the P35354 selective inhibitors , DB00482 , Vioxx , and Bextra . The enzymes are remarkable in that they catalyze two dioxygenations and two cyclizations of the native substrate , arachidonic acid , with near absolute regio- and stereoselectivity . Several theories have been advanced to explain the nature of enzymatic control over this series of reactions , including suggestions of steric shielding and oxygen channeling . As proposed here , selective radical trapping and spin localization in the substrate-derived pentadienyl radical intermediate can also be envisioned . Herein we describe the results of explicit , 10 ns molecular dynamics simulations of both P23219 and P35354 with the substrate-derived pentadienyl radical intermediate bound in the active site . The enzymes ' influence on the conformation of the pentadienyl radical was investigated , along with the accessible space above and below the radical plane and the width of several channels to the active site that could function as access routes for molecular oxygen . Additional simulations demonstrated the extent of molecular oxygen mobility within the active site . The results suggest that spin localization is unlikely to play a role in enzymatic control of this reaction . Instead , a combination of oxygen channeling , steric shielding , and selective radical trapping appears to be responsible . This work adds a dynamic perspective to the strong foundation of static structural data available for these enzymes . Interferon-gamma regulation of Clara cell gene expression : in vivo and in vitro . This report demonstrates that Clara cell 10-kDa protein ( P11684 ) mRNA levels are regulated by interferon-gamma ( P01579 ) . An analysis of total lung RNA from mice given P01579 intratracheally showed increased levels of P11684 mRNA compared to control animals but no significant increases in surfactant proteins B and C . These results were confirmed in a Clara cell line , mtCC1-2 , generated from the lungs of a transgenic mouse expressing the SV40 large T antigen under the control of a Clara cell-specific promoter . Significant increases in mtCC1-2 P11684 mRNA levels were observed in a time- and a dose-dependent manner . The expression of transacting factors hepatocyte nuclear factors 3 alpha and 3 beta ( HNF-3 alpha and HNF-3 beta ) were also analyzed , and a transient increase in the expression of HNF-3 beta but not HNF-3 alpha was detected . P24855 footprint analysis identified a signal transducer and activator of transcription ( P35610 ) binding site ( at nucleotides -293 to -284 of P11684 ) adjacent to two thyroid transcription factor-1 ( Q15669 -1 ) binding sites , suggesting a potential interaction between P42224 and Q15669 -1 . This report reinforces the hypothesis that P11684 functions as an anti-inflammatory protein and that increases in P11684 protein may provide additional protection from inflammation and disease in the lung . P35354 inhibitors and DB00482 : safe or suspect ? P35354 inhibitors : better than traditional NSAIDs ? Vioxx and DB00482 may be no less risky than NSAIDs . A real time quantitative PCR analysis and correlation of P23219 and P35354 enzymes in inflamed dental pulps following administration of three different NSAIDs . Dental pain is encountered daily by clinicians . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) commonly used for pain management are traditionally cyclooxygenase-1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) inhibitors , and more recently selective P35354 inhibitors . This study was designed to identify and quantify P23219 and P35354 gene expression level in inflamed rat molar pulps after administration of three NSAIDs : DB00482 , Vioxx , and DB01050 . Fifty male Wistar rats had their first and second molar pulps exposed and sealed with Cavit for 4 days . Rats were randomly divided into the three drug groups and two control groups . RNA was isolated from the rat pulps . Real Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction assay , a relatively new PCR technique , was used to quantify P23219 and P35354 mRNA . Statistical analysis demonstrated no significant differences in P23219 and P35354 levels among the drug groups . However , Vioxx and DB01050 significantly reduced P35354 expression levels compared to inflamed ( positive control ) pulps ( p < 0.05 ) . Inflammation induces mitochondrial dysfunction and dopaminergic neurodegeneration in the nigrostriatal system . Evidence suggests that chronic inflammation , mitochondrial dysfunction , and oxidative stress play significant and perhaps synergistic roles in Parkinson 's disease ( PD ) , where the primary pathology is significant loss of the dopaminergic neurons in the substantia nigra . The use of anti-inflammatory drugs for PD treatment has been proposed , and inhibition of cyclo-oxygenase-2 ( P35354 ) or activation of peroxisome proliferator-activated receptor gamma ( P37231 ) yields neuroprotection in MPTP-induced PD . Lipopolysaccharide ( LPS ) induces inflammation-driven dopaminergic neurodegeneration . We tested the hypothesis that celecoxib ( DB00482 , P35354 inhibitor ) or pioglitazone ( Actos , P37231 agonist ) will reduce the LPS-induced inflammatory response , spare mitochondrial bioenergetics , and improve nigral dopaminergic neuronal survival . Rats were treated with vehicle , celecoxib , or pioglitazone and were intrastriatally injected with LPS . Inflammation , mitochondrial dysfunction , oxidative stress , decreased dopamine , and nigral dopaminergic neuronal loss were observed post-LPS . Celecoxib and pioglitazone provided neuroprotective properties by decreasing inflammation and restoring mitochondrial function . Pioglitazone also attenuated oxidative stress and partially restored striatal dopamine as well as demonstrated dopaminergic neuroprotection and reduced nigral microglial activation . In summary , intrastriatal LPS served as a model for inflammation-induced dopaminergic neurodegeneration , anti-inflammatory drugs provided protective properties , and pioglitazone or celecoxib may have therapeutic potential for the treatment of neuro-inflammation and PD . Current application of selective P35354 inhibitors in cancer prevention and treatment . The multistep process of carcinogenesis , which can take many years , provides many opportunities for intervention to inhibit disease progression . Effective chemoprevention agents may reduce the risk of cancer by inhibiting the initiation stage of carcinoma through induction of apoptosis or DNA repair in cells harboring mutations , or they may act to prevent promotion of tumor growth . Similarly , chemoprevention may entail blocking cancer progression to an invasive phenotype . Over the past decade , in vitro , preclinical , and clinical data have supported the hypothesis that cyclooxygenase ( P36551 ) -2 plays a central role in oncogenesis and that treatment with P35354 inhibitors offers an effective chemoprevention strategy , as exemplified by the activity of celecoxib ( DB00482 ) in familial adenomatous polyposis . These P35354 data have contributed to initiation of clinical trials testing P35354 inhibitors for the chemoprevention of a wide variety of cancers that overexpress P35354 . P35354 inhibitors : do they have a role in emergency department prescribing ? P35354 selective inhibitors ( COXIB or CSI ) have been released with a fanfare as efficacious and safer alternatives to traditional non-steroidal anti-inflammatory drugs . They purport to offer equivalent degrees of analgesia and an improved safety profile . COXIB currently available in Australasia are celecoxib ( DB00482 ) , rofecoxib ( Vioxx ) and etoricoxib ( Arcoxia ) . This review discusses the pharmacology of these agents and reviews recent literature regarding their effectiveness and safety . It endeavours to answer the question ' Should we be using COXIB in emergency departments in Australasia ' ? Rosiglitazone : a disappointing Q9Y2W7 . Dr Steven Nissen is a heart specialist and currently holds the position of chairman of cardiovascular medicine at the Cleveland Clinic , OH , USA . DB00117 work has involved the development of miniaturised ultrasound imaging devices that can be threaded into a patient 's heart that allow measurement of the size and composition of plaques , which indicate early artery damage . The ability to characterize and measure the size of plaques provided a novel , effective method to evaluate the efficacy of anticholesterol medications , and for the past two decades Dr Nissen has been using these and other techniques to examine the efficacy of drugs . He has also developed a strong interest in drug safety . DB00117 work linked P35354 inhibitors such as DB00482 and Vioxx ( Merck , NJ , USA ) with heart attacks , and prevented Merck 's similar product , Arcoxia , from being approved . He also highlighted the serious heart attack risk associated with the experimental drug Pargluva and the drug was subsequently not approved by the US FDA . More recently , Dr Nissen 's work has focused on the drug rosiglitazone , which was shown to have high cardiovascular risks and has since been given a FDA warning . Here , Dr Nissen discusses the publication of the rosiglitazone meta-analysis and why he considers work in this area to be crucially important for patients . Celecoxib-induced upper gastrointestinal hemorrhage and ulceration . P35354 inhibitors are a new class of nonsteroidal anti-inflammatory drugs with a reported benefit of less gastric and duodenal ulceration and hemorrhage . We describe a 67-year-old man taking a higher than usual dose of celecoxib ( DB00482 ) for osteoarthritis with resultant gastric erosions , ulceration , and a significant gastrointestinal ( GI ) hemorrhage . Preventive but not curative efficacy of celecoxib on bladder carcinogenesis in a rat model . To evaluate the effect of a cyclooxygenase 2 inhibitor , celecoxib ( P19835 ) , on bladder cancer inhibition in a rat model , when used as preventive versus as curative treatment . The study comprised 52 male Wistar rats , divided in 5 groups , during a 20-week protocol : control : vehicle , carcinogen : 0.05 % of N-butyl-N-(4-hydroxybutyl) nitrosamine ( BBN ) , P19835 : 10 mg/kg/day of the selective P35354 inhibitor DB00482 , preventive P19835 ( P19835 +BBN-P ) , and curative P19835 ( BBN+ P19835 -C ) groups . Although tumor growth was markedly inhibited by the preventive application of P19835 , it was even aggravated by the curative treatment . The incidence of gross bladder carcinoma was : control 0/8 ( 0 % ) , BBN 13/20 ( 65 % ) , P19835 0/8 ( 0 % ) , P19835 +BBN-P 1/8 ( 12.5 % ) , and BBN+ P19835 -C 6/8 ( 75 % ) . The number and volume of carcinomas were significantly lower in the P19835 +BBN-P versus BBN , accompanied by an ample reduction in hyperplasia , dysplasia , and papillary tumors as well as P35354 immunostaining . In spite of the reduction of tumor volumes in the curative BBN+ P19835 -C group , tumor malignancy was augmented . An anti-inflammatory and antioxidant profile was encountered only in the group under preventive treatment . In conclusion , preventive , but not curative , celecoxib treatment promoted a striking inhibitory effect on bladder cancer development , reinforcing the potential role of chemopreventive strategies based on cyclooxygenase 2 inhibition . Response of chondrocytes to shear stress : antagonistic effects of the binding partners O00206 and caveolin-1 . Osteoarthritis ( OA ) is often a consequence of excessive mechanical loading of cartilage , which produces hydrostatic stress , tensile strain , and fluid flow . Application of high fluid shear to chondrocytes recapitulates the earmarks of OA , as evidenced by the induction of cyclooxygenase-2 , prostaglandins ( PGs ) , and interleukin-6 ( P05231 ) . Here , we delineated the signaling pathway by which high fluid shear mediates the temporal regulation of P05231 synthesis in human chondrocytes . We determined that O00206 ( O00206 ) and caveolin-1 are binding partners in chondrocytes . Their expression is temporally regulated by fluid shear via the sequential up-regulation of microsomal PGE synthase-1 ( mPGES-1 ) and L-PGDS . High shear stress rapidly induces an 8-fold up-regulation of O00206 expression via an mPGES-1-dependent pathway , whereas prolonged shear exposure concurrently down-regulates O00206 by > 4-fold and up-regulates caveolin-1 expression by > 2.5-fold in an L-PGDS-dependent manner . O00206 and caveolin-1 exert opposing effects on the activation of P27361 /2 , P19957 -K and PKA signaling pathways , which , in turn , regulate the NF-κB-dependent P05231 synthesis in a time-dependent fashion . Reconstructing the signaling network regulating shear-induced P05231 expression in chondrocytes may provide insights for developing therapeutic strategies to combat osteoarthritis . DB00945 antagonizes the cytotoxic effect of methotrexate in lung cancer cells . DB00563 ( MTX ) has been widely used for the treatment of cancer and rheumatoid arthritis ( RA ) . DB00945 ( ASA ) is a non-selective cyclooxygenase ( P36551 ) inhibitor that contributes to the treatment of inflammatory conditions such as RA . It has been observed that the antitumor effect of ASA can be attributed to inhibition of cell cycle progression , induction of apoptosis and inhibition of angiogenesis . In the present study , we revealed that the treatment with a combination of MTX and ASA resulted in antagonism of the cytotoxic effect as demonstrated by P50991 and colony formation assays . ASA alleviated the MTX-mediated S phase accumulation and recovered the P55008 phase . MTX-mediated accumulation of the S phase marker cyclin A was also alleviated by ASA . Notably , FAS protein levels were upregulated by MTX in A549 cells . The antagonism of MTX efficacy caused by ASA was accompanied by altered expression of caspase-3 , Bcl-2 and FAS but not dihydrofolate reductase ( P00374 ) . This suggests that the alteration of caspase-3 , Bcl-2 and FAS was involved in the antagonism between ASA and MTX . Exogenously added folic acid reversed the MTX-mediated P00374 inhibition following either MTX or MTX + ASA treatments . Most importantly , we demonstrated for the first time that the commonly used non-steroidal anti-inflammatory drug for headache ASA and possibly other P23219 /2 inhibitors can produce a strong antagonistic effect on the growth inhibition of lung cancer cells when administered in combination with MTX . The clinical implication of our finding is obvious , i.e. , the clinical efficacy of MTX therapy can be compromised by ASA and their concomitant use should be avoided . [ A pop-up menu linked to a computerized drug prescribing system. Prescribing pattern 's feedback via a simple and quick method ] . It takes time for a GP to acquire sufficient experience of a new drug to be able to prescribe competently . This article describes a project studying the use of computerized records to afford a group of GP 's swift feedback on recently introduced drugs of special interest . In the south-east of Sweden a network of primary health care centers has been created in two neighboring counties . The pharmacies of the region are also taking part . When new drugs of particular interest are introduced , each participating GP will automatically see a pop-up menu , asking questions pertaining to each computer-assisted prescription . In the pharmacies , patients are given a questionnaire regarding their expectations with respect to the drug . In this way it will be possible to provide the individual GP swift feedback from a large number of colleagues and patients concerning the drug 's effectiveness in clinical practice . We have now been studying the P35354 inhibitors rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) . Results show that a pop-up menu used in this way provides the general practitioner quick feed-back on prescribing behavior as well as drug effectiveness in clinical practice . [ The effects of DB00482 on mammary tumorigenesis and aging in P04626 /neu transgenic mice ] . It is well known that cyclooxygenase-2 ( P35354 ) plays an important role in the development of many tumors including breast cancer . Our study was concerned with evaluating the effects of the selective P35354 inhibitor , celecoxib , on mammary tumorigenesis and aging in P04626 /neu transgenic mice ( 24 ) . Celecoxib ( celebrex ) 25 mg/kg was administered 5 times a week from the age of 2 months . Twenty-four intact females were in control . Monitoring kept track of tumor detection time , size , presence of lung metastases , food and water consumption , estral function , body weight and temperature . No significant differences between the two groups were reported as far as life-span , tumor growth rate , size and number of metastases to the lung is concerned . To sum up , celecoxib treatment failed to produce any significant effect on carcinogenesis in P04626 /neu transgenic mice . What 's all the fuss ? Safety concerns about P35354 inhibitors rofecoxib ( Vioxx ) and celecoxib ( DB00482 ) . P35354 independent effects of cyclooxygenase-2 inhibitors on oxidative stress and intracellular glutathione content in normal and malignant human B-cells . We recently reported that inhibition of P35354 ( Cox-2 ) reduced human B-CLL proliferation and survival . Herein , we investigated the mechanisms whereby small molecule Cox-2 selective inhibitors , SC-58125 ( a DB00482 analog ) and CAY10404 blunt survival of human B-cell lymphomas and chronic lymphocytic leukemia B-cells . SC-58125 and OSU03012 ( a DB00482 analog that lacks Cox-2 inhibitory activity ) both decreased intracellular glutathione ( DB00143 ) content in malignant human B-cells , as well as in Cox-2 deficient mouse B-cells . This new finding supports Cox-2 independent effects of SC-58125 . Interestingly , SC-58125 also significantly increased B-cell reactive oxygen species ( ROS ) production , suggesting that ROS are a pathway that reduces malignant cell survival . Addition of DB00143 ethyl ester protected B lymphomas from the increased mitochondrial membrane permeability and reduced survival induced by SC-58125 . Moreover , the SC-58125-mediated DB00143 depletion resulted in elevated steady-state levels of the glutamate cysteine ligase catalytic subunit mRNA and protein . These new findings of increased ROS and diminished DB00143 levels following SC-58125 exposure support novel mechanisms whereby a Cox-2 selective inhibitor reduces malignant B-cell survival . These observations also support the concept that certain Cox-2 selective inhibitors may have therapeutic value in combination with other drugs to kill malignant B lineage cells . Chronic anti-inflammatory drug therapy inhibits gel-forming mucin production in a murine xenograft model of human pseudomyxoma peritonei . BACKGROUND : Intraperitoneal accumulation of mucinous ascites in pseudomyxoma peritonei ( PMP ) promotes an inflammatory/fibrotic reaction that progresses to bowel obstruction and eventual patient demise . Cytokines and inflammation-associated transcription factor binding sites , such as glucocorticoid response elements and P35354 , regulate secretory mucin , specifically Q02817 , production . We hypothesized that anti-inflammatory drugs targeting inflammation-associated pathways may reduce mucin production and subsequent disease morbidity in PMP . METHODS : The effects of dexamethasone and DB00482 were assessed in mucin-secreting human colon cancer LS174T cells in vitro and murine xenograft models of LS174T and human appendiceal PMP in vivo by serial parametric measurements , Q02817 transcripts via real-time RT-PCR , and Q02817 protein expression via immunofluorescence assays . RESULTS : Dexamethasone significantly inhibited basal Q02817 mRNA levels in LS174T cells , inhibited mucinous tumor accumulation in an intraperitoneal PMP xenograft model , and prolonged survival in a subcutaneous LS174T xenograft model . DB00482 significantly inhibited sodium butyrate-stimulated Q02817 mRNA levels in LS174T cells and demonstrated a statistically nonsignificant trend toward reduced mucinous tumor growth and prolonged survival in the xenograft models . Q02817 protein analysis by immunofluorescence demonstrated a dual effect of dexamethasone on mucin production and tumor cell count . CONCLUSIONS : Inflammatory mediators are known to regulate mucin production and may promote overexpression of Q02817 by neoplastic cells with goblet cell phenotype in PMP . Anti-inflammatory drugs , dexamethasone and DB00482 , could inhibit extracellular mucin production in PMP by targeting inflammatory cascades and , therefore , may decrease compressive symptoms , increase the disease-free interval , and reduce the extent or frequency of morbid cytoreductive surgeries . Indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) is a potent modulator of LPS-stimulated macrophage functions . Indirubin is a deep-red bis-indole isomer of indigo blue , both of which are biologically active ingredients in Danggui Longhui Wan , an ancient Chinese herbal tea mixture used to treat neoplasia and chronic inflammation and to enhance detoxification of xenobiotics . Multiple indirubin derivatives have been synthesized and shown to inhibit cyclin-dependent kinases ( CDKs ) and glycogen-synthase kinase ( GSK-3β ) with varying degrees of potency . Several indirubins are also aryl hydrocarbon receptor ( P35869 ) agonists , with P35869 -associated activities covering a wide range of potencies , depending on molecular structure . This study examined the effects of indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) , a novel indirubin with potent P40763 inhibitory properties , on basal and LPS-inducible activities in murine RAW264.7 macrophages . Using a focused commercial qRT-PCR array platform ( SuperArray® ) , the effects of E804 on expression of a suite of genes associated with stress and toxicity were determined . Most genes up-regulated by LPS treatment were suppressed by E804 ; including LPS-induced expression of pro-inflammatory cytokines and receptors , apoptosis control genes , and oxidative stress response genes . Using qRT-PCR as a follow up to the commercial arrays , E804 treatment suppressed LPS-induced P35354 , P35228 , P05231 and P22301 gene expression , though the effects on P35228 and P35354 protein expression were less dramatic . E804 also inhibited LPS-induced secretion of P05231 and P22301 . Functional endpoints , including P35228 and lysozyme enzymatic activity , phagocytosis of fluorescent latex beads , and intracellular killing of bacteria , were also examined , and in each experimental condition E804 suppressed activities . Collectively , these results indicate that E804 is a potent modulator of pro-inflammatory profiles in LPS-treated macrophages . Combined preoperative use of celecoxib and gabapentin in the management of postoperative pain . BACKGROUND : In 2005 we reported a study on the efficacy of the preoperative use of the selective P35354 inhibitor celecoxib ( DB00482 ) for reducing both postoperative pain and opioid requirements in patients undergoing bilateral subpectoral breast augmentation . Our findings showed that patients who received 400 mg of celecoxib 30 min before surgery required significantly less postoperative opioid analgesics compared with those given a placebo . DB00996 ( DB00996 ) is an agent commonly used to control neuropathic pain . Here we describe a prospective study assessing the efficacy of preoperative gabapentin in combination with celecoxib for reducing postoperative pain and opioid requirements in elective subpectoral breast augmentation . METHODS : One hundred eighteen patients were given 1200 mg of gabapentin and 400 mg of celecoxib 30-60 min before surgery . From the day of surgery until postoperative day 5 , patients documented any use of analgesics and recorded their degree of pain . Results were then compared with those of our previous study in which only celecoxib was used . RESULTS : The combination of gabapentin and celecoxib was found to be significantly superior ( p < 0.001 ) in reducing postoperative pain and opioid requirements than celecoxib alone in the management of postoperative pain and opioid requirements . CONCLUSION : To decrease postoperative opioid requirements , we recommend 400 mg of celecoxib and 1200 mg of gabapentin taken 30-60 min before surgery by patients undergoing subpectoral breast augmentation or a comparable plastic surgery procedure . P35354 inhibition attenuates antibody responses against human papillomavirus-like particles . Vaccination to generate protective humoral immunity against infectious disease is becoming increasingly important due to emerging strains of virus , poorly immunogenic vaccines , and the threat of bioterrorism . We demonstrate that cyclooxygenase-2 ( Cox-2 ) is crucial for optimal Ab responses to a model vaccine , human papillomavirus type 16 virus-like particles ( HPV 16 VLPs ) . Cox-2-deficient mice produce 70 % less IgG , 50 % fewer Ab-secreting cells , and 10-fold less neutralizing Ab to HPV 16 VLP vaccination compared with wild-type mice . The reduction in Ab production by Cox-2(-/-) mice was partially due to a decrease in class switching . SC-58125 , a structural analog of the Cox-2-selective inhibitor DB00482 reduced by approximately 70 % human memory B cell differentiation to HPV 16 VLP IgG-secreting cells . The widespread use of nonsteroidal anti-inflammatory drugs and Cox-2-selective inhibitory drugs may therefore reduce vaccine efficacy , especially when vaccines are poorly immunogenic or the target population is poorly responsive to immunization . Anti-angiogenic effects of Hypericin-photodynamic therapy in combination with DB00482 in the treatment of human nasopharyngeal carcinoma . Photodynamic therapy ( PDT ) is being investigated as an alternative treatment modality in cancer treatment . It has been shown to induce tumor hypoxia and upregulation of cyclooxygenase-2 ( P35354 ) and vascular endothelial growth factor ( P15692 ) . The objective of this study was to improve in vivo tumor growth control of nasopharyngeal carcinoma ( NPC ) , treated at a subcurative dosage by using a combination of Hypericin-PDT and P35354 inhibitor , DB00482 ( CX ) . The effect of an initial CX dose at 6- and 24-h post-PDT was investigated simultaneously . It was observed that hypoxic NPC/CNE2 cells upregulate both P35354 and P15692 A genes in vitro . In vivo studies , down-regulation of P35354 and hypoxia inducible factor-1alpha ( HIF-1alpha ) genes at 24-h post-PDT and bulk tumor ablation at 48-h post-PDT was observed . However , 24-28 days later regrowth was observed . In a combination treatment , 1st CX dose at 6-h post-PDT had the highest tumor control in which tumors were < or=0.52 cm(3) ( 64.29 % , P < 0.05 ) . However , the tumors administered with a initial dose of CX at 24-h post-PDT had no tumor control . Co-suppression of P35354 , HIF-1alpha and P15692 A genes were observed in tumors with tumor control . Mice without tumor control that were subjected to therapy had increased human P15692 in serum compared to Hypericin-PDT mice . Thus , suggesting that the time of initial administration of CX post-PDT is an important factor for effective tumor control . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . P35354 inhibitors -- IBC conference . 12-13 April 1999 , Coronado , CA , USA . The introduction of celecoxib ( DB00482 , Figure 1 ; GD Searle and Co ) as the first cyclooxygenase ( P36551 )2 selective inhibitor in the US and the expected introduction of rofecoxib ( Vioxx ; Merck and Co Inc ) as the first P35354 inhibitor with an acute pain indication , has prompted interest in this class of drugs as a possible therapeutic improvement on dual P23219 / P35354 inhibitor NSAIDs , currently on the market . Recognition that the P35354 enzyme may have a broader role than pain and inflammation has led to studies investigating the efficacy of P35354 inhibitors for Alzheimer 's disease ( AD ) , stroke , cardiovascular disease and colon cancer . Speakers at the second annual conference sponsored by IBC , addressed issues ranging from the basic concepts of P35354 specificity versus selectivity , pathways and regulatory factors related to P35354 expression , the principles underlying the possible broad implications of the P35354 mechanisms , as well as summaries of recently completed clinical trials supporting the clinical efficacy and safety of P35354 inhibitors in humans . The timeliness of this meeting is emphasized by the recent approval of rofecoxib by the FDA Arthritis Advisory panel and the initial reports in the media of toxicity attributed to celecoxib . Preclinical and limited clinical data presented suggest possible therapeutic roles for selective P35354 inhibitors in neurodegeneration due to both AD and stroke , the prevention and treatment of colon cancer , prevention of premature labor , as well as pain and inflammation . Docking studies on NSAID/ P35354 isozyme complexes using contact statistics analysis . The selective inhibition of P35354 isozymes should lead to a new generation of NSAIDs with significantly reduced side effects ; e.g. celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . To obtain inhibitors with higher selectivity it has become essential to gain additional insight into the details of the interactions between P36551 isozymes-and NSAIDs . Although X-ray structures of P35354 complexed with a small number of ligands are available , experimental data are missing for two well-known selective P35354 inhibitors ( rofecoxib and nimesulide ) and docking results reported are controversial . We use a combination of a traditional docking procedure with a new computational tool ( Contact Statistics analysis ) that identifies the best orientation among a number of solutions to shed some light on this topic . [ Pharma-clinics. The drug of the month. Celecoxib ( DB00482 ) ] . Celecoxib ( DB00482 , Pharmacia ) is a potent and selective inhibitor of the P35354 isoform of cyclooxygenase which is used as nonsteroidal anti-inflammatory drug ( NSAID ) . Its current indications are osteoarthritis and rheumatoid arthritis . The usual recommended daily dosage of celecoxib is 200 mg ( in one or two intakes per day ) , to be increased up to 400 mg ( two intakes per day ) if necessary . Its clinical efficacy seems to be similar to that of other NSAIDs . However , its safety profile , especially gastro-intestinal tolerance and perhaps renal safety , is much better because of the P35354 selectivity . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Influence of polymers on the bioavailability of microencapsulated celecoxib . Celecoxib , a selective P35354 inhibitor , primarily used in treatment of osteoarthritis , rheumatoid arthritis and acute pain was encapsulated in microparticles composed of various polyesters , polymethacrylates or cellulose derivatives used alone or blended . The influence of polymers on microparticle mean diameter , encapsulation efficiency and in vitro and in vivo celecoxib release was investigated . Microparticles were in the size range 11-37 microm . Encapsulation efficiency was optimal due to poor aqueous solubility of celecoxib . Considering in vitro release , microparticles could be divided into drug delivery systems with fast and slow release profiles . Microparticles prepared with poly-epsilon-caprolactone , Eudragit RS and low viscosity ethylcellulose , together with physical mixture of celecoxib with lactose and DB00482 , were tested in vivo . Relative bioavailability of celecoxib was below 20 % in all cases and was probably the consequence of a slow in vivo release of celecoxib from microparticles or low wettability in the case of DB00482 and physical mixture . P35354 inhibitors : a story of greed , deception and death . In 1999 , drug manufacturers introduced a class of NSAIDs called P35354 inhibitors or coxibs . The drugs were avidly promoted directly to the consumers and became bestsellers from the start . Arthritis sufferers were eager to take medications that eased joint pain with less risk of causing gastrointestinal pain , bleeding and other side-effects . In the year after their introduction , doctors wrote over 100 million prescriptions for celecoxib ( DB00482 ) and rofecoxib ( Vioxx ) . DB00482 is the sixth best-selling drug , with sales of more than US $ 4 billion since its debut in 1999 . Vioxx had sales of US $ 2.6 billion in 2001 . However , the coxibs increase the risk of heart attacks and strokes , and their price , in the USA , is obscene . The manufacturers faced a possibly complicit , toothless and bloodless FDA , and used every maneuvering to fleece the patients . We must now reflect on attitudes that we thought only belong to the tobacco industry . Fortunately , safe and active alternatives exist . Effect of celecoxib on capecitabine-induced hand-foot syndrome and antitumor activity . We hypothesized that hand-foot syndrome is an inflammatory phenomenon mediated by the overexpression of cyclooxygenase 2 ( P35354 ) . Therefore , a specific P35354 inhibitor such as celecoxib ( DB00482 ) could attenuate both the incidence and severity of hand-foot syndrome . We undertook a retrospective study comparing the incidences of hand-foot syndrome in 67 patients with metastatic colorectal cancer who took capecitabine ( DB01101 ) with or without celecoxib . Surprisingly , celecoxib seemed to attenuate capecitabine-induced diarrhea as well . DB01101 /celecoxib was also associated with increased tumor response , proportion of stable disease ( 62.5 % vs 22.8 % , P = .001 ) , and increase in median time to tumor progression ( 6 vs 3 months , P = .002 ) compared with capecitabine alone , despite the fact that patients on capecitabine/celecoxib had less favorable disease characteristics ( age , performance status , and prior chemotherapies ) . Overexpression of P35354 , implicated in promoting angiogenesis , enhanced tumor invasiveness , evasion of apoptosis , and immune suppression , is a bona fide molecular target for many solid tumors , including colorectal cancer . Combining capecitabine with celecoxib in the treatment of colorectal cancer has strong preclinical rationales . A prospective study is being designed to evaluate capecitabine and celecoxib with or without epidermal growth factor receptor antagonist ZD1839 in the frontline treatment of metastatic colorectal cancer . These regimens under study are orally based and may significantly impact quality of life in the frontline treatment of metastatic colorectal cancer .

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Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . Unfavourable expression of pharmacologic markers in mucinous colorectal cancer . Patients with mucinous colorectal cancer generally have worse prognoses than those with the nonmucinous variety . The reason for this disparity is unclear , but may result from a differential response to adjuvant chemotherapy . We examined known molecular markers for response to common chemotherapy in these two histological subtypes . In all , 21 patients with mucinous and 30 with nonmucinous Dukes C colorectal cancer were reviewed for demographic data and outcome . Total RNA from the tumours and adjacent normal mucosa was isolated and reverse transcribed . Quantitative expression levels of drug pathway genes were determined using TaqMan RT-PCR ( 5-fluorouracil ( DB00544 ) : P04818 , Q12882 , P19971 ; oxaliplatin : P09211 ( glutathione S-transferase pi ) , P07992 and 2 ; irinotecan : P08183 , Q9UNQ0 , P08684 , P22309 , CES2 , P11387 ) . Mucinous tumours significantly overexpressed both P04818 and P09211 relative to nonmucinous tumours and patient-matched normal mucosa . No significant differences in expression of the remaining markers were found . Mean follow-up was 20 months ; 17 patients had recurrent disease . Among patients receiving DB00544 , those with mucinous tumours experienced shorter disease-free survival ( DFS ) than those with nonmucinous tumours ( median DFS 13.8 vs 46.5 months , P=0.053 ) . Mucinous colorectal cancer overexpresses markers of resistance to DB00544 and oxaliplatin . Likewise , DFS may be decreased in patients with mucinous tumours who receive DB00544 . The presence of mucin should be carefully evaluated in developmental trials of new agents for treating colorectal cancer . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Modulation of estrogen production and 17beta-hydroxysteroid dehydrogenase-type 1 , cytochrome P450 aromatase , c-met , and protein kinase Balpha messenger ribonucleic acid content in rat ovarian granulosa cells by hepatocyte growth factor and follicle-stimulating hormone . P14210 ( P14210 ) suppresses DB00094 -dependent estradiol-17beta ( E(2) ) production in ovarian granulosa cells ( GC ) . The mechanisms of action for P14210 in GC are unknown ; however , activation of the P08581 , c- DB00134 , can induce c-Akt/protein kinase B ( P31749 ) -mediated signal transduction in nonovarian cells . Using immature rat GC , the present study investigated the effects of P14210 within the estrogen biosynthetic pathway , concomitant with changes in c- DB00134 and PKBalpha mRNA expression . Granulosa cells were incubated with androstenedione and DB00094 , P14210 , and/or dibutyryl- DB02527 ( Bu(2)- DB02527 ) . Follicle-stimulating hormone and Bu(2)- DB02527 each stimulated estrone ( E(1) ) and E(2) synthesis at 48 h . P14210 suppressed DB00094 -dependent E(2) , but not E(1) , synthesis . Semiquantitative reverse transcription-polymerase chain reaction showed that P14210 impaired DB00094 -supported 17beta-hydroxysteroid dehydrogenase type-1 ( 17beta-HSD ) and cytochrome P450 aromatase ( P450arom ) mRNA levels . P14210 did not reduce E(2) synthesis or 17beta-HSD and P450arom mRNA expression in the presence of Bu(2)- DB02527 at 48 h . The DB00094 and P14210 each down-modulated c- DB00134 mRNA accumulation , whereas Bu(2)- DB02527 increased c- DB00134 mRNA content . Between 0 and 48 h a biphasic change in PKBalpha mRNA content occurred with either DB00094 or P14210 ; however , PKBalpha mRNA accumulation was augmented by P14210 . Collectively , results suggest that P14210 can suppress E(2) production in GC by disrupting DB02527 -dependent 17beta-HSD and P450arom . Changes in c- DB00134 and PKBalpha mRNA content provide a potential link between P14210 signaling and the DB00094 -dependent mechanisms that control the steroidogenic differentiation of GC . Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization . P10451 -induced , integrin-dependent migration of human mammary epithelial cells involves activation of the hepatocyte growth factor receptor ( DB00134 ) . P10451 ( P10451 ) is a secreted glycophosphoprotein which induces migration of mammary carcinoma cells , and has been implicated in the malignancy of breast carcinoma . P14210 ( P14210 ) induces cell migration of several mammary epithelial cell ( Q9NRJ3 ) lines , via activation of its cognate receptor ( DB00134 ) . This study examines the mechanism of P10451 -induced Q9NRJ3 migration , in terms of the cell surface integrins involved and induction of the P14210 / DB00134 pathway . Three different Q9NRJ3 cell lines were used , representing different stages of tumor progression : 21PT , non-tumorigenic ; 21NT , tumorigenic ; non-metastatic ; and MDA-MB-435 , tumorigenic , highly metastatic . Human recombinant P10451 was found to induce the migration of all three lines . P10451 -induced migration of 21PT and 21NT cells was alphavbeta5 and beta1-integrin dependent , and alphavbeta3-independent , while that of MDA-MB-435 cells was alphavbeta3-dependent . P14210 also induced migration of all three cell lines , and a synergistic response was seen to P14210 and P10451 together . The increased migration response to P10451 was found to be associated with an initial increase in DB00134 kinase activity ( within 30 min ) , followed by an increase in DB00134 mRNA and protein expression . P10451 -induced cell migration is thus mediated by different cell surface integrins in Q9NRJ3 lines representing different stages of progression , and involves activation of the P08581 , DB00134 . A novel role for Bruton 's tyrosine kinase in hepatocyte growth factor-mediated immunoregulation of dendritic cells . The limited success of dendritic cell ( DC ) -based immunotherapy in multiple myeloma is partly due to hepatocyte growth factor ( P14210 ) -induced DC dysfunction . From a therapeutic standpoint , it is important to understand the molecular events involved in inhibition of DC activation/maturation by P14210 . Because Bruton 's tyrosine kinase ( Btk ) negatively regulates maturation and immunostimulatory function of DCs , a role for Btk in P14210 -induced inhibition of both murine and human DCs was investigated . We demonstrate that Btk is a novel proximal component of P14210 -induced c-MET ( P08581 ) signaling . Following P14210 treatment , Btk binds to c-MET and becomes activated . Btk activation in turn blocks the NF-κB pathway and subsequent DC activation via the c-Src-PI3K-AKT-mammalian target of rapamycin ( P42345 ) pathway . Notably , Btk activation is necessary for P14210 -induced association of c-Src and PI3K with c-MET . Furthermore , we provide the first evidence that P14210 inhibits DC activation by inducing autocrine interleukin ( IL ) -10 secretion , which requires activation of Btk . Blocking activation of Btk and its downstream the c-Src-PI3K-AKT- P42345 pathway prevents P14210 -induced P22301 secretion by DCs . In addition , neutralization of P22301 secretion from DCs impaired the inhibitory effect of P14210 on DCs . Thus , our study identifies a novel role for Btk in P14210 -induced DC inhibition . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . P14210 suppresses the anticancer effect of irinotecan by decreasing the level of active metabolite in HepG2 cells . In the liver , carboxylesterase ( CES ) converts irinotecan ( CPT-11 ) to its active metabolite SN-38 , which exerts anticancer effects . SN-38 is metabolized to an inactive metabolite SN-38 glucuronide by uridine 5'-diphospho-glucuronosyltransferase 1A1 ( P22309 ) . Therefore , single nucleotide polymorphisms ( SNPs ) of the P22309 gene are responsible for the severe adverse effects associated with the disruption of SN-38 metabolism . However , despite having SNPs of the P22309 gene , many patients metabolize SN-38 sufficiently to avoid severe adverse effects . Among these patients , we found individuals with elevated serum concentrations of hepatocyte growth factor ( P14210 ) . The aim of this study was to evaluate whether P14210 alters the metabolism of CPT-11 , resulting in a reduction in the anticancer effect of CPT11 . The cytotoxicity of CPT-11 and SN-38 was evaluated in HepG2 cells pretreated with P14210 . Furthermore , we explored the level of expression and mechanisms of activity of CES and P22309 . P14210 suppressed the cytotoxicity of CPT-11 by decreasing intracellular SN-38 levels that resulted from a decrease in CES2 and an increase in P22309 . Furthermore , this P14210 -induced suppression was improved by pretreatment with an inhibitor of P08581 c- DB00134 , and the improvement was synergistically potentiated by epidermal growth factor receptor ( P00533 ) inhibitors . Moreover , P14210 induced phosphorylation of signal transducer and activator of transcription 3 and transactivated P00533 . These results suggest that P14210 is a possible causative agent of acquired clinical resistance in chemotherapy with CPT-11 and could be useful as a predictor of clinical resistance . Additional treatment using c- DB00134 and/or P00533 inhibitors could be a novel strategy to overcome resistance . DB08865 for the treatment of patients with advanced non-small cell lung cancer . DB08865 is a potent small-molecule inhibitor of Q9UM73 ( anaplastic lymphoma kinase ; Q9UM73 ) and hepatocyte growth factor receptor ( P08581 , proto-oncogene c- DB00134 ) . A range of tumors , including subsets of non-small cell lung cancer ( NSCLC ) , anaplastic large cell lymphoma and inflammatory myofibroblastic tumors harbor an Q9UM73 rearrangement that leads to oncogenic activation of Q9UM73 . DB08865 has demonstrated preclinical and clinical activity against such malignancies through inhibition of Q9UM73 , and patients harboring Q9UM73 - rearranged NSCLC have demonstrated high response rates and prolonged progression-free survival in phase I and II studies . In August 2011 , crizotinib was approved for the treatment of advanced Q9UM73 -positive NSCLC . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . TGF-beta-treated microglia induce oligodendrocyte precursor cell chemotaxis through the P14210 -c- DB00134 pathway . In acute experimental autoimmune encephalomyelitis ( EAE ) , demyelination is induced by myelin-specific P01730 (+) T lymphocytes and myelin-specific antibodies . Recovery from the disease is initiated by cytokines which suppress T cell expansion and the production of myelin-toxic molecules by macrophages . Th2/3 cell-derived signals may also be involved in central nervous system ( CNS ) repair . Remyelination is thought to be initiated by the recruitment and differentiation of oligodendrocyte precursor cells ( OPC ) in demyelinated CNS lesions . Here , we report that unlike Th1 cytokines ( P01375 , P01579 ) , the Th2/3 cytokine TGF-beta induces primary microglia from C57BL/6 mice to secrete a chemotactic factor for primary OPC . We identified this factor to be the hepatocyte growth factor ( P14210 ) . Our studies show that P01137 -2-3 as well as IFN-beta induce P14210 secretion by microglia and that antibodies to the P08581 c- DB00134 abrogate OPC chemotaxis induced by TGF-beta2-treated microglia . In addition we show spinal cord lesions in EAE induced in SJL/J mice to contain both OPC and P14210 producing macrophages in the recovery phase , but not in the acute stage of disease . Taken these findings , TGF-beta may play a pivotal role in remyelination by inducing microglia to release P14210 which is both a chemotactic and differentiation factor for OPC . Amelioration of scopolamine induced cognitive dysfunction and oxidative stress by Inonotus obliquus - a medicinal mushroom . The present study was aimed to investigate the cognitive enhancing and anti-oxidant activities of Inonotus obliquus ( Chaga ) against scopolamine-induced experimental amnesia . Methanolic extract of Chaga ( Q9NRJ3 ) at 50 and 100 mg kg (-1)doses were administered orally for 7 days to amnesic mice . Learning and memory was assessed by passive avoidance task ( PAT ) and Morris water maze ( MWM ) test . Tacrine ( DB00382 , 10 mg kg ( -1 ) , orally ( p.o ) ) used as a reference drug . To elucidate the mechanism of the cognitive enhancing activity of Q9NRJ3 , the activities of acetylcholinesterase ( P22303 ) , anti-oxidant enzymes , the levels of acetylcholine ( ACh ) and nitrite of mice brain homogenates were evaluated . Q9NRJ3 treatment for 7 days significantly improved the learning and memory as measured by PAT and MWM paradigms . Further , Q9NRJ3 significantly reduced the oxidative-nitritive stress , as evidenced by a decrease in malondialdehyde and nitrite levels and restored the glutathione and superoxide dismutase levels in a dose dependent manner . In addition , Q9NRJ3 treatment significantly decreased the P22303 activity in both the salt and detergent-soluble fraction of brain homogenates . Further , treatment with Q9NRJ3 restored the levels of ACh as did DB00382 . Thus , the significant cognitive enhancement observed in mice after Q9NRJ3 administration is closely related to higher brain anti-oxidant properties and inhibition of P22303 activity . These findings stress the critical impact of Chaga , a medicinal mushroom , on the higher brain functions like learning and memory . Kringle 1-4 of hepatocyte growth factor inhibits proliferation and migration of human microvascular endothelial cells . P24001 composed of the N-terminal hairpin and subsequent four-kringle domains of hepatocyte growth factor ( P14210 ) is bifunctional , acting as a competitive antagonist for P14210 and an angiogenesis inhibitor . In this study , we determined whether or not four-kringle domains of P14210 ( P04264 -4 ) have anti-angiogenic activity . For this purpose , we prepared recombinant P04264 -4 and P24001 , using the baculovirus expression system . Although P24001 antagonized P14210 -induced DNA synthesis of rat hepatocytes , cell scattering of MDCK cells and the c- DB00134 / P08581 tyrosine phosphorylation in endothelial cells , P04264 -4 failed to antagonize P14210 -induced DNA synthesis , cell scattering and the c- DB00134 / P08581 tyrosine phosphorylation in endothelial cells , thus , indicating that P04264 -4 lacks P14210 -antagonist activity . However , endothelial proliferation and migration induced by P14210 was inhibited by P04264 -4 , similar to the case seen with P24001 . Furthermore , P04264 -4 inhibited the proliferation and migration of human dermal microvascular endothelial cells induced by vascular endothelial growth factor or by basic fibroblast growth factor . We propose that kringle 1-4 of P14210 inhibits angiogenic responses in endothelial cells , independently of P14210 -c- DB00134 signaling pathways . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract .

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Different cholinesterase inhibitor effects on P04141 cholinesterases in Alzheimer patients . BACKGROUND : The current study aimed to compare the effects of different cholinesterase inhibitors on acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) activities and protein levels , in the cerebrospinal fluid ( P04141 ) of Alzheimer disease ( AD ) patients . METHODS AND FINDINGS : AD patients aged 50-85 years were randomized to open-label treatment with oral rivastigmine , donepezil or galantamine for 13 weeks . P22303 and BuChE activities were assayed by Ellman 's colorimetric method . Protein levels were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Primary analyses were based on the Completer population ( randomized patients who completed Week 13 assessments ) . 63 patients were randomized to treatment . DB00989 was associated with decreased P22303 activity by 42.6 % and decreased P22303 protein levels by 9.3 % , and decreased BuChE activity by 45.6 % and decreased BuChE protein levels by 21.8 % . DB00674 decreased P22303 activity by 2.1 % and BuChE activity by 0.5 % , but increased P22303 protein levels by 51.2 % and BuChE protein levels by 10.5 % . Donepezil increased P22303 and BuChE activities by 11.8 % and 2.8 % , respectively . Donepezil caused a 215.2 % increase in P22303 and 0.4 % increase in BuChE protein levels . Changes in mean P22303 -Readthrough/Synaptic ratios , which might reflect underlying neurodegenerative processes , were 1.4 , 0.6 , and 0.4 for rivastigmine , donepezil and galantamine , respectively . CONCLUSION : The findings suggest pharmacologically-induced differences between rivastigmine , donepezil and galantamine . DB00989 provides sustained inhibition of P22303 and BuChE , while donepezil and galantamine do not inhibit BuChE and are associated with increases in P04141 P22303 protein levels . The clinical implications require evaluation . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . P06401 activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells . We previously showed that progesterone ( P4 ) inhibited the proliferation of human umbilical vein endothelial cells ( HUVECs ) through a p53-dependent pathway . Now we investigated further the molecular mechanism underlying the hormone activity . In cultured HUVECs , P4 increased the protein levels of phosphorylated Src ( p-Src ) , P04049 , and P29323 . The levels of p-Src and p-Src-progesterone receptor complex in HUVECs were increased by P4 treatment . These effects were blocked by pretreatment with a progesterone receptor antagonist , DB00834 . The P4-induced increase in p53 transactivity was abolished by pretreatment with Src kinase inhibitors . Moreover , administration with cSrc antisense oligonucleotide prevented the P4-induced increases of the levels of p53 mRNA and protein . These data suggest that P4-induced up-regulation of p53 might be mediated through activation of cSrc . Pretreatment with Src kinase inhibitors also prevented P4-induced membrane translocation of Kras and increases of the protein levels of phosphorylated Raf and phosphorylated P29323 . Transfection with dominant-negative P28482 prevented the P4-induced increases of protein level and promoter activity of p53 and a decrease of thymidine incorporation . P4 also increased nuclear factor-κB ( NF-κB ) nuclear translocation and NF-κB binding onto the p53 promoter . These effects were abolished by pretreatment with P29323 inhibitors . The P4-induced up-regulation of the p53 promoter activity was prevented by preadministration with dominant-negative P28482 or NF-κB inhibitors . Taken together , our data suggest that the cSrc/Kras/ P04049 / P28482 /NF-κB signaling pathway contributes to the P4-induced up-regulation of p53 in HUVECs . These findings highlight progesterone receptor activation of extranuclear signaling pathways in regulating p53 and cell cycle progression in HUVECs . Granulocyte macrophage-colony stimulating factor increases the expression of histamine and histamine receptors in monocytes/macrophages in relation to arteriosclerosis . OBJECTIVE : To study the effect of granulocyte macrophage-colony-stimulating factor ( GM- P04141 ) on histamine metabolism in arteriosclerosis , the expression of histidine decarboxylase ( HDC ; histamine-producing enzyme ) , histamine receptors 1 and 2 ( P35367 and P25021 ) , and GM- P04141 was investigated in human and mouse arteriosclerotic carotid arteries . Furthermore , the molecular mechanisms of GM- P04141 -induced HDC and P35367 expression in monocytic U937 cells were investigated . METHODS AND RESULTS : Immunohistochemistry showed that atherosclerotic human coronary and mouse ligated carotid arteries contained HDC-expressing macrophages . Gene expression of HDC , P35367 , P25021 , and GM- P04141 was also detected in the lesions . In U937 cells , GM- P04141 enhanced histamine secretion and gene expression of HDC and P35367 . A promoter assay showed that GM- P04141 enhanced gene transcription of HDC and P35367 but not P25021 . CONCLUSIONS : The present results indicate that HDC and HHR are expressed in arteriosclerotic lesion , and that GM- P04141 induces HDC and P35367 expression in monocytes . Locally produced histamine might participate in atherogenesis by affecting the expression of atherosclerosis-related genes in monocytes and smooth muscle cells . The presence of histamine-producing macrophages and gene expression of histamine receptors and GM- P04141 was demonstrated in arteriosclerotic lesions . In monocytic U937 cells , GM- P04141 upregulated the expression of histamine and P35367 . Coordinated expression of histamine and its receptors by GM- P04141 would participate in atherogenesis by affecting monocytic and SMC gene expression . Regulation of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor expression by oncostatin M . Oncostatin M ( OM ) is structurally and functionally related to a subclass of hematopoietic cytokines including leukemia-inhibitory factor ( P15018 ) , ciliary neurotrophic factor ( P26441 ) , granulocyte colony-stimulating factor ( DB00099 ) , and interleukin-6 ( P05231 ) . Using human endothelial cells ( O14777 ) as a model for cytokine regulation of hematopoietic growth factor expression , we tested OM as an inducer of colony-stimulating activity . Colony-forming cell assays supplemented with culture supernatants from OM-treated O14777 contained a threefold increase in colony-forming unit granulocyte-macrophage colonies . Specific immunoassay ( enzyme-linked immunosorbent assay ) of culture supernatants indicated that OM treatment of O14777 resulted in a dose- and time-dependent increase in the accumulation of G- P04141 and granulocyte-macrophage P04141 ( GM- P04141 ) ( > 28-fold ) . The ED50 for OM induction of G- P04141 and GM- P04141 protein expression was 17 and 7 pmol/L , respectively . Increased protein expression was associated with a similar increase in steady-state expression of G- P04141 and GM- P04141 mRNA . Furthermore , a period of 12 to 24 hours elapsed before there were measurable increases in P04141 expression , suggesting that OM may stimulate P04141 production through a mechanism requiring the synthesis or activation of a secondary mediating factor or pathway . These findings provide the first evidence that OM may regulate myelopoiesis by inducing the cellular expression of hematopoietic growth factors . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Distinct signalling pathways of murine histamine H1- and H4-receptors expressed at comparable levels in HEK293 cells . DB11320 ( HA ) is recognized by its target cells via four G-protein-coupled receptors , referred to as histamine H1-receptor ( P35367 ) , P25021 , Q9Y5N1 , and Q9H3N8 . Both P35367 and Q9H3N8 exert pro-inflammatory functions . However , their signal transduction pathways have never been analyzed in a directly comparable manner side by side . Moreover , the analysis of pharmacological properties of the murine orthologs , representing the main targets of pre-clinical research , is very important . Therefore , we engineered recombinant HEK293 cells expressing either mouse (m) P35367 or mH4R at similar levels and analyzed HA-induced signalling in these cells . HA induced intracellular calcium mobilization via both mH1R and mH4R , with the mH1R being much more effective . Whereas DB02527 accumulation was potentiated via the mH1R , it was reduced via the mH4R . The regulation of both second messengers via the Q9H3N8 , but not the P35367 , was sensitive to pertussis toxin ( PTX ) . The mitogen-activated protein kinases ( MAPKs ) P29323 1/2 were massively activated downstream of both receptors and demonstrated a functional involvement in HA-induced P18146 gene expression . The p38 MAPK was moderately activated via both receptors as well , but was functionally involved in HA-induced P18146 gene expression only in Q9H3N8 -expressing cells . Surprisingly , in this system p38 MAPK activity reduced the HA-induced gene expression . In summary , using this system which allows a direct comparison of mH1R- and mH4R-induced signalling , qualitative and quantitative differences on the levels of second messenger generation and also in terms of p38 MAPK function became evident . 5-Hydroxytryptamine induces cyclooxygenase-2 in rat vascular smooth muscle cells : Mechanisms involving Src , PKC and MAPK activation [ corrected ] . Considering the importance of 5-hydroxytryptamine ( 5-HT ) and cyclooxygenase ( P36551 ) products in vascular pathology , we investigated the effects of 5-HT on P36551 expression in rat vascular smooth muscle cells ( VSMCs ) , and to provide mechanistic insights into these effects . VSMCs were enzymatically isolated from aortic media of Wistar rats . Incubation of VSMCs with 5-HT for 24h stimulated prostaglandin I(2) production , but this stimulation was completely suppressed by NS-398 , a selective P35354 inhibitor . 5-HT induced transient P35354 , but not P23219 , protein and mRNA expression in concentration- and time-dependent manners . This effect of 5-HT was completely inhibited by sarpogrelate , a 5-HT(2A) receptor antagonist . 5-HT-induced P35354 expression was markedly blunted by Ca(2+) depletion ; GF 109203X , a protein kinase C ( PKC ) inhibitor ; Q99463 , an inhibitor of Src-family tyrosine kinase ( Src ) ; PD 98059 , an inhibitor of extracellular signal-regulated kinase ( P29323 ) activation ; SB 203580 , an inhibitor of p38 mitogen-activated protein kinase ( MAPK ) ; and SP 600125 , an inhibitor of c-Jun N-terminal kinase ( JNK ) . 5-HT activated P29323 and p38 MAPK , followed by JNK activation . Q99463 inhibited these activations , while GF 109203X inhibited only JNK activation . Furthermore , PD 98059 inhibited JNK activation . These results suggest that 5-HT induces P35354 expression in rat VSMCs , and that PKC , Src , and MAPK activation are each essential for the full expression of P35354 pathways . DB00674 ameliorates the impairment of recognition memory in mice repeatedly treated with methamphetamine : involvement of allosteric potentiation of nicotinic acetylcholine receptors and dopaminergic- P27361 /2 systems . DB00674 , a drug used to treat Alzheimer 's disease , inhibits acetylcholinesterase ( P22303 ) and allosterically modulates nicotinic acetylcholine receptors ( nAChRs ) resulting in stimulation of catecholamine neurotransmission . In this study , we investigated whether galantamine exerts cognitive-improving effects through the allosteric modulation of nAChRs in an animal model of methamphetamine ( Meth ) psychosis . The mice treated with Meth ( 1 mg/kg.d ) for 7 d showed memory impairment in a novel object recognition test . DB00674 ( 3 mg/kg ) ameliorated the memory impairment , and it increased the extracellular dopamine release in the prefrontal cortex ( P27918 ) of Meth-treated mice . Donepezil , an P22303 inhibitor ( 1 mg/kg ) increased the extracellular ACh release in the P27918 , whereas it had no effect on the memory impairment in Meth-treated mice . The nAChR antagonist , mecamylamine , and dopamine D1 receptor antagonist , P35240 23390 , blocked the ameliorating effect of galantamine on Meth-induced memory impairment , whereas the muscarinic AChR antagonist , scopolamine , had no effect . The effects of galantamine on extracellular dopamine release were also antagonized by mecamylamine . DB00674 attenuated the defect of the novelty-induced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . The ameliorating effect of galantamine on recognition memory in Meth-treated mice was negated by microinjection of an P29323 inhibitor , PD98059 , into the P27918 . These results suggest that the ameliorating effect of galantamine on Meth-induced memory impairment is associated with indirect activation of dopamine D1 receptor- P27361 /2 following augmentation with dopaminergic neurotransmission in the P27918 through the allosteric activation of nAChRs . DB00674 could be a useful therapeutic agent for treating cognitive deficits in schizophrenia/Meth psychosis , as well as Alzheimer 's disease . Cytokine responses of intestinal epithelial-like Caco-2 cells to non-pathogenic and opportunistic pathogenic yeasts in the presence of butyric acid . Candida albicans , Saccharomyces cerevisiae and their cell wall components , zymosan and glucan , have been shown to stimulate interleukin-8 ( P10145 /CXCL-8 ) production by intestinal epithelial cell-like Caco-2 cells pre-cultured with 10 mM butyric acid . We examined in this study whether these yeasts also altered the production of other cytokines and cyclooxygenases ( COXs ) by Caco-2 cells . Culturing Caco-2 cells with 10 mM butyric acid and 15 % FBS for 4 days enhanced the basal levels of mRNA encoding P05231 , P10145 , Q14116 , monocyte chemoattractant protein ( MCP ) -1 , stem cell factor , transforming growth factor ( TGF ) -beta1 , TGF-beta3 , tumor necrosis factor ( P01375 ) -alpha , P23219 , and P35354 , but not of granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) and TGF-beta2 . The inclusion of live S. cerevisiae or C. albicans further enhanced the production of P10145 , but not of the other cytokines and COXs . The non-pathogenic yeasts , C. kefyr , C. utilis , C. versatilis , Kluyveromyces lactis , K. marxianus , Schizosaccharomyces pombe and Zygosaccharomyces rouxii , used for the production of fermented foods and probiotics , and the opportunistic pathogens , C. glabrata , C. krusei , C. parapsilosis and C. tropicalis , isolated from human tissue samples also enhanced P10145 secretion by Caco-2 cells . P04150 antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK-801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories . Somatostatin preserved blood brain barrier against cytokine induced alterations : possible role in multiple sclerosis . Multiple sclerosis ( MS ) is an inflammatory neurological disorder associated with demyelination , impaired blood brain barrier ( BBB ) , axonal damage and neuronal loss . In the present study , we measured somatostatin ( P61278 ) and tumor necrosis factor-α ( P01375 -α ) like immunoreactivity in P04141 samples from MS and non-MS patients . We also examined the role of P61278 in cytokines and lipopolysaccharide ( LPS ) -induced damage to the BBB using human brain endothelial cells in culture . Most of the cerebrospinal fluid ( P04141 ) samples studied from definite MS patients exhibited lower somatostatin ( P61278 ) -like immunoreactivity and higher expression of P01375 -α in comparison to non-MS patients . Treatment of cells with cytokines and LPS blocked P61278 secretion and decreased P61278 expression . Human brain endothelial cells expressed all five somatostatin receptors ( SSTRs ) with increased expression of P30874 and 4 upon treatment with cytokines and LPS . Cytokines and LPS-induced disruption of the tight junction proteins Zonula occludens ( ZO-1 ) organization was restored in presence of P61278 , P30874 or P31391 selective agonists . Furthermore , inflammation induced changes in extracellular signal-regulated kinases ( P27361 /2 and Q13164 ) signaling and altered expression of endothelial and inducible nitric oxide synthase are modulated in presence of P61278 . These data indicate that decreased levels of P61278 contribute to failure of the BBB in MS . Knockdown of cyclophilin A reverses paclitaxel resistance in human endometrial cancer cells via suppression of MAPK kinase pathways . PURPOSE : Paclitaxel resistance remains to be a major obstacle to the chemotherapy of endometrial cancer . Using proteomic-based approach , we used to identify cyclophilin A ( CypA ) as a potential therapeutic target for endometrial cancer . As a natural continuation , this study aimed to reveal the correlation between CypA and paclitaxel resistance and evaluate the possibility of CypA as a therapeutic target for reversal of resistance . METHODS : Two paclitaxel-resistant endometrial cancer cell sublines O14777 -1-B/TAX and AN3CA/TAX were generated , and expressions of CypA , P-gp , MRP-2 and survivin were demonstrated by Western blotting . CypA was knocked down by RNA interference , and the subsequent effects on the alteration of paclitaxel resistance were examined by MTT , flow cytometry and migratory/invasive transwell assays . MAPK kinases activities were examined by Western blotting . RESULTS : CypA knockdown led to significant inhibition of cell proliferation , induction of apoptosis and suppression of migratory/invasive capacity in O14777 -1-B/TAX and AN3CA/TAX cells when exposed to paclitaxel . CypA knockdown led to reductions in total and phosphorylated MAPK kinases , including Akt , P27361 /2 , p38 MAPK and JNK , in O14777 -1-B/TAX cells . Furthermore , pretreatment with MAPK kinase inhibitors exhibited a synergistic effect in combination with CypA knockdown . CONCLUSIONS : These results demonstrated that CypA expression was up-regulated in paclitaxel-resistant cancer cells , and knockdown of CypA could reverse the paclitaxel resistance through , at least partly , suppression of MAPK kinase pathways , presenting a possibility of CypA serving as a therapeutic target to overcome paclitaxel resistance . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Hyperecho-turbo spin-echo sequences at 3T : clinical application in neuroradiology . BACKGROUND AND PURPOSE : Hyperecho-turbo spin-echo ( hyperTSE ) sequences were developed to reduce the specific absorption rate ( SAR ) , especially at high fields such as 3T and above . The purpose of this study was to quantitatively and qualitatively assess the detection of neuroradiologic pathologies by hyperTSE in comparison with standard turbo spin-echo ( TSE180 degrees ) sequences . MATERIALS AND METHODS : TSE180 degrees and hyperTSE images with parameters adapted for equal P24752 contrast were acquired on a 3T whole-body system in 51 patients with 54 cerebral pathologies . Region-of-interest analysis was performed of signal intensities of pathologies , normal white and gray matter , P04141 , and the SD of noise . Signal intensity-to-noise ratios ( SNRs ) and contrast-to-noise ratios ( CNRs ) for healthy tissues and pathologies were determined . A qualitative rating concerning artifacts , lesion conspicuity , and image quality was performed by 2 experienced neuroradiologists . RESULTS : HyperTSE sequences were equivalent to standard TSE180 degrees sequences for the P21554 of pathologies and of the contrast between gray and white matter . The SNR of gray and white matter and P04141 were also the same . The CNRs of the pathologies in hyperTSE and TSE180 degrees images were strongly correlated with each other ( r = 0.93 , P = .001 ) . The visual rating of images revealed no significant differences between hyperTSE and TSE180 degrees . CONCLUSION : HyperTSE sequences proved to be qualitatively and quantitatively equivalent to TSE180 degrees sequences in the detection of high- and low-signal-intensity lesions . They provide equal P21554 of pathologies and of gray minus white matter and reduce the imaging restrictions of conventional TSE180 degrees imposed by SAR limitations at 3T .

MH_train_1079

MH_train_1079

MH_train_1079

interacts_with DB06695?

multiple_choice

Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . P10275 as a therapeutic target . Androgens function as sex hormone primarily via activation of a single androgen receptor ( AR , or P10275 ) . AR is an important therapeutic target for the treatment of diseases such as hypogonadism and prostate cancer . AR ligands of different chemical structures and/or pharmacological properties are widely used for these therapeutic applications , and all of the AR ligands currently available for therapy modulate AR function via direct binding to the ligand-binding pocket ( P18428 ) of the receptor . In the past ten years , our understanding of AR structure and molecular mechanism of action has progressed extensively , which has encouraged the rapid development of newer generation of AR ligands , particularly tissue-selective AR ligands . With improved tissue selectivity , future generations of AR ligands are expected to greatly expand the therapeutic applications of this class of drugs . This review will provide an overview of the common therapeutic applications of currently available AR ligands , and discussion of the major challenges as well as novel therapeutic strategies proposed for future drug development . Amino acid sequence of trocarin , a prothrombin activator from Tropidechis carinatus venom : its structural similarity to coagulation factor Xa . Among snake venom procoagulant proteins , group II prothrombin activators are functionally similar to blood coagulation factor Xa . We have purified and partially characterized the enzymatic properties of trocarin , the group II prothrombin activator from the venom of the Australian elapid , Tropidechis carinatus ( rough-scaled snake ) . P00734 activation by trocarin is enhanced by Ca2+ , phospholipids , and factor Va , similar to that by factor Xa . However , its amidolytic activity on peptide substrate S-2222 is significantly lower . We have determined the complete amino acid sequence of trocarin . It is a 46,515-Dalton glycoprotein highly homologous to factor Xa and shares the same domain architecture . The light chain possesses an N-terminal Gla domain containing 11 gamma-carboxyglutamic acid residues , followed by two epidermal growth factor ( P01133 ) -like domains ; the heavy chain is a serine proteinase . Both chains are likely glycosylated : the light chain at DB00133 52 and the heavy chain at DB00174 45 . Unlike other types of venom procoagulants , trocarin is the first true structural homologue of a coagulation factor . It clots snake plasma and thus may be similar , if not identical , to snake blood coagulation factor Xa . Unlike blood factor Xa , it is expressed in high quantities and in a nonhepatic tissue , making snake venom the richest source of factor Xa-like proteins . It induces cyanosis and death in mice at 1 mg/kg body weight . Thus , trocarin acts as a toxin in venom and a similar , if not identical , protein plays a critical role in hemostasis . Metalloproteinases control brain inflammation induced by pertussis toxin in mice overexpressing the chemokine P13500 in the central nervous system . Inflammatory leukocytes infiltrate the CNS parenchyma in neuroinflammation . This involves cellular migration across various structures associated with the blood-brain barrier : the vascular endothelium , the glia limitans , and the perivascular space between them . Leukocytes accumulate spontaneously in the perivascular space in brains of transgenic ( Tg ) mice that overexpress P13500 under control of a CNS-specific promoter . The Tg mice show no clinical symptoms , even though leukocytes have crossed the endothelial basement membrane . Pertussis toxin ( PTx ) given i.p. induced encephalopathy and weight loss in Tg mice . We used flow cytometry , ultra-small superparamagnetic iron oxide-enhanced magnetic resonance imaging , and immunofluorescent staining to show that encephalopathy involved leukocyte migration across the glia limitans into the brain parenchyma , identifying this as the critical step in inducing clinical symptoms . Metalloproteinase ( MPs ) enzymes are implicated in leukocyte infiltration in neuroinflammation . Unmanipulated Tg mice had elevated expression of tissue inhibitor of metalloproteinase-1 , matrix metalloproteinase ( MMP ) -10 , and -12 mRNA in the brain . PTx further induced expression of tissue inhibitor of metalloproteinase-1 , metalloproteinase disintegrins-12 , P22894 , and -10 in brains of Tg mice . Levels of the microglial-associated MP P51511 were not affected in control or PTx-treated Tg mice . PTx also up-regulated expression of proinflammatory cytokines IL-1beta and P01375 mRNA in Tg CNS . Weight loss and parenchymal infiltration , but not perivascular accumulation , were significantly inhibited by the broad-spectrum MP inhibitor BB-94/ DB03880 . Our finding that MPs mediate PTx-induced parenchymal infiltration to the chemokine-overexpressing CNS has relevance for the pathogenesis of human diseases involving CNS inflammation , such as multiple sclerosis . Recombinant human prothrombin kringles have potent anti-angiogenic activities and inhibit Lewis lung carcinoma tumor growth and metastases . P00734 , a protein involved in blood coagulation , is a plasma glycoprotein composed of the Gla domain , two adjacent kringle domains , and a serine protease domain . Kringles are triple-disulfide-loop folding domains , which are found in several other blood proteins . In this study , we showed that recombinant human prothrombin kringle-1 , -2. and -1-2 ( rk-1 , -2 , -1-2 ) all have potent anti-angiogenic activities , which inhibit Lewis lung carcinoma ( LLC ) tumor growth and metastases . Recombinant human prothrombin kringles were expressed by an E. coli expression system and purified to apparent homogeneity from crude E. coli extracts . Purified rk-1 , -2 , -1-2 migrated with a molecular mass of 14 , 19 , and 31 kDa , respectively , on sodium dodecyl sulfate-polyacrylamide gel electrophoresis ( SDS-PAGE ) under reducing conditions . rk-1 , -2 , -1-2 exhibited potent inhibitory effects on P09038 -stimulated bovine capillary endothelial cell growth with half-maximal concentrations ( ED50 ) of approximately 41 , 55 , and 156 nM , respectively . All of the recombinant human prothrombin kringles also inhibited angiogenesis in the chorioallantoic membrane ( P62158 ) of chick embryos at a dose of 20 microg . Systemic administration of rk-1 , -2 , -1-2 at a dose of 0.5 mg/kg/day suppressed the growth of primary LLC and at dose of 0.5 and 1.0 mg/kg/day inhibited LLC metastases in C57BL6/J mice lungs through their anti-angiogenic effects . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Effects of G- P04141 on systemic inflammation , coagulation and platelet activation in patients with acute myocardial infarction . INTRODUCTION : In the prospective , randomised , double-blind , placebo-controlled Regenerate Vital Myocardium by Vigorous Activation of Bone Marrow Stem Cells ( REVIVAL ) -2 trial patients with acute myocardial infarction ( AMI ) and successful mechanical reperfusion received granulocyte-colony stimulating factor ( DB00099 , 10 μg/kg KG s.c. ) or placebo for 5 days . Aim of this substudy was to assess the impact of G- P04141 on systemic inflammatory and procoagulant responses and platelet activation . METHODS AND RESULTS : Before and five days after DB00099 ( n=56 ) or placebo ( n=58 ) circulating cytokine concentrations of interleukin ( IL ) -1ß , P05231 , P10145 , P22301 , IL-12 and Tumor-Necrosis Factor-α ( P01375 -α were measured . P00734 fragment F1+2 and Tissue Factor activity served as a measure for activated coagulation . Platelet activation was characterized by cell surface expression of the activated fibrinogen receptor ( O95456 ) , P16109 and P29965 by flow cytometry . Administration of G- P04141 was associated with elevated P01375 -α and CRP concentrations compared to the placebo group after 5 days . Other cytokines ( IL-1ß , P05231 , P10145 , P22301 , IL-12 ) were comparable after treatment with G- P21583 or placebo . Similarly , circulating prothrombin fragments F1+2 , TF activity and platelet activation did not differ in both groups . CONCLUSION : Treatment with G- P04141 in patients with AMI was associated with enhanced proinflammatory P01375 -α and CRP levels but no activation of coagulation . O14788 induces components of the extrinsic coagulation pathway in osteoclasts . P00734 is converted to thrombin by factor Xa in the cell-associated prothrombinase complex . P00734 is present in calcified bone matrix and thrombin exerts effects on osteoblasts as well as on bone resorption by osteoclasts . We investigated whether ( 1 ) osteoclasts display factor Xa-dependent prothrombinase activity and ( 2 ) osteoclasts express critical regulatory components upstream of the prothrombinase complex . The osteoclast differentiation factor O14788 induced formation of multinucleated TRAP positive cells concomitant with induction of prothrombinase activity in cultures of RAW 264.7 cells and bone marrow osteoclast progenitors . Expression analysis of extrinsic coagulation factors revealed that O14788 enhanced protein levels of factor Xa as well as of coagulation factor III ( tissue factor ) . Inhibition assays indicated that factor Xa and tissue factor were involved in the control of prothrombinase activity in O14788 -differentiated osteoclasts , presumably at two stages ( 1 ) conversion of prothrombin to thrombin and ( 2 ) conversion of factor X to factor Xa , respectively . Activation of the extrinsic coagulation pathway during osteoclast differentiation through induction of tissue factor and factor Xa by a O14788 -dependent pathway indicates a novel role for osteoclasts in converting prothrombin to thrombin . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Effect of acetazolamide on aquaporin-1 and fluid flow in cultured choroid plexus . DB00819 ( AZA ) , used in treatment of early or infantile hydrocephalus , is effective in some cases , while its effect on the choroid plexus ( CP ) remains ill-defined . The drug reversibly inhibits aquaporin-4 ( P55087 ) , the most ubiquitous " water pore " in the brain , and perhaps modulation of P29972 ( located apically on CP cells ) by AZA may reduce cerebrospinal fluid ( P04141 ) production . We sought to elucidate the effect of AZA on P29972 and fluid flow in CP cell cultures.CP tissue culture from 10-day Sprague-Dawley rats and a TRCSF-B cell line were grown on Transwell permeable supports and treated with 100 μM AZA . Fluid assays to assess direction and extent of fluid flow , and P29972 expression patterns by immunoblot , Immuncytochemistry ( ICC ) , and quantitative reverse transcriptase polymerase chain reaction ( qRT-PCR ) were performed.Immunoblots and ICC analyses showed a decrease in P29972 protein shortly after AZA treatment ( lowest at 12 h ) , with transient P29972 reduction mediated by mRNA expression ( lowest at 6 h ) . Transwell fluid assays indicated a fluid shift at 2 h , before significant changes in P29972 mRNA or protein levels.Timing of AZA effect on P29972 suggests the drug alters protein transcription , while affecting fluid flow by a concomitant method . It is plausible that other mechanisms account for these phenomena , as the processes may occur independently . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . Inhibition of prothrombin kringle-2-induced inflammation by minocycline protects dopaminergic neurons in the substantia nigra in vivo . P00734 kringle-2 ( pKr-2 ) , a domain of prothrombin , can cause the degeneration of mesencephalic dopaminergic neurons through microglial activation . However , the chemical products that inhibit pKr-2-induced inflammatory activities in the brain are still not well known . The present study investigated whether minocycline , a semisynthetic tetracycline derivative , could inhibit pKr-2-induced microglial activation and prevent the loss of nigral dopaminergic ( DA ) neurons in vivo . To address this question , rats were administered a unilateral injection of pKr-2 in the substantia nigra in the presence or absence of minocycline . Our results show that pKr-2 induces the production of proinflammatory cytokines , such as tumor necrosis factor-α ( P01375 -α ) and interleukin-1β ( IL-1β ) , and inducible nitric oxide synthase from the activated microglia . In parallel , 7 days after pKr-2 injection , tyrosine hydroxylase immunocytochemical analysis and western blot analysis showed a significant loss of nigral DA neurons . This neurotoxicity was antagonized by minocycline and the observed neuroprotective effects were associated with the ability of minocycline to suppress the expression of tumor necrosis factor-α , interleukin-1β , and nitric oxide synthase . These results suggest that minocycline may be promising as a potential therapeutic agent for the prevention of DA neuronal degeneration associated with pKr-2-induced microglial activation . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Proteome of human plasma very low-density lipoprotein and low-density lipoprotein exhibits a link with coagulation and lipid metabolism . Apart from transporting lipids through the body , the human plasma lipoproteins very low-density lipoprotein ( VLDL ) and low-density lipoprotein ( LDL ) are also thought to serve as a modality for intra-organismal protein transfer , shipping proteins with important roles in inflammation and thrombosis from the site of synthesis to effector locations . To better understand the role of VLDL and LDL in the transport of proteins , we applied a combination of LTQ ORBITRAP-XL ( nLC-MS/MS ) with both in-SDS-PAGE gel and in-solution tryptic digestion of pure and defined VLDL and LDL fractions . We identified the presence of 95 VLDL- and 51 LDL-associated proteins including all known apolipoproteins and lipid transport proteins , and intriguingly a set of coagulation proteins , complement system and anti- microbial proteins . P00734 , protein S , fibrinogen γ , P55058 , P11597 , P08571 and P18428 were present on VLDL but not on LDL . Q9UHG3 , dermcidin , cathelicidin antimicrobial peptide , P10646 -1 and fibrinogen α chain were associated with both VLDL and LDL . Apo A-V is only present on VLDL and not on LDL . Collectively , this study provides a wealth of knowledge on the protein constituents of the human plasma lipoprotein system and strongly supports the notion that protein shuttling through this system is involved in the regulation of biological processes . Human diseases related to proteins carried by VLDL and LDL can be divided in three major categories : 1 - dyslipidaemia , 2 - atherosclerosis and vascular disease , and 3 - coagulation disorders . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran .

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interacts_with DB00641?

multiple_choice

The role of de novo ceramide synthesis in the mechanism of action of the tricyclic xanthate D609 . The cytotoxic effects of several chemotherapeutic drugs have been linked to elevated de novo ceramide biosynthesis . However , the relationship between the intracellular site(s) of ceramide accumulation and cytotoxicity is poorly understood . Here we examined the relationship between the site of ceramide deposition and inhibition of protein translation and induction of apoptosis by the antitumor/antiviral xanthate , D609 . In Chinese hamster ovary ( CHO ) - P04264 , P29320 -293 , and NIH-3T3 cells , D609 caused rapid ( 1-5 min ) and sustained eukaryotic initiation factor 2alpha ( eIF2alpha ) phosphorylation followed by apoptosis after 24 h . Concurrently , D609 stimulated de novo ceramide synthesis and increased ceramide mass 2-fold by 2 h in CHO- P04264 cells . In D609-treated CHO- P04264 cells , sphingomyelin synthesis was stimulated by brefeldin A , and P01031 - P28068 -ceramide transport to the Golgi apparatus was blocked , indicating ceramide accumulation in the endoplasmic reticulum ( ER ) . However , D609-mediated eIF2alpha phosphorylation , inhibition of protein synthesis , and apoptosis in CHO- P04264 cells were not attenuated by fumonisin B1 or l-cycloserine . Interestingly , short-chain ceramide promoted eIF2alpha phosphorylation and inhibited protein synthesis in CHO- P04264 cells , indicating that the effectiveness of endogenous ceramide could be limited by access to signaling pathways . Thus , expansion of the ER ceramide pool by D609 was not implicated in early ( eIF2alpha phosphorylation ) or late ( apoptotic ) cytotoxic events . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Determination of MK-0767 enantiomers in human plasma by normal phase LC-MS/MS . A sensitive and selective analytical method for the enantioselective determination of MK-0767 , a dual peroxisome proliferator-activated receptor ( Q07869 ) alpha/gamma agonist , in human plasma has been developed and validated . The chromatography is based on normal-phase chiral separation on a Kromasil , 5 microm , CHI- P28068 250 mm x 4.6 mm column . The detection involves the direct introduction of the normal phase eluent into MS/MS without the addition of a post-column reagent . Atmospheric pressure chemical ionization ( APcI ) mode was selected as the ion source in this method . With proper sample handling and processing procedures , ex vivo interconversion of the enantiomers was kept to minimum during sample collection , preparation and short term storage of frozen human plasma samples . The method was successfully utilized to determine the concentrations of MK-0767 enantiomers in human plasma to support pharmacokinetic investigation in man . P01130 -deficient mice are protected against lethal endotoxemia and severe gram-negative infections . Lipoproteins can bind lipopolysaccharide ( LPS ) and decrease the LPS-stimulated production of pro-inflammatory cytokines . We investigated the effect of increased plasma concentrations of low-density-lipoproteins ( LDL ) on survival and cytokine production after a lethal challenge with either LPS or live Gram-negative bacteria in P01130 deficient mice ( P01130 -/- ) . The P01130 -/- mice challenged with LPS had an eightfold increased LD50 when compared with the wild type controls ( C57Bl/6J ) , while tumor necrosis factor alpha ( TNFalpha ) and interleukin-1 alpha ( P01583 ) plasma concentrations were decreased twofold . P01130 -/- mice had significantly lower and delayed mortality than control mice after infection with Klebsiella pneumoniae . No differences in the outgrowth of bacteria in the organs were present between the two groups , while circulating cytokine concentrations were decreased twofold in P01130 -/- mice . In contrast , the LPS-stimulated in vitro production of cytokines by peritoneal macrophages of P01130 -/- mice was significantly increased compared with controls . This increase was associated with enhanced specific binding of LPS to the macrophages of P01130 -/- mice . In conclusion , endogenous LDL can protect against the lethal effects of endotoxin and Gram-negative infection . At least part of this protection is achieved through decreased in vivo production of pro-inflammatory cytokines , in spite of increased cytokine production capacity . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Genetic influences on sarcoidosis . To investigate the genetic influences underlying the development of sarcoidosis , HLA class II genotyping was performed in Japanese patients with sarcoidosis and healthy controls using the PCR-RFLP method . The frequencies of both DR52 group antigen-associated alleles ( HLA- Q8IUH3 *11 , - Q8IUH3 *12 and - Q8IUH3 *14 ) and Q8IUH3 *08 alleles were higher in the patient group , suggesting that the common , specific amino acid residue on the Q8IUH3 molecule of these alleles may determine susceptibility to sarcoidosis . Alternatively , it is possible that another susceptibility gene , linked to these Q8IUH3 alleles , exists within the MHC region . We screened the P01375 , P01374 , P0DMV8 and Hum70t genes around the class III region , as well as the P28067 and - P28068 genes in the class II region , for genetic polymorphism in sarcoidosis . None of these genes suggested a susceptibility to sarcoidosis . These studies support the thesis that one of the major genetic factors controlling the development of sarcoidosis is located within the Q8IUH3 locus in the HLA class II region . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P10275 -mediated antagonism of estrogen-dependent low density lipoprotein receptor transcription in cultured hepatocytes . Postmenopausal women receiving hormone replacement therapy have a lower risk of coronary heart disease than women who do not receive hormone treatment . Multiple mechanisms are likely to underlie estrogen 's cardioprotective action , including lowering of plasma low density lipoprotein ( LDL ) cholesterol . Using an in vitro system exhibiting normal regulation of P01130 ( P01130 ) gene transcription , we show that 17beta-estradiol activates the P01130 promoter in transiently transfected HepG2 cells . P01130 activation by estrogen in HepG2 cells is dependent on the presence of exogenous estrogen receptor , and the estrogen-responsive region of the P01130 promoter colocalizes with the sterol response element previously identified . The estrogen response is concentration dependent , saturable , and sensitive to antagonism by estrogen receptor antagonists . Further , we show that compounds with androgen receptor agonist activity attenuate the estrogen-induced up-regulation of P01130 in our model system . Progestins with androgen receptor agonist activity , such as medroxyprogesterone acetate , also suppress estrogen 's effects on P01130 expression through their androgenic properties . Characterization of the interplay between these hormone receptors on the P01130 in vitro system may allow a better understanding of the actions of sex steroids on P01130 gene expression and their roles in cardiovascular disease . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . Adhesion molecules and atherogenesis . Atherosclerosis is an inflammatory disease of the vessel wall characterized by monocyte infiltration in response to pro-atherogenic factors such as oxidized lipids . Recently , the role of specific adhesion molecules in this process has been explored . The endothelium overlying atherosclerotic lesions expresses P16109 and the shoulder regions express vascular cell adhesion molecule-1 ( P19320 ) and intercellular adhesion molecule-1 ( P05362 ) , which is also expressed on endothelium in regions not prone to plaque development . Serum levels of soluble P16109 , P05362 and P19320 are elevated in patients with angina pectoris or peripheral atherosclerotic disease . Reconstituted in vitro systems using monocytes on cytokine-activated endothelial cells under shear flow suggested the involvement of P16109 , P14151 , P19320 , its ligand , VLA-4 integrin and P05107 integrins . Studies of monocyte adhesion in isolated perfused carotid arteries harvested from atherosclerotic ( apoE-/- ) mice show a predominant involvement of P16109 and its ligand P16109 glycoprotein-1 ( Q14242 ) in rolling and of VLA-4 and P19320 in firm adhesion . Consistent with these findings , apoE-/- mice that are also deficient for P16109 show significantly reduced atherosclerotic lesion sizes and are almost completely protected from neointimal growth after vascular injury . Milder effects are also seen in the low-density lipoprotein ( LDL ) receptor deficient ( P01130 -/- ) mouse . In a high cholesterol/cholate model , a role of P05362 and P05107 integrins was also shown , but this awaits confirmation in more physiologic models . Transient blockade of the VLA-4/ P19320 adhesion pathway by antibodies or peptides in apoE-/- or P01130 -/- mice reduced monocyte and lipid accumulation in lesions . These data suggest that P16109 , Q14242 , VLA-4 and P19320 are the most important adhesion molecules involved in monocyte recruitment to atherosclerotic lesions . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . An improved method for determining proteoglycans synthesized by chondrocytes in culture . An improved micro method for measuring sulfated glycosaminoglycans ( S-GAG ) in chondrocyte cultures using 1,9-Dimethylmethylene Blue ( P28068 ) has been developed . By increasing the protein concentration in the P28068 assay a soluble GAG- P28068 complex is prolonged . Without bovine serum albumin ( BSA ) in the phosphate-buffered saline ( PBS ) medium , the half time for loss of absorbance was 18 min ; with 1 % BSA-PBS there was no loss of absorbance over this time period . The limit of detection in a 96 well microtiter plate assay was 2 micrograms/ml ; for a cuvette assay it was 1 microgram/ml . Collagen , DNA and RNA did not interfere with this assay . DB08818 caused an increase in absorbance at 530 nm that was lost by preincubating with Streptomyces hyaluronidase . The increase in absorbance was due to a turbidity change because there was no color shift from 600 to 530 nm but rather a uniform increase in absorbance between 400 to 700 nm . To validate the assay , the S-GAG was measured in conditioned medium from primary bovine articular chondrocyte monolayer cultures . A protein synthesis inhibitor , cycloheximide , blocked proteoglycan synthesis by greater than 90 % . A cytokine , P01583 , caused a dose-dependent decrease in proteoglycan accumulation . P34059 ABC digestion of the chondrocyte conditioned medium completely prevented reactivity with the P28068 . By preincubating samples with specific enzymes , different types of S-GAG can be measured with this assay . This assay can be used to measure changes in proteoglycans synthesized by chondrocytes . DB01645 stimulates electrogenic Cl(-) secretion in mouse jejunum . We used the short-circuit current ( I(sc) ) technique to investigate the effects of the isoflavone genistein on the electrogenic Cl(-) secretion of the mouse jejunum . DB01645 stimulated a sustained increase in I(sc) that was dose dependent . DB00887 inhibited 76 +/- 5 % of the genistein-stimulated I(sc) consistent with activation of Cl(-) secretion . DB01645 failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin . In addition , forskolin had a reduced effect on I(sc) of the mouse jejunum in the presence of genistein . DB01016 , a blocker of P13569 , eliminated the genistein-stimulated increase of I(sc) and reduced the forskolin-activated I(sc) . Clotrimazole , a Ca(2+)-activated K(+) channel blocker , failed to reduce the genistein-stimulated I(sc) . Vanadate , a blocker of tyrosine-dependent phosphatases , reduced the genistein-activated I(sc) . Tyrphostin A23 , a tyrosine kinase inhibitor , reduced basal I(sc) , after which genistein failed to stimulate I(sc) . These data suggest that genistein activated a sustained Cl(-) secretory response of the mouse jejunum and that the effect of genistein was via a tyrosine-dependent phosphorylation pathway . UMD ( Universal mutation database ) : a generic software to build and analyze locus-specific databases . The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases . In the near future , the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers , which is growing in frequency in industrial states . The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations . Other fields such as molecular epidemiology will also be developed using these new data . Consequently , the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line . To meet this goal , we devised a generic software called UMD ( Universal Mutation Database ) . It was developed as a generic software to create locus-specific databases ( LSDBs ) with the 4(th) Dimension(R) package from ACI . This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data . Thanks to the flexible structure of the UMD software , it has been successfully adapted to nine genes either involved in cancer ( P25054 , P04637 , P06400 , O00255 , Q09428 , P40337 , and P19544 ) or in genetic diseases ( P35555 and P01130 ) . Four new LSDBs are under construction ( P49748 , P11310 , KIR6 , and P29400 ) . Finally , the data can be transferred to core databases . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . Plasmodium falciparum exports the Golgi marker sphingomyelin synthase into a tubovesicular network in the cytoplasm of mature erythrocytes . This work describes two unusual features of membrane development in a eukaryotic cell . ( a ) The induction of an extensive network of tubovesicular membranes by the malaria parasite Plasmodium falciparum in the cytoplasm of the mature erythrocyte , and its visualization with two ceramide analogues P01031 - P28068 -ceramide and P13671 -NBD-ceramide . " Sectioning " of the infected erythrocytes using laser confocal microscopy has allowed the reconstruction of detailed three-dimensional images of this novel membrane network . ( b ) The stage-specific export of sphingomyelin synthase , a biosynthetic activity concentrated in the Golgi of mammalian cells , to this tubovesicular network . Evidence is presented that in the extracellular merozoite stage the parasite retains sphingomyelin synthase within its plasma membrane . However , intracellular ring- and trophozoite-stage parasites export a substantial fraction ( approximately 26 % ) of sphingomyelin synthase activity to membranes beyond their plasma membrane . Importantly we do not observe synthesis of new enzyme during these intracellular stages . Taken together these results strongly suggest that the export of this classic Golgi enzyme is developmentally regulated in Plasmodium . We discuss the significance of this export and the tubovesicular network with respect to membrane development and function in the erythrocyte cytosol . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation . Activity , pharmacological inhibition and biological regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Trypanosoma brucei . Activity of hydroxymethylglutaryl-coenzyme A ( HMG- DB01992 ) reductase , the key enzyme in the biosynthesis of steroids and polyisoprenoids in mammalian cells , has been detected in both the bloodstream form and the culture-adapted procyclic form of Trypanosoma brucei ( 3.7 +/- 0.6 and 12.7 +/- 1.8 pmol mevalonate produced min-1 ( mg cell protein ) -1 , respectively ) . The enzyme activity is enriched 6-fold in microsomal fractions . Several competitive inhibitors of mammalian P04035 , including synvinolin ( simvastatin ) , inhibit the multiplication of both forms of trypanosome in vitro ( IC50 , approx. 25-50 microM after 2-3 days ) . This growth inhibition is potentiated by agents interfering with the exogenous supply of cholesterol , such as antibodies blocking the low-density lipoprotein ( LDL ) receptor , or 5 microM chloroquine . Conversely , growth inhibition by synvinolin can be largely reverted either by 300 nM LDL or by products of the mevalonate pathway , such as 20 mM mevalonate and in procyclics by 100 microM squalene or cholesterol . In procyclics , low concentrations of synvinolin selectively inhibit the incorporation of [14C]acetate into sterols , but not into fatty acids . These results argue for a critical role in trypanosomes of a mevalonate pathway , that is involved in the biosynthesis of sterol and probably of other metabolites . The P04035 activity is decreased 2-fold in procyclics incubated with 4 mM mevalonate and increased 2-fold in the presence of 2.5 microM synvinolin . DB00641 also upregulates LDL binding up to 4-fold . These data suggest that P04035 and P01130 expression are regulated in T. brucei as in mammalian cells , to ensure sterol homeostasis . Functional alterations in endothelial NO , PGI₂ and EDHF pathways in aorta in ApoE/ P01130 -/- mice . Adequate endothelial production of nitric oxide ( NO ) , endothelium-derived hyperpolarizing factor ( EDHF ) , and prostacyclin ( PGI₂ ) is critical to the maintenance of vascular homeostasis . However , it is not clear whether alterations in each of these vasodilatory pathways contribute to the impaired endothelial function in murine atherosclerosis . In the present study , we analyze the alterations in NO- , EDHF- and PGI₂-dependent endothelial function in the thoracic aorta in relation to the development of atherosclerotic plaques in apoE/LDLR⁻/⁻ mice . We found that in the aorta of 2-month-old apoE/LDLR⁻/⁻ mice there was no lipid deposition , subendothelial macrophage accumulation ; and matrix metalloproteinase ( MMP ) activity was low , consistent with the absence of atherosclerotic plaques . Interestingly , at this stage the endothelium was already activated and hypertrophic as evidenced by electron microscopy , while acetylcholine-induced NO-dependent relaxation in the thoracic aorta was impaired , with concomitant upregulation of cyclooxygenase-2 ( P35354 ) /PGI₂ and EDHF ( epoxyeicosatrienoic acids , EETs ) pathways . In the aorta of 3-6-month-old apoE/LDLR⁻/⁻ mice , lipid deposition , macrophage accumulation and MMP activity in the intima were gradually increased , while impairment of NO-dependent function and compensatory upregulation of P35354 /PGI₂ and EDHF pathways were more accentuated . These results suggest that impairment of NO-dependent relaxation precedes the development of atherosclerosis in the aorta and early upregulation of P35354 /PGI₂ and EDHF pathways may compensate for the loss of the biological activity of NO . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Products of the unc-52 gene in Caenorhabditis elegans are homologous to the core protein of the mammalian basement membrane heparan sulfate proteoglycan . Mutations in the unc-52 gene of Caenorhabditis elegans affect attachment of the myofilament lattice to the muscle cell membrane . Here , we demonstrate that the unc-52 gene encodes a nematode homolog of perlecan , the mammalian basement membrane heparan sulfate proteoglycan . The longest potential open reading frame of this gene encodes a 2482-amino-acid protein with a signal peptide and four domains . The first domain is unique to the unc-52 polypeptide , whereas the three remaining domains contain sequences found in the P01130 ( domain II ) laminin ( domain III ) and N- P62158 ( domain IV ) . We have identified three alternatively spliced transcripts that encode different carboxy-terminal sequences . The two larger transcripts encode proteins containing all or part of domain IV , whereas the smaller transcript encodes a shortened polypeptide that completely lacks domain IV . We have determined that the disorganized muscle phenotype observed in unc-52(st196) animals is caused by the insertion of a Tc1 transposon into domain IV . Two monoclonal antibodies that recognize an extracellular component of all contractile tissues in C. elegans fail to stain embryos homozygous for a lethal unc-52 allele . We have mapped the epitopes recognized by both monoclonal antibodies to a region of domain IV in the unc-52-encoded protein sequence . Determination of free N-acetylneuraminic acid in human body fluids by high-performance liquid chromatography with fluorimetric detection . Determinations of both the free and bound form of N-acetyl-neuraminic acid ( NANA ) in several human body fluids , such as serum , cerebrospinal fluid ( P04141 ) , saliva , urine , amniotic fluid , and milk were carried out by HPLC with fluorimetric detection . The method utilized 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride ( P28068 ) as a fluorimetric derivatizing reagent . Free-form NANA was obtained from the body fluids after ultrafiltration with Microcon 10 ( YM-10 cellulose membrane , filtration limit M(r) = 10,000 , Amicon ) . The P28068 derivative of NANA was separated isocratically by a Nucleosil 5C18 column with a mixture of 0.1 M sodium phosphate buffer ( pH 2.0 ) -methanol ( 75:25 , v/v ) . A gradient elution system was used for urine analysis . Analysis times were 10-30 min . Recoveries of free NANA by ultrafiltration were satisfactory : 95.66 +/- 1.80 % for serum and 97.27 +/- 1.55 % for P04141 , respectively . The high sensitivity and specificity render this method applicable to all the body fluids tested . Although a physiological role for free NANA has not yet been elucidated , the method presented promises to contribute to the basic understanding of the NANA metabolism . Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization . Medial Expression of P01375 -α and P01375 Receptors Precedes the Development of Atherosclerotic Lesions in P02649 /LDL Receptor Double Knockout Mice . P01375 -α is present in atherosclerotic lesions , activates endothelial adhesion molecule expression , stimulates the release of proinflammatory cytokines and matrix metalloproteinases and promotes smooth muscle cell proliferation and migration . Taken together these observations suggest that P01375 -α may be functionally involved in early atherosclerosis development . To further evaluate this hypothesis we compared vascular P01375 -α and P01375 receptor expression in atherosclerosis-susceptible apoE(-/-)/ P01130 (-/-) mice and control C57BL/6 mice . The aortas of 8 week old apoE(-/-)/LDLreceptor(-/-) mice displayed immunoreactivity for P01375 -α as well as P01375 p55 and p75 receptors ( 2.1 ± 1.6 % , 5.6 ± 1.5 % and 3.6 ± 1.3 % of total media area , respectively ) , but did not have any detectable lesions . A marginal increase in P01375 -α and P01375 receptor immunoreactivity was observed at 12 weeks and atherosclerotic plaques were detected in 1 out of 5 animals . At 16 weeks P01375 -α expression in the media was increased more than four-fold as compared with 8 week old mice , and atherosclerosis was widespread . P01375 -α immunoreactivity was also observed in all plaques . In addition , at the same age a tendency towards increased P01375 -α mRNA levels was detected in the double knockout mice compared to age-matched controls . A further increase in P01375 -α and P01375 receptor immunoreactivity as well as plaque size was observed at 20 weeks . With only a few exceptions , no P01375 -α or P01375 receptor immunoreactivity was detected in C57BL/6 control mice . These findings demonstrate that medial P01375 -α and P01375 receptor expression precedes lesion formation in apoE(-/-)/ P01130 (-/-) mice . Metabolic fate of 2,2-dimethylbutyryl moiety of simvastatin in rats : identification of metabolites by gas chromatography/mass spectrometry . Metabolic pathways of simvastatin ( DB00641 ) , a lactone prodrug of an inhibitor of P04035 , were elucidated in male rats , using the [ 14C ] -labelled compound . Evidence has been obtained for hydrolysis of simvastatin and its metabolites at their 2,2-dimethylbutyryl moieties . Metabolites identified in plasma were 2,2-dimethylbutyric acid ( P28068 ) , 2,2-dimethyl-3-hydroxybutyric acid ( DMHB ) and an open chain hydroxy acid of simvastatin : metabolites identified in urine were DMHB , a glucuronide and the glycine conjugate of P28068 . They were characterized by gas chromatography/electron impact and chemical ionization mass spectrometry as phenacyl or pertrimethylsilylated derivatives . The structures of the metabolites and the aglycone of the glucuronide were confirmed as phenacyl esters by comparison of their chromatographic data and mass spectra with those of the phenacyl derivatives of authentic compounds . Molecular genetic studies in monogenic and polygenic human diseases . The main goal of this study was to determine and characterise the types of mutations in two monogenic human disorders : cystic fibrosis ( CF ) and Duchenne/Becker muscular dystrophy ( P11532 , BMD ) and the susceptibility allele frequency in a polygenic disease : type I insulin-dependent diabetes mellitus ( IDDM ) . After analysing 220 chromosomes for mutations in the CF ( Cystic Fibrosis Transmembrane Conductance Regulator = P13569 ) gene , delta F508 mutation was most abundant ( 41 % ) and out of the non-delta F508 CF mutations 5 % was identified as G542X , G551D , R553X , N1303K and W1282X . The CF haplotype analysis by using linked markers to the P13569 gene revealed that the CF " B " haplotype occurred in 66.7 % of patients , and this haplotype was 57.2 % in patients carrying the delta F508 mutation . Prenatal genetic diagnosis for CF was performed in 10 fetuses : 3 were affected , 6 were carriers , and 1 without any CF mutation . Fifty % of 66 patients with P28068 /BMD muscular dystrophy had one or more exon deletions in the dystrophin gene . Eighty-five % of the deletions occurred at the 3' and 15 % at the 5' end of the gene . Out of the three prenatal diagnosis in one case P11532 was substantiated . Thirty-six % of 50 patients with IDDM possessed four , 44 % three and 20 % two susceptibility markers in the HLA-DQA1 , -DQB1 region . The onset of the disease correlated with the number of susceptibility alleles . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress .

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DB00470 -induced neurotoxicity depends on P21554 receptor-mediated c-Jun N-terminal kinase activation in cultured cortical neurons . Delta9- DB00470 ( THC ) , the main psychoactive ingredient of marijuana , induces apoptosis in cultured cortical neurons . THC exerts its apoptotic effects in cortical neurons by binding to the P21554 cannabinoid receptor . The P21554 receptor has been shown to couple to the stress-activated protein kinase , c-Jun N-terminal kinase ( JNK ) . However , the involvement of specific JNK isoforms in the neurotoxic properties of THC remains to be established . The present study involved treatment of rat cultured cortical neurons with THC ( 0.005-50 microM ) , and combinations of THC with the P21554 receptor antagonist , AM 251 ( 10 microM ) and pertussis toxin ( PTX ; 200 ng ml-1 ) . Antisense oligonucleotides ( AS ) were used to deplete neurons of P45983 and P45984 in order to elucidate their respective roles in THC signalling . Here we report that THC induces the activation of JNK via the P21554 receptor and its associated G-protein , Gi/o . Treatment of cultured cortical neurons with THC resulted in a differential timeframe of activation of the P45983 and P45984 isoforms . Use of specific P45983 and P45984 AS identified activation of caspase-3 and DNA fragmentation as downstream consequences of P45983 and P45984 activation . The results from this study demonstrate that activation of the P21554 receptor induces JNK and caspase-3 activation , an increase in Bax expression and DNA fragmentation . The data demonstrate that the activation of both P45983 and P45984 isoforms is central to the THC-induced activation of the apoptotic pathway in cortical neurons . Effects of delta9-THC on P01282 -induced prolactin secretion in anterior pituitary cultures : evidence for the presence of functional cannabinoid P21554 receptors in pituitary cells . Peripheral administration of cannabinoid P21554 receptor agonists to laboratory rats induce a brief rise in plasma prolactin ( PRL ) levels followed by a prolonged decrease in PRL secretion from the pituitary . While the inhibitory component of this biphasic response depends on the cannabinoid-induced activation of dopamine release from hypothalamic terminals located in the median eminence , the neurobiological mechanisms underlying the activation phase of PRL release remains to be explained . In the present study the possible direct effect of the cannabinoid receptor agonist DB00470 ( THC ) on prolactin secretion and DB02527 accumulation was examined in anterior pituitary cultures . THC ( 0.1 and 1 microM ) increased DB02527 levels , and induced PRL release ( 1 and 10 mu ) . THC did not affect vasoactive intestinal peptide ( P01282 , 0.5 microM ) induced DB02527 accumulation in pituitary cultures , showing additive effects at THC 1 microM concentration . However , THC did prevent P01282 -dependent increases in prolactin secretion . These results indicate that THC , through a direct pituitary action , activates both the synthesis of DB02527 and PRL release and interferes with intracellular mechanisms involved in PRL secretion by P01282 . These actions could be mediated through cannabinoid P21554 receptors which were found to be present in anterior pituitary cells , including lactotrophs , as revealed by immunocytochemistry with a specific polyclonal antibody raised against the P21554 receptor protein . Activation of large-conductance , Ca2+-activated K+ channels by cannabinoids . We have examined the effects of the cannabinoid anandamide ( AEA ) and its stable analog , methanandamide ( methAEA ) , on large-conductance , Ca2+-activated K+ ( BK ) channels using human embryonic kidney ( P29320 ) -293 cells , in which the alpha-subunit of the Q12791 ( BK-alpha ) , both alpha- and beta1-subunits ( BK-alphabeta1 ) , or both alpha- and beta4-subunits ( BK-alphabeta4 ) were heterologously expressed . In a whole cell voltage-clamp configuration , each cannabinoid activated BK-alphabeta1 within a similar concentration range . Because methAEA could potentiate BK-alpha , BK-alphabeta1 , and BK-alphabeta4 with similar efficacy , the beta-subunits may not be involved at the site of action for cannabinoids . Under cell-attached patch-clamp conditions , application of methAEA to the bathing solution increased Q12791 activity ; however , methAEA did not alter channel activity in the excised inside-out patch mode even when DB00171 was present on the cytoplasmic side of the membrane . Application of methAEA to P29320 -BK-alpha and P29320 -BK-alphabeta1 did not change intracellular Ca2+ concentration . Moreover , methAEA-induced potentiation of Q12791 currents was not affected by pretreatment with a P21554 antagonist ( AM251 ) , modulators of G proteins ( cholera and pertussis toxins ) or by application of a selective CB2 agonist ( JWH133 ) . Inhibitors of P62158 , PKG , and MAPKs ( W7 , KT5823 , and PD-98059 ) did not affect the potentiation . Application of methAEA to mouse aortic myocytes significantly increased Q12791 currents . This study provides the first direct evidence that unknown factors in the cytoplasm mediate the ability of endogenous cannabinoids to activate Q12791 currents . Cannabinoids may be hyperpolarizing factors in cells , such as arterial myocytes , in which BK channels are highly expressed . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Antiemetic and motor-depressive actions of CP55,940 : cannabinoid P21554 receptor characterization , distribution , and G-protein activation . Dibenzopyran ( Delta(9)-tetrahydrocannabinol ) and aminoalkylindole [ R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrolol[1,2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl) methanone mesylate ; ( WIN55,212-2 ) ] cannabinoids suppress vomiting produced by cisplatin via cannabinoid CB(1) receptors . This study investigates the antiemetic potential of the " nonclassical " cannabinoid CP55,940 [ 1alpha,2beta-(R)-5alpha ] -(-)-5-(1,1-dimethyl)-2- [ 5-hydroxy-2-(3-hydroxypropyl) cyclohexyl-phenol ] against cisplatin-induced vomiting and assesses the presence and functionality of cannabinoid CB(1) receptors in the least shrew ( Cryptotis parva ) brain . CP55,940 ( 0.025-0.3 mg/kg ) reduced both the frequency of cisplatin-induced emesis ( ID(50)=0.025 mg/kg ) and the percentage of shrews vomiting ( ID(50)=0.09 mg/kg ) . CP55,940 also suppressed shrew motor behaviors ( ID(50)=0.06- 0.21 mg/kg ) at such doses . The antiemetic and motor-suppressant actions of CP55,940 were countered by SR141716A [ N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)- DB01213 -3-carboxamide ] , indicating both effects are cannabinoid CB(1) receptor-mediated . Autoradiographic studies with [ 3H ] -SR141716A and [ 35S ] -GTPgammaS binding revealed that the distribution of the cannabinoid CB(1) receptor and its activation pattern are similar to rodent brain and significant levels are present in brain loci ( e.g. , nucleus tractus solitarius ( P30990 ) ) that control emesis . The affinity rank order of structurally diverse cannabinoid ligands for cannabinoid CB(1) receptor in shrew brain is similar to rodent brain : HU-210=CP55,940=SR141716A >/= WIN55,212-2 >/= DB00470 > methanandamide=HU-211=cannabidiol=2-arachidonoylglycerol . This affinity order is also similar and is highly correlated to the cannabinoid EC(50) potency rank order for GTPgammaS stimulation except WIN55,212-2 and DB00470 potency order were reversed . The affinity and the potency rank order of tested cannabinoids were significantly correlated with their antiemetic ID(50) potency order against cisplatin-induced vomiting ( CP55,940 > WIN55,212-2= DB00470 ) as well as emesis produced by 2-arachidonoylglycerol or SR141716A ( CP55,940 > WIN55,212-2 > DB00470 ) . Depolarization-induced suppression of excitation in murine autaptic hippocampal neurones . Depolarization-induced suppression of excitation and inhibition ( Q9UL01 and DSI ) appear to be important forms of short-term retrograde neuronal plasticity involving endocannabinoids ( eCB ) and the activation of presynaptic cannabinoid P21554 receptors . We report here that P21554 -dependent Q9UL01 can be elicited from autaptic cultures of excitatory mouse hippocampal neurones . Q9UL01 in autaptic cultures is both more robust and elicited with a more physiologically relevant stimulus than has been thus far reported for conventional hippocampal cultures . An additional requirement for autaptic Q9UL01 is filled internal calcium stores . Pharmacological experiments favour a role for 2-arachidonyl glycerol ( 2-AG ) rather than arachidonyl ethanolamide ( AEA ) or noladin ether as the relevant endocannabinoid to elicit Q9UL01 . In particular , the latter two compounds fail to reversibly inhibit EPSCs , a quality inconsistent with the role of bona fide eCB mediating Q9UL01 . Delta9- DB00470 ( delta9-THC ) fails to inhibit EPSCs , yet readily occludes both Q9UL01 and EPSC inhibition by a synthetic P21554 agonist , Q08050 55212-2 . With long-term exposure ( approximately 18 h ) , delta9-THC also desensitizes P21554 receptors . Lastly , a functional endocannabinoid transporter is necessary for the expression of Q9UL01 . CB(1) and CB(2) cannabinoid receptors mediate different aspects of DB00470 ( THC ) -induced T helper cell shift following immune activation by Legionella pneumophila infection . Legionella pneumophila infection of mice induces proinflammatory cytokines and Th1 immunity as well as rapid increases in serum levels of IL-12 and IFNgamma and splenic IL-12Rbeta2 expression . Delta-9-tetrahydrocannabinol ( THC ) treatment prior to infection causes a shift from Th1 to Th2 immunity and here we demonstrate that CB(1) and CB(2) cannabinoid receptors mediate different aspects of the shift . Using cannabinoid receptor antagonists and cannabinoid receptor gene deficient mice ( CB(1) ( -/- ) and CB(2) ( -/- ) ) , we showed that both CB(1) and CB(2) receptors were involved in the THC-induced attenuation of serum IL-12 and IFNgamma . IFNgamma production is dependent upon signaling through IL-12Rbeta2 ( beta2 ) and THC treatment suppressed splenic beta2 message ; moreover , this effect was CB(1) but not CB(2)-dependent from studies with receptor antagonists and P21554 (-/-) and CB2(-/-) mice . Furthermore , observed increases in P05112 induced by THC , were not involved in the drug effect on beta2 from studies with P05112 deficient mice . The GATA-3 transcription factor is necessary for P05112 production and is selectively expressed in Th2 cells . GATA-3 message levels were elevated in spleens of THC-treated and L. pneumophila-infected mice and the effect was shown to be CB(2) but not CB(1)-dependent . Furthermore , GATA-3 regulatory factors were modulated in that Notch ligand Q9NR61 mRNA was decreased and P78504 increased by THC also in a CB2-dependent manner and splenic NFkappaB p65 was increased . Together , these results indicate that CB(1) and CB(2) mediate the THC-induced shift in T helper activity in L. pneumophila-infected mice , with CB(1) involved in suppressing IL-12Rbeta2 and CB(2) involved in enhancing GATA-3 . The influence of costimulation and regulatory P01730 + T cells on intestinal IgA immune responses . It is thought that IgA B-cell differentiation is highly dependent on activated P01730 + T cells . In particular , cell-cell interactions in the Peyer 's patches involving P25942 and/or P33681 / P42081 have been implicated in germinal-center formation and IgA B-cell development . Also soluble factors , such as P05112 , P05113 , P05231 , and TGF beta may be critical for IgA B-cell differentiation in vivo . Here we report on some paradoxical findings with regard to IgA B-cell differentiation and specific mucosal immune responses that we have recently made using gene knockout mice . More specifically , we have investigated to what extent absence of P01730 + T cells , relevant cytokines , or T-cell-B-cell interactions would influence IgA B-cell differentiation in vivo . Using P01730 - or P05112 -gene knockout mice or mice made transgenic for DB01281 , we found that , although specific responses were impaired , total IgA production and IgA B-cell differentiation appeared to proceed normally . However , a poor correlation was found between , on the one hand , GC formation and IgA differentiation and , on the other hand , the ability to respond to T-cell-dependent soluble protein antigens in these mice . Thus , despite the various deficiencies in P01730 + T-cell functions seemingly intact IgA B-cell development was observed . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Role of endogenous cannabinoids in synaptic signaling . Research of cannabinoid actions was boosted in the 1990s by remarkable discoveries including identification of endogenous compounds with cannabimimetic activity ( endocannabinoids ) and the cloning of their molecular targets , the P21554 and CB2 receptors . Although the existence of an endogenous cannabinoid signaling system has been established for a decade , its physiological roles have just begun to unfold . In addition , the behavioral effects of exogenous cannabinoids such as DB00470 , the major active compound of hashish and marijuana , await explanation at the cellular and network levels . Recent physiological , pharmacological , and high-resolution anatomical studies provided evidence that the major physiological effect of cannabinoids is the regulation of neurotransmitter release via activation of presynaptic P21554 receptors located on distinct types of axon terminals throughout the brain . Subsequent discoveries shed light on the functional consequences of this localization by demonstrating the involvement of endocannabinoids in retrograde signaling at GABAergic and glutamatergic synapses . In this review , we aim to synthesize recent progress in our understanding of the physiological roles of endocannabinoids in the brain . First , the synthetic pathways of endocannabinoids are discussed , along with the putative mechanisms of their release , uptake , and degradation . The fine-grain anatomical distribution of the neuronal cannabinoid receptor P21554 is described in most brain areas , emphasizing its general presynaptic localization and role in controlling neurotransmitter release . Finally , the possible functions of endocannabinoids as retrograde synaptic signal molecules are discussed in relation to synaptic plasticity and network activity patterns . Opposing control of cannabinoid receptor stimulation on amyloid-beta-induced reactive gliosis : in vitro and in vivo evidence . Beside cytotoxic mechanisms impacting on neurons , amyloid beta ( A beta ) -induced astroglial activation is operative in Alzheimer 's disease brain , suggesting that persistent inflammatory response may have a role in the illness and that positive results may be achieved by curbing the astroglial reaction . Because the role of the endocannabinoid system could represent a promising field of research , the present study conducted in vitro and in vivo experiments to assess this system . P13671 rat astroglioma cells were challenged with 1 microg/ml A beta 1-42 in the presence or absence of selective agonists and antagonists of cannabinoid (CB)1 and CB2 receptors . Furthermore , rats were inoculated into the frontal cortex with 30 ng of A beta 1-42 and were i.p. administered with 5 mg/kg of the same substances . Immunohistochemical and biochemical findings revealed that selective agonism at P21554 and antagonism at CB2 receptors was able to blunt A beta-induced reactive astrogliosis with subsequent overexpression of glial fibrillary acidic protein and P04271 protein . Moreover , A beta provoked down-regulation of P21554 receptors together with a reduction of anandamide concentration , whereas CB2 receptors were up-regulated and 2-arachidonoyl glycerol concentration was increased . Finally , to our knowledge , the current study is the first showing that interactions at cannabinoid receptors result in a dual regulation of A beta-induced reactive astrogliosis . The data support the assumption that compounds able to selectively block CB2 receptors may have therapeutic potential in controlling A beta-related pathology , due to their beneficial effects devoid of psychotropic consequences . Construction of a steric map of the binding pocket for cannabinoids at the cannabinoid receptor . In order to gain information about the topology of the brain cannabinoid receptor ( P21554 ) , a Receptor Steric ( RS ) Map for cannabinoids at this receptor was calculated . The classical cannabinoids (-)-11-hydroxy- DB00470 ( P04264 = 210 +/- 56 nM ) , (-)-9-nor-9-beta-hydroxy-hexahydrocannabinol ( P04264 = 124 +/- 17 nM ) , nabilone ( P04264 = 120 +/- 13 nM ) , and the non-classical cannabinoid , CP-55,244 ( P04264 = 1.4 +/- .3 nM ) were used as template molecules . The RS map was obtained as the union of the van der Waals ' volumes of only those accessible conformers identified by P08253 calculations that were able to clear a region of steric interference at the P21554 receptor previously characterized by us [ Reggio , P.H. , Panu , A.M. and Miles , S. ( 1993 ) , J. Med. Chem. , 36 , 1761-1771 ] . The utility of the RS Map was explored by screening the accessible conformers of the classical cannabinoid , cannabinol ( CBN ) , ( P04264 = 3200 +/- 450 nM ) , for its ability to fit within the RS map . Only the global minimum energy conformer of CBN ( 53.2 % abundance at 298K ) was able to fit within the RS map . These results imply that one reason for the reduced affinity of CBN may be that only 53.2 % of CBN molecules are shaped properly to fit in the binding pocket for cannabinoids at the P21554 receptor . Cannabinoids against pain . Efficacy and strategies to reduce psychoactivity : a clinical perspective . The clinical use of cannabinoids is currently a topic of interest not exclusively , but most importantly , concerning different areas of pain therapy . One of the major obstacles in developing clinically acceptable compounds is the cannabimimetic side-effect profile of DB00470 ( THC ) and other cannabinoids . This article gives a brief overview of the endocannabinoid system , its components and functions and explains the current approaches to avoiding cannabimimetic side effects by separating them from the therapeutic effects . One of these approaches is the addition of cannabidiol ( DB09061 ) as well as the use of preparations suitable for oromucosal application . Also cannabinoids , which primarily stimulate peripheral cannabinoid-1 ( P21554 ) receptors or selectively cannabinoid-2 ( CB2 ) receptors , can further separate analgesic activity from cannabimimetic activity . Local or topical modes of application are another attempt aiming in the same direction . Modulating the endogenous cannabinoid tone ( via the inhibition of endocannabinoid-metabolising enzymes ) is another strategy . The combination of THC in low , non-psychoactive doses with opioids has a synergistic effect and reduces opioid tolerance effects . Available data from these approaches are summarised and their more and less promising aspects are discussed . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . Synthetic anti- Q96RJ3 antibodies that mimic Q9Y275 binding and target both human and murine B cells . Q96RJ3 , which is expressed on all mature B cells , is a specific receptor for the B-cell survival and maturation factor Q9Y275 ( Q9Y275 belonging to the tumor necrosis factor [ P01375 ] family ) . In order to investigate the consequences of targeting Q96RJ3 in murine models and to assess the potential of Q96RJ3 antibodies as human therapeutics , synthetic antibody phage libraries were employed to identify Q9Y275 -blocking antibodies cross-reactive to murine and human Q96RJ3 , which share 52 % identity in their extracellular domains . We found an antibody , P21554 , which exhibits muM affinity for murine Q96RJ3 and very weak affinity for the human receptor . CB3s , an affinity-matured variant of P21554 , has sub-nM affinity for Q96RJ3 from both species . DB00160 scanning and crystallographic structural analysis of the CB3s/ Q96RJ3 complex reveal that CB3s mimics Q9Y275 by interacting with a similar region of the Q96RJ3 surface . Despite this similarity in binding epitopes , P21554 variants antagonize Q9Y275 -dependent human B-cell proliferation in vitro and are effective at reducing murine B-cell populations in vivo , showing significant promise as therapeutics for human B-cell-mediated diseases . Sex-specific alterations in hippocampal cannabinoid 1 receptor expression following adolescent DB00470 treatment in the rat . Marijuana use by adolescents has been on the rise since the early 1990s . With recent legalization and decriminalization acts passed , cannabinoid exposure in adolescents will undoubtedly increase . Human studies are limited in their ability to examine underlying changes in brain biochemistry making rodent models valuable . Studies in adult and adolescent animals show region and sex specific downregulation of the cannabinoid 1 ( P21554 ) receptor following chronic cannabinoid treatment . However , although sex-dependent changes in behavior have been observed during the drug abstinence period following adolescent cannabinoid exposure , little is known about P21554 receptor expression during this critical time . In order to characterize P21554 receptor expression following chronic adolescent Δ-9-tetrahydrocannabinol ( THC ) exposure , we used [ (3)H ] CP55,940 binding to assess P21554 receptor expression in the dentate gyrus and areas P00915 , P00918 , and P07451 of the hippocampus in both male and female adolescent rats at both 24h and 2 weeks post chronic THC treatment . Consistent with other reported findings , we found downregulation of the P21554 receptor in the hippocampal formation at 24h post treatment . While this downregulation persisted in both sexes following two weeks of abstinence in the P00918 region , in females , this downregulation also persisted in areas P00915 and P07451 . Expression in the dentate gyrus returned to the normal range by two weeks . These data suggest that selective regions of the hippocampus show persistent reductions in P21554 receptor expression and that these reductions are more widespread in female compared to male adolescents . CB2 receptors in the brain : role in central immune function . Recently , it has been recognized that the cannabinoid receptor CB2 may play a functionally relevant role in the central nervous system ( CNS ) . This role is mediated primarily through microglia , a resident population of cells in the CNS that is morphologically , phenotypically , and functionally related to macrophages . These cells also express the cannabinoid receptor P21554 . The P21554 receptor ( CB1R ) is constitutively expressed at low levels while the CB2 receptor ( CB2R ) is expressed at higher levels and is modulated in relation to cell activation state . The relatively high levels of the CB2R correspond with microglia being in ' responsive ' and ' primed ' states , suggesting the existence of a ' window ' of functional relevance during which activation of the CB2R modulates microglial activities . Signature activities of ' responsive ' and ' primed ' microglia are chemotaxis and antigen processing , respectively . The endocannabinoid 2-arachidonylglycerol has been reported to stimulate a chemotactic response from these cells through the CB2R . In contrast , we have shown in vivo and in vitro that the exogenous cannabinoids DB00470 and CP55940 inhibit the chemotactic response of microglia to Acanthamoeba culbertsoni , an opportunistic pathogen that is the causative agent of Granulomatous Amoebic Encephalitis , through activation of the CB2R . It is postulated that these exogenous cannabinoids superimpose an inhibitory effect on pro-chemotactic endocannabinoids that are elicited in response to Acanthamoeba . Furthermore , the collective results suggest that the CB2R plays a critical immune functional role in the CNS . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Targeting CB2 cannabinoid receptors as a novel therapy to treat malignant lymphoblastic disease . In the current study , we examined whether ligation of CB2 receptors would lead to induction of apoptosis in tumors of immune origin and whether CB2 agonist could be used to treat such cancers . Exposure of murine tumors EL-4 , LSA , and P815 to DB00470 ( THC ) in vitro led to a significant reduction in cell viability and an increase in apoptosis . Exposure of EL-4 tumor cells to the synthetic cannabinoid HU-210 and the endogenous cannabinoid anandamide led to significant induction of apoptosis , whereas exposure to WIN55212 was not effective . Treatment of EL-4 tumor-bearing mice with THC in vivo led to a significant reduction in tumor load , increase in tumor-cell apoptosis , and increase in survival of tumor-bearing mice . Examination of a number of human leukemia and lymphoma cell lines , including Jurkat , Molt-4 , and Sup-T1 , revealed that they expressed CB2 receptors but not P21554 . These human tumor cells were also susceptible to apoptosis induced by THC , HU-210 , anandamide , and the CB2-selective agonist JWH-015 . This effect was mediated at least in part through the CB2 receptors because pretreatment with the CB2 antagonist SR144528 partially reversed the THC-induced apoptosis . Culture of primary acute lymphoblastic leukemia cells with THC in vitro reduced cell viability and induced apoptosis . Together , the current data demonstrate that CB2 cannabinoid receptors expressed on malignancies of the immune system may serve as potential targets for the induction of apoptosis . Also , because CB2 agonists lack psychotropic effects , they may serve as novel anticancer agents to selectively target and kill tumors of immune origin . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . ∆(9)- DB00470 decreases O60936 receptor density and mRNA levels in human SH-SY5Y cells . Several studies demonstrated a cross-talk between the opioid and cannabinoid system . The O60936 receptor and its endogenous ligand nociceptin/orphanin FQ represent an opioid-related functional entity that mediates some non-classical opioid effects . The relationship between cannabinoid and nociceptin/ O60936 system is yet poorly explored . In this study , we used the neuroblastoma SH-SY5Y cell line to investigate the effect of DB00470 ( ∆(9)-THC ) on nociceptin/ O60936 system . Results revealed that the exposure to ∆(9)-THC ( 100 , 150 , and 200 nM ) for 24 h produces a dose-dependent O60936 receptor B ( max ) down-regulation . Moreover , ∆(9)-THC caused a dose-dependent decrease in O60936 mRNA levels . The selective cannabinoid receptor P21554 antagonist AM251 ( 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1-piperidinyl-1H-pyrazole-3-carboxamide ) reduces both effects , suggesting that ∆(9)-THC activation of P21554 receptor is involved in the observed effects . These data show evidence of a cross-talk between O60936 and P21554 receptors , thus suggesting a possible interplay between cannabinoid and nociceptin/ O60936 system . Delta(9)- DB00470 enhances MCF-7 cell proliferation via cannabinoid receptor-independent signaling . We recently reported that Delta(9)-tetrahydrocannabinol ( Delta(9)-THC ) has the ability to stimulate the proliferation of human breast carcinoma MCF-7 cells . However , the mechanism of action remains to be clarified . The present study focused on the relationship between receptor expression and the effects of Delta(9)-THC on cell proliferation . RT-PCR analysis demonstrated that there was no detectable expression of CB receptors in MCF-7 cells . In accordance with this , no effects of cannabinoid 1/2 ( P21554 /2 ) receptor antagonists and pertussis toxin on cell proliferation were observed . Although MCF-7 cell proliferation is suggested to be suppressed by Delta(9)-THC in the presence of CB receptors , it was revealed that Delta(9)-THC could exert upregulation of living cells in the absence of the receptors . Interestingly , Delta(9)-THC upregulated human epithelial growth factor receptor type 2 ( P04626 ) expression , which is known to be a predictive factor of human breast cancer and is able to stimulate cancer cells as well as MCF-7 cells . DB00970 -treatment interfered with the upregulation of P04626 and cell proliferation by cannabinoid . Taken together , these studies suggest that , in the absence of CB receptors , Delta(9)-THC can stimulate the proliferation of MCF-7 cells by modulating , at least in part , P04626 transcription . Cerebrospinal anandamide levels are elevated in acute schizophrenia and are inversely correlated with psychotic symptoms . The endocannabinoids are a family of bioactive lipids that activate P21554 cannabinoid receptors in the brain and exert intense emotional and cognitive effects . Here , we have examined the role of endocannabinoid signaling in psychotic states by measuring levels of the endocannabinoid anandamide in cerebrospinal fluid ( P04141 ) of acute paranoid-type schizophrenic patients . We found that P04141 anandamide levels are eight-fold higher in antipsychotic-naive first-episode paranoid schizophrenics ( n = 47 ) than healthy controls ( n = 84 ) , dementia patients ( n = 13 ) or affective disorder patients ( n = 22 ) . Such an alteration is absent in schizophrenics treated with ' typical ' antipsychotics ( n = 37 ) , which antagonize dopamine D2-like receptors , but not in those treated with ' atypical ' antipsychotics ( n = 34 ) , which preferentially antagonize 5HT(2A) receptors . Furthermore , we found that , in nonmedicated acute schizophrenics , P04141 anandamide is negatively correlated with psychotic symptoms ( rS = -0.452 , P = 0.001 ) . The results suggest that anandamide elevation in acute paranoid schizophrenia may reflect a compensatory adaptation to the disease state . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Effects of DB00470 and ( R ) -methanandamide on open-field behavior in rats . This study compared the effects of ( R ) -methanandamide , an analog of the mammalian brain constituent anandamide , and DB00470 on the open-field behavior of male Sprague-Dawley rats . Rats were individually housed with free access to food and water . Animals were treated with 0 , 1 , 3 , and 5.6 mg/kg DB00470 given i.p . 30 min pre-session ; and 0 , 3 , 10 , and 18 mg/kg ( R ) -methanandamide , 15 min pre-session . The behavioral categories recorded were ambulation ( the number of squares crossed ) , rearing ( the number of times the rat stood erect on its hind-legs ) , latency ( the time in seconds to leave the starting area , the circle in the center of the field ) , circling ( the number of times the animal turned around its vertical axis , 0.5 point given for each 180 degrees turn ) , grooming ( the number of cleaning bouts ) , urination and defecation ( the number of urination spots and fecal boli deposited during the 5 min observation period ) . Delta-9-tetrahydrocannabinol was more potent than ( R ) -methanandamide , but otherwise the effects of DB00470 and ( R ) -methanandamide were similar , with one exception ; whereas DB00470 produced dose-related increases in circling , ( R ) -methanandamide did not increase circling over the doses examined . The DB00470 -induced increase in circling was blocked by the central cannabinoid receptor P21554 antagonist SR 141716 . The differential effects with regard to circling may indicate that there are qualitative behavioral differences in the effects of DB00470 and ( R ) -methanandamide . The revised human liver cytochrome P450 " Pie " : absolute protein quantification of CYP4F and CYP3A enzymes using targeted quantitative proteomics . The CYP4F subfamily of enzymes has been identified recently to be involved in the metabolism of endogenous compounds ( arachidonic acid and leukotriene B4 ) , nutrients ( vitamins P04264 and E ) , and xenobiotics ( pafuramidine and fingolimod ) . P78329 and CYP4F3B are reported to be expressed in the human liver . However , absolute concentrations of these enzymes in human liver microsomes ( HLMs ) and their interindividual variability have yet to be determined because of the lack of specific antibodies . Here , an liquid chromatography with tandem mass spectrometry ( LC-MS/MS ) -based targeted quantitative proteomic approach was employed to determine the absolute protein concentrations of P78329 and CYP4F3B compared with CYP3A in two panels of HLMs ( n = 31 ) . As a result , the human hepatic cytochrome P450 ( P450 ) " pie " has been revised to include the contribution of CYP4F enzymes , which amounts to 15 % of the total hepatic cytochrome P450 enzymes . CYP4F3B displayed low interindividual variability ( 3.3-fold ) in the HLM panels whereas P78329 displayed large variability ( 21-fold ) . However , P78329 variability decreased to 3.4-fold if the two donors with the lowest expression were excluded . In contrast , CYP3A exhibited 29-fold interindividual variability in the same HLM panels . The proposed marker reaction for CYP4F enzymes pafuramidine/DB289 M1 formation did not correlate with CYP4F protein content , suggesting alternate metabolic pathways for DB289 M1 formation in HLMs . In conclusion , CYP4F enzymes are highly expressed in the human liver and their physiologic and pharmacologic roles warrant further investigation . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . The influence of mast cell mediators on migration of SW756 cervical carcinoma cells . The role of mast cell mediators on cervical cancer cell migration was assessed using an in vitro assay of scratch wound healing onto monolayers of HPV18-positive cervical carcinoma cells ( SW756 ) . Migration of SW756 cells was accelerated by co-culture with the mast cell line LAD2 . This effect was inhibited by the P35367 antagonist DB06691 and the cannabinoid agonists 2-arachidonylglycerol ( 2AG ) and Win 55,212-2 . Therefore , the specific effects of histamine and cannabinoids on SW756 migration and LAD2 activation were analyzed . DB11320 added to the in vitro assay of scratch wound healing either increased or inhibited SW756 migration rate by acting either on P35367 or Q9H3N8 , respectively . Cannabinoids acted on P21554 receptors to inhibit SW756 migration . Supernatants from SW756 cells stimulated LAD2 cell degranulation , which in turn was inhibited by cannabinoids acting via CB2 receptors . RT-PCR showed that SW756 expressed mRNA for P21554 , CB2 , P35367 , P25021 , and Q9H3N8 . On the other hand , LAD2 expressed mRNA for all four HRs and CB2 . The results suggest that mast cells could be contributing to cervical cancer cell invasion and spreading by the release of histamine and cannabinoids . Therefore , therapeutic modulation of specific mast cell mediators may be beneficial for cervical cancer treatment . Additive antiemetic efficacy of low-doses of the cannabinoid CB(1/2) receptor agonist Δ(9)-THC with ultralow-doses of the vanilloid Q8NER1 receptor agonist resiniferatoxin in the least shrew ( Cryptotis parva ) . Previous studies have shown that cannabinoid P21554 /2 and vanilloid Q8NER1 agonists ( DB00470 ( Δ(9)-THC ) and resiniferatoxin ( RTX ) , respectively ) can attenuate the emetic effects of chemotherapeutic agents such as cisplatin . In this study we used the least shrew to demonstrate whether combinations of varying doses of Δ(9)-THC with resiniferatoxin can produce additive antiemetic efficacy against cisplatin-induced vomiting . RTX by itself caused vomiting in a bell-shaped dose-dependent manner with maximal vomiting at 18 μg/kg when administered subcutaneously ( s.c. ) but not intraperitoneally ( i.p. ) . Δ(9)-THC up to 10 mg/kg provides only 80 % protection of least shrews from cisplatin-induced emesis with an ID50 of 0.3-1.8 mg/kg . Combinations of 1 or 5 μg/kg RTX with varying doses of Δ(9)-THC completely suppressed both the frequency and the percentage of shrews vomiting with ID50 dose values 5-50 times lower than Δ(9)-THC doses tested alone against cisplatin . A less potent Q8NER1 agonist , capsaicin , by itself did not cause emesis ( i.p. or s.c. ) , but it did significantly reduce vomiting induced by cisplatin given after 30 min but not at 2 h . The Q8NER1 -receptor antagonist , ruthenium red , attenuated cisplatin-induced emesis at 5mg/kg ; however , another Q8NER1 -receptor antagonist , capsazepine , did not . In summary , we present evidence that combination of P21554 /2 and Q8NER1 agonists have the capacity to completely abolish cisplatin-induced emesis at doses that are ineffective when used individually . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Creating a genotype-based dosing algorithm for acenocoumarol steady dose . DB01418 is a commonly prescribed anticoagulant drug for the prophylaxis and treatment of venous and arterial thromboembolic disorders in several countries . In counterpart of warfarin , there is scarce information about pharmacogenetic algorithms for steady acenocoumarol dose estimation . The aim of this study was to develop an algorithm of prediction for acenocoumarol.The algorithm was created using the data from 973 retrospectively selected anticoagulated patients and was validated in a second independent cohort adding up to 2,683 patients . The best regression model to predict stable dosage in the Primary Cohort included clinical factors ( age and body mass index , BSA ) and genetic variants ( Q9BQB6 , P11712 * and P78329 polymorphisms ) and explained up to 50 % of stable dose . In the validation study the clinical algorithm yielded an adjusted R²=0.15 ( estimation´s standard error=4.5 ) and the genetic approach improved the dose forecast up to 30 % ( estimation´s standard error=4.6 ) . Again , the best model combined clinical and genetic factors ( R² = 0.48 ; estimation´s standard error=4 ) which provided the best results of doses estimates within 20 % of the real dose in patients taking lower ( ≤ 7 mg/week ) or higher ( ≥ 25 mg/week ) acenocoumarol doses . In conclusion , we developed a prediction algorithm using clinical data and three polymorphisms in Q9BQB6 , P11712 * and P78329 genes that provided a steady acenocoumarol dose for about 50 % of patients in the Validation Cohort . Such algorithm was especially useful to patients who need higher or lower acenocoumarol doses , those patients with higher time required until their stabilisation and are more prone to suffer a treatment derived complication . Cannabinoid physiology and pharmacology : 30 years of progress . Delta9- DB00470 from Cannabis sativa is mimicked by cannabimimetic analogs such as CP55940 and WIN55212-2 , and antagonized by rimonabant and SR144528 , through G-protein-coupled receptors , P21554 in the brain , and CB2 in the immune system . Eicosanoids anandamide and 2-arachidonoylglycerol are the " endocannabinoid " agonists for these receptors . P21554 receptors are abundant in basal ganglia , hippocampus and cerebellum , and their functional activity can be mapped during behaviors using cerebral metabolism as the neuroimaging tool . P21554 receptors couple to G(i/o) to inhibit DB02527 production , decrease Ca2+ conductance , increase K+ conductance , and increase mitogen-activated protein kinase activity . Functional activation of G-proteins can be imaged by [35S]GTPgammaS autoradiography . Post-synaptically generated endocannabinoids form the basis of a retrograde signaling mechanism referred to as depolarization-induced suppression of inhibition ( DSI ) or excitation ( Q9UL01 ) . Under circumstances of sufficient intracellular Ca2+ ( e.g. , burst activity in seizures ) , synthesis of endocannabinoids releases a diffusible retrograde messenger to stimulate presynaptic P21554 receptors . This results in suppression of gamma-aminobutyric acid ( GABA ) release , thereby relieving the post-synaptic inhibition . Tolerance develops as neurons adjust both receptor number and cellular signal transduction to the chronic administration of cannabinoid drugs . Future therapeutic drug design can progress based upon our current understanding of the physiology and pharmacology of P21554 , CB2 and related receptors . One very important role for P21554 antagonists will be in the treatment of craving in the disease of substance abuse . delta 9- DB00470 increases activity of tyrosine hydroxylase in cultured fetal mesencephalic neurons . The exposure of pregnant rats to delta 9-tetrahydrocannabinol ( delta 9-THC ) , the main psychoactive constituent of Cannabis sativa , during gestation and lactation , affects the gene expression and the activity of tyrosine hydroxylase ( TH ) in the brain of their offspring , measured at fetal and early postnatal ages , when the expression of this enzyme plays an important role in neural development . In the present article , we have examined whether delta 9-THC is able to affect TH activity in cultured mesencephalic neurons obtained from fetuses at gestational d 14 . Thus , TH activity increased approximately twofold in cells obtained from naive fetuses when exposed for 24 h to medium containing delta 9-THC . In addition , TH activity was also approx twofold higher in cells obtained from fetuses exposed daily to delta 9-THC from d 5 of gestation than in cells obtained from control fetuses , when both were exposed to basal media . This effect of delta 9-THC on TH activity seems to be produced via the activation to cannabinoid receptors , in particular the P21554 subtype , which would presumably be located in these cells . This is because the exposure to medium containing both delta 9-THC and SR141716A , a specific antagonist for P21554 receptors , abolished the effect observed with delta 9-THC alone . SR141716A alone was without effect on TH activity . Collectively , our results support the notion that delta 9-THC increased TH activity in cultured mesencephalic neurons , as previously observed in vivo , and that this effect was produced by activation of P21554 receptors , which seem to be operative at these early ages . All this points to a role for the endogenous cannabimimetic system in brain development . Identification of a high-affinity binding site involved in the transport of endocannabinoids . Phytocannabinoids , such as the principal bioactive component of marijuana , delta9-tetrahydrocannabinol , have been used for thousands of years for medical and recreational purposes . DB00470 and endogenous cannabinoids ( e.g. , anandamide ) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors ( P21554 and CB2 ) . The biosynthesis and physiology of anandamide is well understood , but its mechanism of uptake ( resulting in signal termination by fatty acid amide hydrolase ) has been elusive . Mounting evidence points to the existence of a specific anandamide transport protein ; however , no direct evidence for this protein has been provided . Here , we use a potent , competitive small molecule inhibitor of anandamide uptake ( LY2318912 , IC50 7.27 +/- 0.510 nM ) to identify a high-affinity , saturable anandamide transporter binding site ( LY2318912 ; K(d) = 7.62 +/- 1.18 nM , B(max) = 31.6 +/- 1.80 fmol/mg protein ) that is distinct from fatty acid amide hydrolase . Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function . Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling , and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors . The role of the P21554 receptor in the regulation of sleep . During the 1990s , transmembranal proteins in the central nervous system ( CNS ) that recognize the principal compound of marijuana , the DB00470 ( Delta9-THC ) were described . The receptors were classified as central or peripheral , P21554 and CB2 , respectively . To this date , it has been documented the presence in the CNS of specific lipids that bind naturally to the P21554 /CB2 receptors . The family of endogenous cannabinoids or endocannabinoids comprises oleamide , arachidonoylethanolamine , 2-arachidonylglycerol , virodhamine , noladin ether and N-arachidonyldopamine . Pharmacological experiments have shown that those compounds induce cannabimimetic effects . Endocannabinoids are fatty acid derivates that have a variety of biological actions , most notably via activation of the cannabinoid receptors . The endocannabinoids have an active role modulating diverse neurobiological functions , such as learning and memory , feeding , pain perception and sleep generation . Experimental evidence shows that the administration of Delta9-THC promotes sleep . The activation of the P21554 receptor leads to an induction of sleep , this effect is blocked via the selective antagonist . Since the system of the endogenous cannabinoids is present in several species , including humans , this leads to the speculation of the neurobiological role of the endocannabinoid system on diverse functions such as sleep modulation . This review discusses the evidence of the system of the endocannabinoids as well as their physiological role in diverse behaviours , including the modulation of sleep . Cannabinoid receptors in acute and chronic complications of atherosclerosis . Atherosclerosis is a chronic inflammatory disease that is the primary cause of myocardial infarction and stroke , which occur after sudden thrombotic occlusion of an artery . A growing body of evidence suggests that cannabinoid signalling plays a fundamental role in atherosclerosis development and its clinical manifestations . Thus , CB2 receptors are protective in myocardial ischaemia/reperfusion and implicated in the modulation of chemotaxis , which is crucial for the recruitment of leukocytes during inflammation . Delta-9- DB00470 ( THC ) -mediated activation has been shown to inhibit atherosclerotic plaque progression in a CB2 dependent manner . Although P21554 and CB2 expression has been reported on platelets , their involvement in thrombus formation is still controversial . While several reports suggest that P21554 receptors may have a relevant role in neuroprotection after ischaemic stroke , recent studies show the protective effects in various forms of neuroprotection are not related to P21554 stimulation , and a protective role of P21554 blockade has also been reported . In addition , vascular and myocardial P21554 receptors contribute to the modulation of blood pressure and heart rate . It is tempting to suggest that pharmacological modulation of the endocannabinoid system is a potential novel therapeutic strategy in the treatment of atherosclerosis . For these purposes , it is important to better understand the complex mechanisms of endocannabinoid signalling and potential consequences of its pharmacological modulation , as it may have both pro- and anti-atherosclerotic effects . Actions of DB00470 in cannabis : relation to use , abuse , dependence . Cannabis use disorders have been recently identified as a relevant clinical issue : a subset of cannabis smokers seeks treatment for their cannabis use , yet few succeed in maintaining long-term abstinence . The rewarding and positive reinforcing effects of the primary psychoactive component of smoked cannabis , DB00470 ( THC ) are mediated by the cannabinoid P21554 receptor . The P21554 receptor has also been shown to mediate cannabinoid dependence and expression of withdrawal upon cessation of drug administration , a phenomenon verified across species . This paper will review findings implicating the P21554 receptor in the behavioural effects of exogenous cannabinoids with a focus on cannabinoid dependence and reinforcement , factors that contribute to the maintenance of chronic cannabis smoking despite negative consequences . Opioidergic modulation of these effects is also discussed . Inhibition of recombinant human T-type calcium channels by Delta9-tetrahydrocannabinol and cannabidiol . Delta(9)- DB00470 ( THC ) and cannabidiol ( DB09061 ) are the most prevalent biologically active constituents of Cannabis sativa . THC is the prototypic cannabinoid P21554 receptor agonist and is psychoactive and analgesic . DB09061 is also analgesic , but it is not a P21554 receptor agonist . Low voltage-activated T-type calcium channels , encoded by the Ca(V)3 gene family , regulate the excitability of many cells , including neurons involved in nociceptive processing . We examined the effects of THC and DB09061 on human Ca(V)3 channels stably expressed in human embryonic kidney 293 cells and T-type channels in mouse sensory neurons using whole-cell , patch clamp recordings . At moderately hyperpolarized potentials , THC and DB09061 inhibited peak Ca(V)3.1 and Ca(V)3.2 currents with IC(50) values of approximately 1 mum but were less potent on Ca(V)3.3 channels . THC and DB09061 inhibited sensory neuron T-type channels by about 45 % at 1 mum . However , in recordings made from a holding potential of -70 mV , 100 nm THC or DB09061 inhibited more than 50 % of the peak Ca(V)3.1 current . THC and DB09061 produced a significant hyperpolarizing shift in the steady state inactivation potentials for each of the Ca(V)3 channels , which accounts for inhibition of channel currents . Additionally , THC caused a modest hyperpolarizing shift in the activation of Ca(V)3.1 and Ca(V)3.2 . THC but not DB09061 slowed Ca(V)3.1 and Ca(V)3.2 deactivation and inactivation kinetics . Thus , THC and DB09061 inhibit Ca(V)3 channels at pharmacologically relevant concentrations . However , THC , but not DB09061 , may also increase the amount of calcium entry following T-type channel activation by stabilizing open states of the channel . Effect of (-)-Delta(9)-tetrahydrocannabinoid on the hepatic redox state of mice . (-)-Delta(9)- DB00470 ( Delta(9)-THC ) , a psychoactive component of marijuana , has been reported to induce oxidative damage in vivo and in vitro . In this study , we administered Delta(9)-THC to healthy C57BL/6J mice aged 15 weeks in order to determine its effect on hepatic redox state . Mice were divided into 3 groups : Delta(9)-THC ( N = 10 ) , treated with 10 mg/kg body weight Delta(9)-THC daily ; VCtrl ( N = 10 ) , treated with vehicle [ 1:1:18 , cremophor EL ( polyoxyl 35 castor oil ) /ethanol/saline ] ; Ctrl ( N = 10 ) , treated with saline . Animals were injected ip twice a day with 5 mg/kg body weight for 10 days . Lipid peroxidation , protein carbonylation and DNA oxidation were used as biomarkers of oxidative stress . The endogenous antioxidant defenses analyzed were glutathione ( DB00143 ) levels as well as enzyme activities of superoxide dismutase , catalase , glutathione S-transferase , glutathione reductase , and glutathione peroxidase ( GPx ) in liver homogenates . The levels of mRNA of the cannabinoid receptors P21554 and CB2 were also monitored . Treatment with Delta(9)-THC did not produce significant changes in oxidative stress markers or in mRNA levels of P21554 and CB2 receptors in the liver of mice , but attenuated the increase in the selenium-dependent GPx activity ( Delta(9)-THC : 8 % ; VCtrl : 23 % increase ) and the DB00143 /oxidized DB00143 ratio ( Delta(9)-THC : 61 % ; VCtrl : 96 % increase ) , caused by treatment with the vehicle . Delta(9)-THC administration did not show any harmful effects on lipid peroxidation , protein carboxylation or DNA oxidation in the healthy liver of mice but attenuated unexpected effects produced by the vehicle containing ethanol/cremophor EL . (-)-Delta9-tetrahydrocannabinol antagonizes the peripheral cannabinoid receptor-mediated inhibition of adenylyl cyclase . (-)-Delta9- DB00470 ( (-)-Delta9-THC ) is the major active psychotropic component of the marijuana plant , Cannabis sativa . The membrane proteins that have been found to bind this material or its derivatives have been called the cannabinoid receptors . Two GTP-binding protein-coupled cannabinoid receptors have been cloned . P21554 or the neuronal cannabinoid receptor is found mostly in neuronal cells and tissues while CB2 or the peripheral cannabinoid receptor has been detected in spleen and in several cells of the immune system . It has previously been shown that activation of P21554 or CB2 receptors by cannabinoid agonists inhibits adenylyl cyclase activity . Utilizing Chinese hamster ovary cells and COS cells transfected with the cannabinoid receptors we report that (-)-Delta9-THC binds to both receptors with similar affinity . However , in contrast to its capacity to serve as an agonist for the P21554 receptor , (-)-Delta9-THC was only able to induce a very slight inhibition of adenylyl cyclase at the CB2 receptor . Morever , (-)-Delta9-THC antagonizes the agonist-induced inhibition of adenylyl cyclase mediated by CB2 . Therefore , we conclude that (-)-Delta9-THC constitutes a weak antagonist for the CB2 receptor . The beta2-adrenergic receptor specifically sequesters Gs but signals through both Gs and Gi/o in rat sympathetic neurons . Beta(2)-adrenergic receptors ( beta(2)-AR ) and P21554 cannabinoid receptors share the property of being constitutively active . The P21554 cannabinoid receptor can also sequester G(i/o) proteins ; however , it is not known whether the beta(2)-AR can also sequester G proteins . Beta(2)-ARs were heterologously expressed in rat superior cervical ganglion neurons by microinjection of cDNA and studied using the patch-clamp technique . The beta-AR agonist isoproterenol increased the Ca(2+) current 25.9+/-1.6 % in neurons microinjected with 100 ng/microl beta(2)-AR cDNA but was without effect on control neurons . Pretreatment with cholera toxin ( CTX ) abolished the effect of isoproterenol , indicating coupling via G(s) proteins . In neurons microinjected with 200 ng/microl beta(2)-AR cDNA , isoproterenol had the opposite effect of inhibiting the Ca(2+) current 36.5+/-2.0 % . Inhibition of the Ca(2+) current was sensitive to pertussis toxin , indicating beta(2)-AR coupling to G(i/o) proteins . Pretreatment with CTX resulted in a greater 54+/-3.8 % inhibition of the Ca(2+) current , indicating that G(s) coupling masks the full effect of G(i/o) coupling . Expression of beta(2)-ARs abolished signaling by G(s)-coupled receptors for vasoactive intestinal polypeptide ( P01282 ) . P01282 inhibited the Ca(2+) current 49.5+/-0.5 % in control neurons but had no effect in neurons expressing beta(2)-ARs . In contrast , expression of beta(2)-ARs had no effect on signaling by the G(i/o)-coupled alpha(2)-adrenergic receptor . This study demonstrates that the beta(2)-AR couples to both G(s) and G(i/o) proteins but specifically sequesters G(s) proteins , preventing their interaction with another G(s)-coupled receptor . beta(2)-adrenergic receptors thus have the potential to prevent other G(s)-coupled receptors from transducing their biological signals . Identification and characterization of a general nuclear translocation signal in signaling proteins . Upon stimulation , many proteins translocate into the nucleus in order to regulate a variety of cellular processes . The mechanism underlying the translocation is not clear since many of these proteins lack a canonical nuclear localization signal ( NLS ) . We searched for an alternative mechanism in extracellular signal-regulated kinase ( P29323 ) -2 and identified a 3 amino acid domain ( P49903 ) that is phosphorylated upon stimulation to induce nuclear translocation of P28482 . A 19 amino acid stretch containing this phosphorylated domain inserts nondiffusible proteins to the nucleus autonomously . The phosphorylated P49903 acts by binding to importin7 and the release from nuclear pore proteins . This allows its functioning both in passive and active P29323 transports . A similar domain appears in many cytonuclear shuttling proteins , and we found that phosphorylation of similar sequences in P84022 or Q02750 also induces their nuclear accumulation . Therefore , our findings show that this phosphorylated domain acts as a general nuclear translocation signal ( P30990 ) . P21554 receptor knockout mice show similar behavioral modifications to wild-type mice when enkephalin catabolism is inhibited . Behavioral and biochemical studies have suggested a functional link between the endogenous cannabinoid and opioid systems . Different hypotheses have been proposed to explain the interactions between opioid and cannabinoid systems such as a common pathway stimulating the dopaminergic system , a facilitation of signal-transduction- and/or a cannabinoid-induced enhancement of opioid peptide release . However , at this time , all the studies have been performed with exogenous agonists ( DB00470 or morphine ) , leading to a generally excessive stimulation of receptors normally stimulated by endogenous effectors ( anandamide or opioid peptides ) in various brain structures . To overcome this problem , we have measured various behavioral responses induced by the stimulation of the endogenous opioid system using the dual inhibitor of enkephalin-degrading enzymes , RB101 , in P21554 receptor knockout mice . Thus , analgesia , locomotor activity , anxiety and antidepressant-like effects were measured after RB101 administration ( 80 and 120 mg/kg i.p. or 10 mg/kg , i.v. ) in P21554 receptor knockout mice and their wild-type littermates . In all the experiments , inhibition of enkephalin catabolism produced similar modifications in behavior observed in P21554 knockout and wild-type mice . These results suggest limited physiological interaction between cannabinoid and opioid systems . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Inhibition of endocannabinoid neuronal uptake and hydrolysis as strategies for developing anxiolytic drugs . The endocannabinoid system comprises the P21554 and CB2 receptors ( the targets of the Cannabis sativa compound DB00470 ) , the endogenous ligands ( endocannabinoids ) arachidonoyl ethanolamide ( anandamide ) and 2-arachidonoyl glycerol , their synthesizing machinery and membrane transport system , and the hydrolyzing enzymes fatty acid amide hydrolase ( FAAH ) and monoacylglycerol lipase ( Q99685 ) , respectively . The endocannabinoids may act on demand to confer protection against aversive stimuli , which suggests that increasing their brain levels may represent an approach for treatment of anxiety-related disorders . Thus , this article reviews the profile of endocannabinoid reuptake and hydrolysis inhibitors in experimental tests predictive of anxiolytic activity . The FAAH inhibitors and the blockers of anandamide transport , in contrast to direct P21554 receptor agonists , induce anxiolytic effects at doses that do not interfere with motor activity . Q99685 inhibitors also reduce anxiety-like behavior , although they are more likely to impair motor activity . Regarding their mechanisms , increasing anandamide levels induce responses mediated by the P21554 receptor and occluded by the transient receptor potential vanilloid type-1 channels , whereas the effects of increasing 2-arachidonoyl glycerol depend on both P21554 and CB2 receptors . Their neuroanatomical targets include various structures related to anxiety and fear responses . Understanding the pharmacological properties of FAAH and Q99685 inhibitors may contribute toward the development of new anxiolytic interventions based on the endocannabinoid system . Possible involvement of endocannabinoids in the increase of morphine consumption in maternally deprived rat . Whether adolescent exposure to chronic DB00470 ( THC ) facilitates progression to opioid consumption is still controversial . In a maternal deprivation model ( 3 h daily from postnatal day 1-14 ) , we previously reported that adolescent exposure to chronic THC blocks morphine dependence in maternally deprived ( D ) rats . Owing to the existence of a functional cross-interaction between the opioid and cannabinoid systems in reward , we evaluated if the vulnerability to opiate reward in D rats , may involve an alteration of the endocannabinoid system . Anandamide and 2-arachidonoylglycerol ( 2-AG ) , were quantified in the striatum and mesencephalon of adolescent and adult D and non-deprived ( animal facility rearing , AFR ) rats by isotope dilution liquid chromatography-mass spectrometry . Oral morphine self-administration behavior was analyzed for 14 weeks , 24 days after chronic injection of the cannabinoid P21554 receptor antagonist/inverse agonist , SR141716A ( 3 mg/kg ) for 2 weeks during adolescence ( P01160 35-48 ) . Adolescent D rats exhibited higher basal levels of anandamide than adolescent AFR rats in the nucleus accumbens ( 38 % ) , the caudate-putamen nucleus ( 62 % ) and the mesencephalon ( 320 % ) , whereas adult D rats showed an increase of anandamide and 2-AG levels in the nucleus accumbens ( 50 % and 24 % , respectively ) and of 2-AG in the caudate-putamen nucleus ( 48 % ) , compared to adult AFR rats . Chronic administration of SR141716A to adolescent D rats blocked the escalation behavior in the morphine consumption test . Our data suggest that altered brain endocannabinoid levels may contribute to the escalation behavior in the morphine consumption test in a maternal deprivation model . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Transforming growth factor-beta and ciliary neurotrophic factor synergistically induce vasoactive intestinal peptide gene expression through the cooperation of Smad , P35610 , and AP-1 sites . The cytokine ciliary neurotrophic factor ( P26441 ) and transforming growth factor-beta ( TGF-beta ) both induce transcription of the vasoactive intestinal peptide ( P01282 ) gene through a 180-base pair cytokine response element ( CyRE ) in the P01282 promoter . While P26441 induces P35610 and AP-1 proteins to bind to cognate sites in the P01282 CyRE , the mechanism through which TGF-beta acts to induce P01282 gene transcription is not known . Here we show that P84022 and Q13485 proteins can bind to two distinct sites within the P01282 CyRE . These sites are absolutely required for the induction of P01282 CyRE transcription by TGF-beta . TGF-beta induces endogenous Smad-containing complexes to bind to these sites in human neuroblastoma cells . P26441 and TGF-beta synergize to induce P01282 mRNA expression and transcription through the P01282 CyRE . This synergy is dependent on the Smad , P35610 , and AP-1 sites , suggesting that these two independent cytokine pathways synergize through the cooperation of pathway-specific transcription factors binding to distinct sites within the P01282 CyRE . Opposite function of dopamine D1 and N-methyl-D-aspartate receptors in striatal cannabinoid-mediated signaling . It is well established that the cannabinoid and dopamine systems interact at various levels to regulate basal ganglia function . Although it is well known that acute administration of cannabinoids to mice can modify dopamine-dependent behaviors , the intraneuronal signaling pathways employed by these agents in the striatum are not well understood . Here we used knockout mouse models to examine the regulation of striatal extracellular-signal-regulated kinases 1 and 2 ( P27361 /2 ) signaling by behaviorally relevant doses of cannabinoids . This cellular pathway has been implicated as a central mediator of drug reward and synaptic plasticity . In C57BL/6J mice , acute administration of the cannabinoid agonists , (-)-11-hydroxydimethylheptyl-Δ8-tetrahydrocannabinol ( HU-210 ) and DB00470 ( Δ(9) -THC ) , promoted a dose- and time-dependent decrease in the phosphorylation of P27361 /2 in dorsal striatum . Co-administration of the P21554 cannabinoid receptor antagonist N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide(AM251) with HU-210 prevented P27361 /2 inactivation , indicating a requirement for activation of this receptor . In dopamine D1 receptor knockout animals treated with HU-210 , the magnitude of the HU-210-dependent decrease in striatal P27361 /2 signaling was greater than in wild-type controls . In contrast , HU-210 administration to N-methyl-D-aspartate receptor knockdown mice was ineffective at promoting striatal P27361 /2 inactivation . Genetic deletion of other potential P27361 /2 mediators , the dopamine D2 receptors or β-arrestin-1 or -2 , did not affect the HU-210-induced modulation of P27361 /2 signaling in the striatum . These results support the hypothesis that dopamine D1 receptors and N-methyl-D-aspartate receptors act in an opposite manner to regulate striatal P21554 cannabinoid receptor signal transduction . 7,12-Dimethylbenz[a]anthracene exposure induces the DNA repair response in neonatal rat ovaries . 7,12-Dimethylbenz[a]anthracene ( DMBA ) destroys ovarian follicles at all stages of development . This study investigated DMBA-induced DNA double strand break ( DSB ) formation with subsequent activation of the ovarian DNA repair response in models of pre-antral or pre-ovulatory follicle loss . Postnatal day ( P01160 ) 4 Fisher 344 ( F344 ) rat ovaries were cultured for 4 days followed by single exposures of vehicle control ( 1 % DB01093 ) or DMBA ( 12.5 nM or 75 nM ) and maintained in culture for 4 or 8 days . Alternately , PND4 F344 rat ovaries were exposed to 1 μM DMBA at the start of culture for 2 days . Total RNA or protein was isolated , followed by qPCR or Western blotting to quantify mRNA or protein level , respectively . γ P16104 and phosphorylated Q13315 were localized and quantified using immunofluorescence staining . DMBA exposure increased caspase 3 and γ P16104 protein . Additionally , DMBA ( 12.5 nM and 1 μM ) increased levels of mRNA encoding Atm , Xrcc6 , Brca1 and Rad51 . In contrast , Parp1 mRNA was decreased on d4 and increased on d8 of DMBA exposure , while P09874 protein increased after 8 days of DMBA exposure . Total Q13315 increased in a concentration-dependent temporal pattern ( 75 nM d4 ; 12.5 nM d8 ) , while pATM was localized in large primary and secondary follicles and increased after 8 days of 75 nM DMBA exposure compared to both control and 12.5 nM DMBA . These findings support that , despite some concentration effects , DMBA induces ovarian DNA damage and that DNA repair mechanisms are induced as a potential mechanism to prevent follicle loss . The cannabinoid DB00470 mediates inhibition of macrophage chemotaxis to RANTES/ P13501 : linkage to the CB2 receptor . The chemotactic response of murine peritoneal macrophages to RANTES/ P13501 was inhibited significantly following pretreatment with DB00470 ( THC ) , the major psychoactive component in marijuana . Significant inhibition of this chemokine directed migratory response was obtained also when the full cannabinoid agonist CP55940 was used . The CB2 receptor-selective ligand O-2137 exerted a robust inhibition of chemotaxis while the P21554 receptor-selective ligand ACEA had a minimal effect . The THC-mediated inhibition was reversed by the CB2 receptor-specific antagonist SR144528 but not by the P21554 receptor-specific antagonist SR141716A . In addition , THC treatment had a minimal effect on the chemotactic response of peritoneal macrophages from CB2 knockout mice . Collectively , these results suggest that cannabinoids act through the CB2 receptor to transdeactivate migratory responsiveness to RANTES/ P13501 . Furthermore , the results suggest that the CB2 receptor may be a constituent element of a network of G protein-coupled receptor signal transductional systems , inclusive of chemokine receptors , that act coordinately to modulate macrophage migration . Intestinal bacteria condition dendritic cells to promote IgA production . Immunoglobulin ( Ig ) A represents the predominant antibody isotype produced at the intestinal mucosa , where it plays an important role in limiting the penetration of commensal intestinal bacteria and opportunistic pathogens . We show in mice that Peyer 's Patch-derived dendritic cells ( PP-DC ) exhibit a specialized phenotype allowing the promotion of IgA production by B2 cells . This phenotype included increased expression of the retinaldehyde dehydrogenase 1 ( RALDH1 ) , inducible nitric oxide synthase ( P35228 ) , B cell activating factor of the tumor necrosis family ( Q9Y275 ) , a proliferation-inducing ligand ( APRIL ) , and receptors for the neuropeptide vasoactive intestinal peptide ( P01282 ) . The ability of PP-DC to promote anti- P25942 dependent IgA was partially dependent on retinoic acid ( RA ) and transforming growth factor ( TGF ) -beta , whilst Q9Y275 and APRIL signaling were not required . Signals delivered by Q9Y275 and APRIL were crucial for P25942 independent IgA production , although the contribution of B2 cells to this pathway was minimal . The unique ability of PP-DC to instruct naïve B cells to differentiate into IgA producing plasma cells was mainly imparted by the presence of intestinal commensal bacteria , and could be mimicked by the addition of LPS to the culture . These data indicate that exposure to pathogen-associated molecular patterns present on intestinal commensal bacteria condition DC to express a unique molecular footprint that in turn allows them to promote IgA production . The peripheral cannabinoid receptor 1 antagonist VD60 efficiently inhibits carbon tetrachloride-intoxicated hepatic fibrosis progression . This study investigated a peripheral selective CB₁ antagonist 3,4,22-3-demethoxycarbonyl-3-hydroxylmethyl-4-deacetyl-vindoline 3,4-thionocarbonate ( VD60 ) that efficiently inhibited hepatic fibrosis with lower psychological side effects . A competitive radiolabeled ligand binding experiment and DB02527 ( DB02527 ) response element-driven luciferase analysis were performed to evaluate the antagonistic activity of VD60 . Cell viability and collagen production were examined in the human hepatic stellate cell ( P19526 ) line LX-2 and primary cultured rat HSCs . The antifibrotic effects of VD60 were investigated in a CCl₄-induced liver fibrosis mouse model . The concentration of VD60 in the blood and the brain was determined by high-performance liquid chromatography-mass spectrum analysis . Furthermore , the potential underlying mechanisms of VD60 were investigated by Western blot . VD60 selectively competed with the radiolabeled P21554 agonist to bind to P21554 . VD60 antagonized P21554 agonist-induced Akt phosphorylation and increased the accumulation of intracellular DB02527 . VD60 strongly reduced the expression of α₂(I) pro-collagen mRNA and exerted potent antiproliferative effects on primary HSCs and LX-2 cells . The inhibition of reactive oxygen species production and phosphorylation of Akt , extracellular-signal-regulated kinase ( P29323 ) , and P84022 may explain the underlying mechanisms behind the antiproliferative effect of VD60 . Moreover , the in vivo antifibrotic activity of VD60 was confirmed in a CCl4-induced liver fibrosis mouse model . Most importantly , the concentration of VD60 in the peripheral blood was much higher than in the brain , suggesting that VD60 could act as a novel peripheral P21554 antagonist to efficiently inhibit hepatic fibrosis and could be used as a lead compound with low brain side effects in peripheral antifibrotic agents . Delta 9-tetrahydrocannabinol induces the apoptotic pathway in cultured cortical neurones via activation of the P21554 receptor . Delta 9-tetrahydrocannabinol , the principal psychoactive component of marijuana , exerts a variety of effects on the CNS , including impaired cognitive function and neurobehavioural deficits . The mechanisms underlying these neuronal responses to tetrahydrocannabinol are unclear but may involve alterations in neuronal viability . DB00470 has been shown to influence neuronal survival but the role of the cannabinoid receptors in the regulation of neuronal viability has not been fully clarified . In this study we demonstrate that tetrahydrocannabinol promotes the release of cytochrome c , activates caspase-3 , promotes cleavage of the DNA repair enzyme poly-ADP ribose polymerase and induces DNA fragmentation in cultured cortical neurones . These effects of tetrahydrocannabinol were completely abrogated by the CB(1) receptor antagonist AM-251 . The findings of this study demonstrate that tetrahydrocannabinol induces apoptosis in cortical neurones in a manner involving the P21554 subtype of cannabinoid receptor . Nicotinic alpha 7 receptors as a new target for treatment of cannabis abuse . Increasing use of cannabis makes the search for medications to reduce cannabis abuse extremely important . Here , we show that homomeric alpha7 nicotinic receptors are novel molecular entities that could be targeted in the development of new drugs for the treatment of cannabis dependence . In rats , systemic administration of the selective alpha7 nicotinic acetylcholine receptor antagonist methyllycaconitine ( MLA ) , but not the selective heteromeric non-alpha7 nicotinic acetylcholine receptor antagonist dihydrobetaerythroidine , ( 1 ) antagonized the discriminative effects of DB00470 ( THC ) , the main active ingredient in cannabis , ( 2 ) reduced intravenous self-administration of the synthetic cannabinoid P21554 receptor agonist WIN55,212-2 [ ( R ) -(+)-[2,3-dihydro-5-methyl-3[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-naphthalenyl)methanone , mesylate salt ] , and ( 3 ) decreased THC-induced dopamine elevations in the shell of the nucleus accumbens . Altogether , our results indicate that blockade of alpha7 nicotinic receptors reverses abuse-related behavioral and neurochemical effects of cannabinoids . Importantly , MLA reversed the effects of cannabinoids at doses that did not produce depressant or toxic effects , further pointing to alpha7 nicotinic antagonists as potentially useful agents in the treatment of cannabis abuse in humans . [ Cannabis use disorder and treatment of dependence ] . Cannabis , known as marijuana , has been used illicit drug by young people in the world . In our country , the number of user for cannabis is recently increased gradually . It has been suggested that regular use of cannabis might induce several adverse effects such as dependence syndrome , because DB00470 (THC) , a primary psychoactive component of cannabis , stimulates brain-reward areas through the activation of cannabinoid( P21554 ) receptor and induce drug-seeking behavior . Therefore , it is necessary to investigate and establish the medications for cannabis dependence . In fact , controlled laboratory studies and small open-label clinical studies have shown that several candidates of medications for cannabinoid dependence are identified . Further investigation in controlled clinical trials may produce the therapeutic benefit for treatment about cannabis-related problems . Reinforcing and neurochemical effects of cannabinoid P21554 receptor agonists , but not cocaine , are altered by an adenosine A2A receptor antagonist . Several recent studies suggest functional and molecular interactions between striatal adenosine A(2A) and cannabinoid CB(1) receptors . Here , we demonstrate that A(2A) receptors selectively modulate reinforcing effects of cannabinoids . We studied effects of A(2A) receptor blockade on the reinforcing effects of DB00470 ( THC ) and the endogenous CB(1) receptor ligand anandamide under a fixed-ratio schedule of intravenous drug injection in squirrel monkeys . A low dose of the selective adenosine A(2A) receptor antagonist MSX-3 ( 1 mg/kg ) caused downward shifts of THC and anandamide dose-response curves . In contrast , a higher dose of MSX-3 ( 3 mg/kg ) shifted THC and anandamide dose-response curves to the left . MSX-3 did not modify cocaine or food pellet self-administration . Also , MSX-3 neither promoted reinstatement of extinguished drug-seeking behavior nor altered reinstatement of drug-seeking behavior by non-contingent priming injections of THC . Finally , using in vivo microdialysis in freely-moving rats , a behaviorally active dose of MSX-3 significantly counteracted THC-induced , but not cocaine-induced , increases in extracellular dopamine levels in the nucleus accumbens shell . The significant and selective results obtained with the lower dose of MSX-3 suggest that adenosine A(2A) antagonists acting preferentially at presynaptic A(2A) receptors might selectively reduce reinforcing effects of cannabinoids that lead to their abuse . However , the appearance of potentiating rather than suppressing effects on cannabinoid reinforcement at the higher dose of MSX-3 would likely preclude the use of such a compound as a medication for cannabis abuse . DB00640 A(2A) antagonists with more selectivity for presynaptic versus postsynaptic receptors could be potential medications for treatment of cannabis abuse . Effects of DB00470 on human immune function and host defense . This review examines evidence that delta(9)-tetrahydrocannabinol ( THC ) can regulate and suppress human immune responses . Leukocytes express both cannabinoid receptor type 1 ( P21554 ) and cannabinoid receptor type 2 ( CB2 ) , and levels of mRNA encoding for them are increased in peripheral blood leukocytes obtained from marijuana smokers , suggesting cannabinoid receptor activation in vivo . Exposure of human T-cells to THC suppresses their proliferation , inhibits the release of interferon-gamma , and skews the balance of T-helper cytokines towards a type 2 response . The majority of these effects are CB2 receptor-dependent . Consistent with an impact of THC on cell-mediated immunity , alveolar macrophages ( AMs ) recovered from the lungs of marijuana smokers are suppressed in their ability to release pro-inflammatory cytokines and nitric oxide ( NO ) , and kill bacteria . Macrophage function is restored by treatment with interferon-gamma , a type 1 cytokine . Habitual exposure to THC appears capable of impacting on human cell-mediated immunity and host defense . Gender-dependent behavioral and biochemical effects of adolescent DB00470 in adult maternally deprived rats . Preclinical data support the long-term adverse effects on cognition , emotionality , and psychotic-like behaviors of adolescent exposure to natural and synthetic cannabinoids . To investigate whether the long-lasting adverse effects induced by cannabinoids in adolescence are influenced by early-life stress , female and male rats were subjected to 24-h maternal deprivation at postnatal day ( P01160 ) 9 and treated with tetrahydrocannabinol ( THC ) during adolescence ( P01160 35-45 ) according to our previously reported protocol . At adulthood , rats were tested in the novel object recognition , social interaction , and forced swim tests , to evaluate possible alterations in recognition memory , social behavior , and coping strategy . Moreover , cannabinoid P21554 receptor density and functionality , as well as DB01221 and dopamine D1 and D2 receptor densities were measured through autoradiographic binding studies . In female maternally deprived rats , THC failed to impair recognition memory , counteracted aggressiveness induced by maternal deprivation , whereas no interaction was observed in the passive coping behavior . In males , the association of the two events increased passive coping response without affecting other behaviors . This behavioral picture was accompanied by gender-dependent and region-specific alterations in DB01221 , D1 and D2 receptors . In conclusion , this study demonstrates that adolescent THC exposure might have different behavioral outcomes in animals previously exposed to early-life stress compared with non-stressed controls . The interaction between the two events is not univocal , and different combinations may arise depending on the sex of the animals and the behavior considered . Alterations in DB01221 , D1 and D2 receptors might be involved in the behavioral responses induced by maternal deprivation and in their modulation by THC . Anti-inflammatory activity of topical THC in DNFB-mediated mouse allergic contact dermatitis independent of P21554 and CB2 receptors . BACKGROUND : ∆(9) - DB00470 ( THC ) , the active constituent of Cannabis sativa , exerts its biological effects in part through the G-protein-coupled P21554 and CB2 receptors , which were initially discovered in brain and spleen tissue , respectively . However , THC also has P21554 /2 receptor-independent effects . Because of its immune-inhibitory potential , THC and related cannabinoids are being considered for the treatment of inflammatory skin diseases . Here we investigated the mechanism of the anti-inflammatory activity of THC and the role of P21554 and CB2 receptors . METHODS : We evaluated the impact of topically applied THC on DNFB-mediated allergic contact dermatitis in wild-type and P21554 /2 receptor-deficient mice . We performed immunohistochemical analyses for infiltrating immune cells and studied the influence of THC on the interaction between T cells , keratinocytes and myeloid immune cells in vitro . RESULTS : Topical THC application effectively decreased contact allergic ear swelling and myeloid immune cell infiltration not only in wild-type but also in P21554 /2 receptor-deficient mice . We found that THC ( 1 ) inhibited the production of IFNγ by T cells , ( 2 ) decreased the production of P13500 and of IFNγ-induced P80075 and CXL10 by epidermal keratinocytes and ( 3 ) thereby limited the recruitment of myeloid immune cells in vitro in a P21554 /2 receptor-independent manner . CONCLUSIONS : Topically applied THC can effectively attenuate contact allergic inflammation by decreasing keratinocyte-derived pro-inflammatory mediators that orchestrate myeloid immune cell infiltration independent of P21554 /2 receptors . This has important implications for the future development of strategies to harness cannabinoids for the treatment of inflammatory skin diseases . (+)- DB09061 analogues which bind cannabinoid receptors but exert peripheral activity only . Delta9- DB00470 ( Delta9-THC ) and (-)-cannabidiol are major constituents of the Cannabis sativa plant with different pharmacological profiles : (-)-Delta9-tetrahydrocannabinol , but not (-)-cannabidiol , activates cannabinoid P21554 and CB2 receptors and induces psychoactive and peripheral effects . We have tested a series of (+)-cannabidiol derivatives , namely , (+)-cannabidiol- Q03001 ( Q03001 -1,1-dimethylheptyl- ) , (+)-7-OH-cannabidiol- Q03001 , (+)-7-OH- cannabidiol , (+)-7-COOH- cannabidiol and (+)-7-COOH-cannabidiol- Q03001 , for central and peripheral ( intestinal , antiinflammatory and peripheral pain ) effects in mice . Although all (+)-cannabidiols bind to cannabinoid P21554 and CB2 receptors , only (+)-7-OH-cannabidiol- Q03001 was centrally active , while all (+)-cannabidiol analogues completely arrested defecation . The effects of (+)-cannabidiol- Q03001 and (+)-7-OH-cannabidiol- Q03001 were partially antagonized by the cannabinoid P21554 receptor antagonist N-(piperidiny-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide ( SR141716 ) , but not by the cannabinoid CB2 receptor antagonist N- [ -(1S)-endo-1,3,3-trimethil bicyclo [ 2.2.1 ] heptan-2-yl-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide ( SR144528 ) , and had no effect on P21554 (-/-) receptor knockout mice . (+)- DB09061 - Q03001 inhibited the peripheral pain response and arachidonic-acid-induced inflammation of the ear . We conclude that centrally inactive (+)-cannabidiol analogues should be further developed as antidiarrheal , antiinflammatory and analgesic drugs for gastrointestinal and other peripheral conditions . Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . Self-administration of cannabinoids by experimental animals and human marijuana smokers . Drug self-administration behavior has been one of the most direct and productive approaches for studying the reinforcing effects of psychoactive drugs , which are critical in determining their abuse potential . Cannabinoids , which are usually abused by humans in the form of marijuana , have become the most frequently abused illicit class of drugs in the United States . The early elucidation of the structure and stereochemistry of DB00470 ( THC ) in 1964 , which is now recognized as the principal psychoactive ingredient in marijuana , activated cannabinoid research worldwide . This review examines advances in research on cannabinoid self-administration behavior by humans and laboratory animals . There have been numerous laboratory demonstrations of the reinforcing effects of cannabinoids in human subjects , but reliable self-administration of cannabinoids by laboratory animals has only recently been demonstrated . It has now been shown that strong and persistent self-administration behavior can be maintained in experimentally and drug-naïve squirrel monkeys by doses of THC comparable to those in marijuana smoke inhaled by humans . Furthermore , reinforcing effects of some synthetic P21554 cannabinoid agonists have been recently reported using intravenous and intracerebroventricular self-administration procedures in rats and mice . These findings support previous conclusions that THC has a pronounced abuse liability comparable to other drugs of abuse under certain experimental conditions . Self-administration of THC by squirrel monkeys provides the most reliable animal model for human marijuana abuse available to date . This animal model now makes it possible to study the relative abuse liability of other natural and synthetic cannabinoids and to preclinically assess new therapeutic strategies for the treatment or prevention of marijuana abuse in humans .

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Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Induction of autophagy is an early response to gefitinib and a potential therapeutic target in breast cancer . Gefitinib ( DB00317 (®) , ZD1839 ) is a small molecule inhibitor of the epidermal growth factor receptor ( P00533 ) tyrosine kinase . We report on an early cellular response to gefitinib that involves induction of functional autophagic flux in phenotypically diverse breast cancer cells that were sensitive ( BT474 and SKBR3 ) or insensitive ( MCF7-GFPLC3 and JIMT-1 ) to gefitinib . Our data show that elevation of autophagy in gefitinib-treated breast cancer cells correlated with downregulation of AKT and P27361 /2 signaling early in the course of treatment . Inhibition of autophagosome formation by BECLIN-1 or O95352 siRNA in combination with gefitinib reduced the abundance of autophagic organelles and sensitized SKBR3 but not MCF7-GFPLC3 cells to cell death . However , inhibition of the late stage of gefitinib-induced autophagy with hydroxychloroquine ( HCQ ) or bafilomycin A1 significantly increased ( p < 0.05 ) cell death in gefitinib-sensitive SKBR3 and BT474 cells , as well as in gefitinib-insensitive JIMT-1 and MCF7-GFPLC3 cells , relative to the effects observed with the respective single agents . Treatment with the combination of gefitinib and HCQ was more effective ( p < 0.05 ) in delaying tumor growth than either monotherapy ( p > 0.05 ) , when compared to vehicle-treated controls . Our results also show that elevated autophagosome content following short-term treatment with gefitinib is a reversible response that ceases upon removal of the drug . In aggregate , these data demonstrate that elevated autophagic flux is an early response to gefitinib and that targeting P00533 and autophagy should be considered when developing new therapeutic strategies for P00533 expressing breast cancers . Lactobacillus protects the integrity of intestinal epithelial barrier damaged by pathogenic bacteria . Pathogens invade intestinal mucosal barrier through phagocytosis of antigen presenting cells ( dendritic cell , microfold cells ) , or through the invasion into the intestinal epithelial directly . Some pathogens could damage the cell junction between epithelial cells and use the paracellular pathway as an entrance to invade . Moreover , some Lactobacillus could inhibit the adhesion of the pathogens and protect the integrity of the cell junction and mucosal barrier . This research focused on the potential therapeutic effect of Lactobacillus fructosus ( L. fructosus ) P06681 to attenuate ETEC K88 or S. typhimurium SL1344 induced changes to mucosal barrier . The results demonstrated that treatment of polarized Caco-2 cells with L. fructosus P06681 reduced the permeation of dextran , and expression of P10145 , p- P29323 , and p-JNK when cells were infected with pathogenic bacteria . The findings indicated that L. fructosus P06681 exerted a protective effect against the damage to the integrity of Caco-2 cells by ETEC or S. typhimurium infection . HIV protease substrate conformation : modulation by cyclophilin A . P62937 ( CyPA ) , a cytosolic peptidyl-prolyl trans-cis isomerase can accelerate the trans-cis isomerization of Xxx-Pro peptide bonds . One- and two-dimensional 1H-NMR spectroscopy were used to determine that the heptapeptide DB00133 -Gln- DB00174 - DB00135 -Pro- DB00167 - DB00161 , a model peptide of an HIV-1 protease cleavage site in the gag polyprotein of HIV-1 , is a substrate for CyPA . Experiments revealed a slow exchange about the DB00135 -Pro peptide bond with 30 +/- 5 % in the cis conformation ( pH 1-9 ) . While the interconversion rate is too slow to measure by kinetic NMR methods in the absence of CyPA , these methods , saturation transfer and NOE experiments , established that CyPA enhanced the rate of trans-cis interconversion , a process inhibited by cyclosporin A ( DB00091 ) . With a substrate:CyPA ratio of 40:1 , an interconversion rate of 2.5 s(-1) at 25 degrees C was observed . P62937 facilitates translocation of the Clostridium botulinum P06681 toxin across membranes of acidified endosomes into the cytosol of mammalian cells . The binary Clostridium botulinum P06681 toxin consists of the binding/translocation component C2IIa and the separate enzyme component C2I , which mono-ADP-ribosylates actin in eukaryotic cells . Pore formation of C2IIa in early endosomal membranes facilitates translocation of unfolded C2I into the cytosol . We discovered earlier that translocation of C2I depends on the activity of the host cell chaperone heat shock protein Hsp90 . Here , we demonstrate that cyclosporin A , which inhibits the peptidyl-prolyl cis/trans isomerase activity of cyclophilins , inhibited intoxication of cells with P06681 toxin and prevented uptake of C2I into the cytosol . DB00091 blocked the pH-dependent translocation of C2I activity across membranes of intact cells and of partially purified early endosomes . In vitro , the addition of cytosol to P06681 toxin-loaded endosomes induced translocation of C2I activity into the cytosol , which was prevented by pretreatment of the cytosol with an antibody against cyclophilin A . Pull-down experiments with lysates from P06681 toxin-treated cells revealed specific binding of cyclophilin A to the N-terminal domain of C2I . In conclusion , our results suggest an essential role of cyclophilin A for translocation of C2I across endosomal membranes during the uptake of P06681 toxin into mammalian cells . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Differential expression of cyclophilin isoforms during keratinocyte differentiation . P62937 , the major intracellular binding protein for the immunosuppressive drug cyclosporin A ( DB00091 ) , was studied in human keratinocytes during differentiation both in vivo and in vitro . Analysis of cyclophilin by gel-filtration radiobinding-assay with tritiated DB00091 showed one specific radioactive peak at 17 kDa . By this technique , the levels of cyclophilin ( mean 55.23 +/- 8.43 pmol/mg protein ) did not significantly differ during keratinocyte differentiation . When the protein extracts from calcium-induced differentiating keratinocytes and normal human skin were analysed by PAGE radiobinding-assay , two specific radioactive DB00091 -binding peaks were detected . The major peak ( RF 0.13 ) was expressed in all samples ( mean 47.32 +/- 17.53 pmol/mg protein ) whereas the minor peak ( RF 0.23 ) was dramatically decreased about 6-fold in abnormally differentiated skin ( psoriasis ) as well as in non-differentiated keratinocytes . At least six [3H] DB00091 -binding isoforms with pI values ranging from 5.58 to 7.75 were detected by isoelectrofocusing autoradio-blotting-assay in normal human skin ; three of them immunoreacted with antibodies to cyclophilin . These results demonstrated the presence of several cyclophilin isoforms in human epidermal cells and an expression which correlated with the differentiation of human keratinocytes both in vivo and in vitro . SERCA2b activity is regulated by cyclophilins in human platelets . OBJECTIVE : The role of cyclophilins ( chaperones that are widely expressed in different cell types , including human platelets ) was explored in sarcoendoplasmic calcium ( Ca(2+) ) adenosine triphosphatase ( SERCA ) activity . METHODS AND RESULTS : Cyclophilin inhibition by cyclosporin A ( DB00091 ) evoked a time- and concentration-dependent reduction of Ca(2+) uptake by SERCA2b . However , other Ca(2+)-adenosine triphosphatases expressed in platelets , such as Q93084 and plasma membrane Ca(2+) adenosine triphophatase , remained unaltered after DB00091 treatment . Cypermethrin , a non- DB00091 -related calcineurin inhibitor , did not alter SERCA2b activity . Furthermore , SERCA2b was affected by other DB00091 analogues , which do not interfere with calcineurin , such as PKF-211-811-NX5 ( NIM811 ) and sanglifehrin A . Inhibition of the immunophilin family members using FK506 ( tacrolimus ) did not alter SERCA2b ability to sequester Ca(2+) into the dense tubular system . Coimmunoprecipitation experiments confirmed that cyclophilin A associates with SERCA2b and stromal interaction molecule-1 in resting platelets . This interaction is attenuated by the physiological agonist thrombin but enhanced by treatment with DB00091 or sanglifehrin A . CONCLUSIONS : P62937 is a regulator of SERCA2b in human platelets . Biophysical and pharmacological characterization of hypotonically activated chloride currents in cortical astrocytes . Rat cortical astrocytes regulate their cell volume in response to hypotonic challenge . This regulation is believed to depend largely on the release of chloride or organic osmolytes through anion channels . Using whole-cell recordings , we identified weakly outwardly rectifying chloride currents that could be activated in response to hypotonic challenge . These currents exhibited the following permeability sequence upon replacement of chloride in the bathing solution with various anions : I- > NO3- > Cl- > Gluc- > or = MeS- > Ise- . Interestingly , extracellular I- , albeit showing the greatest permeability , blocked the currents with an IC50 of approximately 50 mM . Currents were almost completely inhibited by 123 microM P16860 and partially inhibited by 200 microM niflumic acid or 200 microM DIDS . Additionally , the total number of Cl- ions effluxed through the hypotonically activated channels was markedly similar to the total solute efflux during volume regulation . We therefore propose the hypotonically activated chloride channel as a major contributor to volume regulation of astrocytes . To examine potential candidate chloride channel genes expressed by astrocytes , we employed RT-PCR to demonstrate the presence of transcripts for P51788 , 3 , 4 , 5 , and 7 , as well as for P21796 and P13569 in cultured astrocytes . Moreover , we performed immunostaining with antibodies against each of these channels and showed the strongest expression of P51788 and P51790 , strong expression of P51795 and P21796 , weak expression of P51798 and very weak expression of P51793 and P13569 . Intriguingly , although we found at least seven Cl- channel proteins from three different gene families in astrocytes , none appeared to be active in resting cells . Antithrombin III concentrate in the acute phase of thermal injury . BACKGROUND : Thermal injury disrupts homeostasis by inducing subclinical disseminated intravascular coagulation , fibrinolysis. and an acquired deficiency of Antithrombin III ( P01008 ) , a natural anticoagulant . As a result , thermally injured patients have a high incidence of hypercoagulability and thrombosis . OBJECTIVE : P01008 ( Human ) concentrate was given to a thermally injured patient to evaluate safety , and dosage requirements in this setting . DESIGN : The patient was a 40 yr old male with a 68 % total burn surface area , right femoral comminuted fracture , and P01031 - P13671 subluxation sustained in a vehicular crash . He received nine infusions of AT III ( H ) concentrate ( 100-50 u/kg ) within the first four days of injury . RESULT : The P01008 plasma level increased from 45 % on admission ( normal = 100+/-20 % ) to 120+/-25 % in the next four days . During the 64 day hospitalization , there were 11 grafting procedures with an estimated blood loss ( EBL ) /procedure : 1140 cc ; and EBL/grafted surface area ratio : 0.6 cc cm2 . The average time to healing of the meshed autograft was 6.4 days . CONCLUSION : P01008 ( H ) concentrate can be safely utilized in the acute phase of thermal injury : no excessive bleeding or prolongation of wound healing was documented . Interferon-gamma-induced dephosphorylation of P40763 and apoptosis are dependent on the P42345 pathway . Interferon-gamma ( P01579 ) exhibits diverse biological activities , including control of cell growth and tumor suppression . Here , we report that the treatment of M12 cells , a human metastatic prostate cancer cell line , with P01579 , resulted in marked inhibition of cell proliferation and induced apoptosis . These effects were not seen with either IFN-alpha or IFN-beta . M12 cells , like many other human cancer cells , contain constitutively activated signal transducer and activator of transcription 3 ( P40763 ) . The basal levels of both Akt and P27361 /2 phosphorylation are also markedly elevated in M12 cells . Strikingly , P01579 -induced apoptosis and growth inhibition of M12 cells were associated with persistent suppression of the constitutive tyrosine-phosphorylated P40763 ( pY- P40763 ) . The P01579 -induced dephosphorylation of pY- P40763 , however , was inhibited when the P42345 pathway was specifically blocked by rapamycin . Inhibition of PI-3K with low-dose LY294002 , or MAPK with PD98059 also suppressed the P42345 / P08133 S6k pathway , and correlated with the blockage of P01579 -induced dephosphorylation of pY- P40763 . Simultaneously , treatment with LY294002 , PD98059 , or rapamycin abolished P01579 -induced apoptosis in M12 cells . The inhibition of the P42345 pathway , however , did not affect P01579 -induced activation of P42224 pathway , and suppression of P42224 expression by siRNA had no effect on P01579 -induced dephosphorylation of pY- P40763 . Taken together , these results demonstrate that an intact P42345 pathway is critical for P01579 -induced suppression of pY- P40763 and apoptosis . Our study thus provides novel insights into the contributions of signaling pathways other than the classical JAK/ P42224 pathway in the anti-proliferative , proapoptotic actions of P01579 . P62937 is required for Q9C035 {alpha}-mediated resistance to HIV-1 in Old World monkey cells . The peptidyl-prolyl isomerase cyclophilin A ( CypA ) embraces an exposed , proline-rich loop on HIV-1 capsid ( CA ) and renders reverse transcription complexes resistant to an antiviral activity in human cells . A CypA fusion with Q9C035 that is unique to New World owl monkeys also targets HIV-1 CA , but this interaction potently inhibits infection . A similar block to HIV-1 infection in Old World monkeys is attributable to the alpha isoform of the Q9C035 orthologue in these species . To determine whether HIV-1 restriction by Old World monkey TRIM5alpha is modulated by the CA-CypA interaction , RNA interference was used to disrupt CypA in cells from African green monkeys and rhesus macaques . HIV-1 infectivity increased in response to CypA knock-down to the same extent that it increased in response to Q9C035 knock-down . CypA knock-down eliminated the HIV-1 stimulatory effect of cyclosporin A ( DB00091 ) , a competitive inhibitor of the CypA-CA interaction , or of CA mutants that block binding to CypA but caused no change in titer of retroviruses that do n't interact with CypA . Simultaneous knock-down of both CypA and Q9C035 caused minimal additional increase in titer , suggesting that CypA inhibits HIV-1 replication in these cells because it is required for CA recognition by TRIM5alpha . Finally , DB00091 increased HIV-1 titer in otherwise nonrestrictive feline cells but only after these cells were transduced with Old World monkey TRIM5alpha . Thus , CypA is required for HIV-1 restriction by Old World monkey orthologues of TRIM5alpha . Two cyclophilin A homologs with shared and distinct functions important for growth and virulence of Cryptococcus neoformans . P62937 is the target of the immunosuppressant cyclosporin A ( DB00091 ) and is encoded by a single unique gene conserved from yeast to humans . In the pathogenic fungus Cryptococcus neoformans , two homologous linked genes , P15085 and P48052 , were found to encode two conserved cyclophilin A proteins . In contrast to Saccharomyces cerevisiae , in which cyclophilin A mutations confer DB00091 resistance but few other phenotypes , cyclophilin A mutations conferred dramatic phenotypes in C. neoformans . The Cpa1 and Cpa2 cyclophilin A proteins play a shared role in cell growth , mating , virulence and DB00091 toxicity . The Cpa1 and Cpa2 proteins also have divergent functions . cpa1 mutants are inviable at 39 degrees C and attenuated for virulence , whereas cpa2 mutants are viable at 39 degrees C and fully virulent . cpa1 cpa2 double mutants exhibited synthetic defects in growth and virulence . P62937 active site mutants restored growth of cpa1 cpa2 mutants at ambient but not at higher temperatures , suggesting that the prolyl isomerase activity of cyclophilin A has an in vivo function . The study of genetic polymorphisms related to serotonin in Alzheimer 's disease : a new perspective in a heterogenic disorder . Genetic and environmental factors have been implicated in the development of Alzheimer 's disease ( AD ) , the most common form of dementia in the elderly . Mutations in 3 genes mapped on chromosomes 21 , 14 and 1 are related to the rare early onset forms of AD while the epsilon 4 allele of the apolipoprotein E ( P02649 ) gene ( on chromosome 19 ) is the major susceptibility locus for the most common late onset AD ( LOAD ) . Serotonin ( 5-hydroxytryptamine or 5-HT ) is a key neurotransmitter implicated in the control of mood , sleep , appetite and a variety of traits and behaviors . Recently , a polymorphism in the transcriptional control region upstream of the 5-HT transporter ( 5-HTT ) gene has been studied in several psychiatric diseases and personality traits . It has been demonstrated that the short variant(s) of this 5-HTT gene-linked polymorphic region ( 5-HTTLPR ) is associated with a different transcriptional efficiency of the 5-HTT gene promoter resulting in decreased 5-HTT expression and 5-HT uptake in lymphocytes . An increased frequency of this 5-HTTLPR short variant polymorphism in LOAD was recently reported . In addition , another common polymorphic variation in the 5- Q13049 and P28335 serotonin receptor genes previously analyzed in schizophrenic patients was associated with auditory and visual hallucinations in AD . These observations suggest that the involvement of the serotonin pathway might provide an explanation for some aspects of the affective symptoms commonly observed in AD patients . In summary , research on genetic polymorphisms related to AD and involved in receptors , transporter proteins and the enzymatic machinery of serotonin might enhance our understanding of this devastating neurodegenerative disorder . A cyclophilin A inducible expressed in gonad of zhikong scallop Chlamys farreri . P62937 ( CypA ) , a receptor for the immunosuppressive agent cyclosporin A ( DB00091 ) , is a cis-trans peptidyl-prolyl isomerase ( PPIase ) which accelerates the cis-trans isomerization of prolyl-peptide bonds , interacts with a variety of proteins and therefore regulates their activities . One CypA ( designated CfCypA ) cDNA was cloned from Chlamys farreri by expressed sequence tag ( EST ) and rapid amplification of cDNA ends ( RACE ) techniques . The full-length cDNA of CfCypA consisted of 1,248 nucleotides with a canonical polyadenylation signal sequence AATAAA , a poly ( A ) tail , and an open reading frame ( ORF ) of 495 nucleotides encoding a polypeptide of 164 amino acids . The deduced amino acid sequence shared high similarity with CypA from the other species , indicating that CfCypA should be a new member of the CypA family . Quantitative real-time ( RT ) PCR was employed to assess the mRNA expression of CfCypA in various tissues and its temporal expression in haemocytes and gonad of scallops challenged with Vibrio anguillarum . The mRNA transcripts of CfCypA could be detected in all the examined tissues with highest expression level in gonad . After bacterial challenge , the expression level of CfCypA was almost unchanged in haemocytes , but up-regulated in gonad and increased to the peak ( 22.59-fold ; P < 0.05 ) at 4 h post-injection , and then dropped to the original level at 8 h post-injection . These results indicated that CfCypA was constitutive expressed in haemocytes , but could be induced in gonad , and perhaps played a critical role in response to the bacterial challenge in gonad . Essential role for retinoic acid in the promotion of P01730 (+) T cell effector responses via retinoic acid receptor alpha . DB00162 and its metabolite , retinoic acid ( RA ) are implicated in the regulation of immune homeostasis via the peripheral induction of regulatory T cells . Here we showed RA was also required to elicit proinflammatory P01730 (+) helper T cell responses to infection and mucosal vaccination . P10276 ( RARα ) was the critical mediator of these effects . Antagonism of RAR signaling and deficiency in RARα ( Rara(-/-) ) resulted in a cell-autonomous P01730 (+) T cell activation defect , which impaired intermediate signaling events , including calcium mobilization . Altogether , these findings reveal a fundamental role for the RA-RARα axis in the development of both regulatory and inflammatory arms of adaptive immunity and establish nutritional status as a broad regulator of adaptive T cell responses . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . P14735 binds to the nonglycosylated precursor of varicella-zoster virus gE protein found in the endoplasmic reticulum . P01308 degradation enzyme ( P14735 ) is a 110-kDa zinc metalloprotease found in the cytosol of all cells . P14735 degrades insulin and a variety of small proteins including amyloid-beta . Recently , P14735 has been proposed as the receptor for varicella-zoster virus ( VZV ) attachment . During our reassessment , some of the original studies were repeated and expanded in scope . We first confirmed that P14735 antibody reduced VZV spread . For additional controls , we repeated the same experiments with herpes simplex virus ( HSV ) -infected cells as well as uninfected cells . There was a visible reduction in HSV spread but less than seen in the VZV system . Of greater importance , P14735 antibody also inhibited the growth of uninfected cells . Second , we repeated the coprecipitation assays . We confirmed that antibodies to VZV gE ( open reading frame 68 ) coprecipitated P14735 and that anti- P14735 antibody coprecipitated gE . However , the detected gE protein was not the mature 98-kDa form ; rather , it was a precursor 73-kDa gE form found in the endoplasmic reticulum . Additional control experiments included VZV-infected cell cultures treated with tunicamycin to block gE glycosylation in the endoplasmic reticulum ; again , the anti- P14735 antibody coprecipitated a 73-kDa gE product . Finally , Orbitrap mass spectrometry analysis of a chromatographically purified gE sample revealed four cellular proteins associated with the unfolded protein response : P11021 ( P11021 ) , P11142 , P10809 , and P62937 ( peptidyl-propyl cis-trans isomerase ) . We conclude that P14735 protease binds to the 73-kDa gE precursor and that this event occurs in the cytosol but not as a receptor/ligand interaction . [ The mechanisms of the resistance to molecular targeting agents ] . Increasing knowledge of the mechanism of the initiation and progression of various cancers is the catalyst for developing new anticancer therapeutics that target specific molecules expressed in cancer cells . STI571 ( imatinib mesylate ) is an example of the successful development of a rationally designed and targeted agent . Its target is the constitutively active tyrosine kinase , P11274 - P00519 in chronic myelogenous leukemia ( CML ) . Clinical studies with STI571 in CML demonstrated that many patients with advanced stage disease respond initially but then relapse . Drug resistance is associated with the reactivation of P11274 - P00519 signal transduction . Another targeted protein-tyrosine kinase inhibitor that was approved for clinical use is ZD1839 ( DB00317 ) . ZD1839 is an orally active and selective inhibitor for epidermal growth factor receptor ( P00533 ) tyrosine kinase . P04626 is overexpressed in 25-30 % of breast cancers and associated with shorter time to relapse and lower survival rate . Specific targeting of these cancers can be accomplished with Herceptin directed against the extracellular domain of the P04626 protein . However , even in the selected group of patients with high levels of P04626 , the response to Herceptin is limited in magnitude and duration . The mechanisms of the resistance to these targeted agents were reviewed . Cyclophilin interactions with incoming human immunodeficiency virus type 1 capsids with opposing effects on infectivity in human cells . P62937 ( CypA ) is a peptidyl-prolyl isomerase that binds to the capsid protein ( CA ) of human immunodeficiency virus type 1 ( HIV-1 ) and by doing so facilitates HIV-1 replication . Although CypA is incorporated into HIV-1 virions by virtue of CypA-Gag interactions that occur during virion assembly , in this study we show that the CypA-CA interaction that occurs following the entry of the viral capsid into target cells is the major determinant of CypA 's effects on HIV-1 replication . Specifically , by using normal and CypA-deficient Jurkat cells , we demonstrate that the presence of CypA in the target and not the virus-producing cell enhances HIV-1 infectivity . Moreover , disruption of the CypA-CA interaction with cyclosporine A ( DB00091 ) inhibits HIV-1 infectivity only if the target cell expresses CypA . The effect of DB00091 on HIV-1 infection of human cells varies according to which particular cell line is used as a target , and CA mutations that confer DB00091 resistance and dependence exert their effects only if target cells , and not if virus-producing cells , are treated with DB00091 . The differential effects of DB00091 on HIV-1 infection in different human cells appear not to be caused by polymorphisms in the recently described retrovirus restriction factor TRIM5alpha . We speculate that CypA and/or CypA-related proteins affect the fate of incoming HIV-1 capsid either directly or by modulating interactions with unidentified host cell factors . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . The proteomic study of sodium butyrate antiproliferative/cytodifferentiation effects on K562 cells . Employing methods of cell biology and proteome analysis tools , we examined effects of an inhibitor of histone deacetylases , sodium butyrate ( SB ) , on the proliferation/differentiation characteristics of chronic myelogenous leukemia ( CML ) -derived cells K562 . SB suppressed proliferation of K562 cells by inducing cell cycle arrest in P55008 phase , which was followed by their transition to G0 phase ( decrease of Ki-67 antigen-positive cells ) and erythroid differentiation ( increased glycophorin A expression and synthesis of hemoglobins ) . Neither terminal apoptosis ( low counts of TUNEL-positive cells ) nor necrosis ( moderate counts of propidium iodide-positive cells ) occurred . Importantly , SB attenuated protein expression of CML-related chimeric kinase P11274 - P00519 that is responsible for the deregulated proliferation of CML cells . The proteomic analysis ( 2-D electrophoresis combined with MALDI-TOF mass spectrometry and/or Western blotting ) revealed several proteins that were differentially expressed or their mobility was altered due to butyrate treatment , namely , HSP90 , HSP70 , p23 , cyclophilin A ( P62937 ) , prefoldin2 ( Q9UHV9 ) and alpha- , gamma- , epsilon-human globin chains . Perturbation of HSP90 multichaperone complex of which P11274 - P00519 is the client protein is presumably a cause of P11274 - P00519 suppression . Changes in other proteins with chaperonic functions , P62937 and Q9UHV9 , may reflect SB antiproliferative and cytodifferentiation effects . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Nephrotoxicity of cyclosporin A and FK506 : inhibition of calcineurin phosphatase . DB00091 ( DB00091 ; 50 mg/kg ) and Fujimycine ( FK506 ; 5 mg/kg ) , but not the related macrolide immunosuppressant rapamycin ( 5 mg/kg ) , caused a reduction of glomerular filtration rate , degenerative changes of proximal tubular epithelium , and hypertrophy of the juxtaglomerular apparatus in male Wistar rats when given for 10 days . The molecular mechanisms of DB00091 and FK506 toxicity were investigated . P62937 and FK506-binding protein , the main intracytoplasmic receptors for DB00091 and FK506 , respectively , were each detected in renal tissue extract . In the kidney , high levels of immunoreactive and enzymatically active calcineurin were found which were inhibited by the immunosuppressants DB00091 and FK506 , but not by rapamycin . Finally , specific immunophilin-drug-calcineurin complexes formed only in the presence of DB00091 and FK506 , but not rapamycin . These results suggest that the nephrotoxic effects of DB00091 and FK506 is likely mediated through binding to renal immunophilin and inhibiting calcineurin phosphatase . Proteomics-based identification of a group of apoptosis-related proteins and biomarkers in gastric cancer . Gastric cancer ( GC ) is the one of the most common types of cancer in Asia . To better understand the molecular mechanisms underlying GC , and to seek new markers of tumor progression , we used a proteomics strategy to analyze the protein expression patterns in matched pairs of GC tissue and normal gastric mucosa of 8 GC patients . Comparative proteomic analysis , using two-dimensional gel electrophoresis ( 2-DE ) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry ( MALDI-TOF-MS ) , revealed that 32 protein spots showed a > 2-fold difference in intensity between tumor and normal tissues . Twenty-six proteins were up-regulated and 6 proteins were down-regulated in tumor tissue compared to control . Western blot analysis confirmed differential expression for 9 proteins , including O95994 , P06733 , P50395 , P11021 , P14625 , P62937 , Q06830 , P60484 and P21796 . Immunohistochemical staining of a tissue microarray , derived from 145 GC patients , with antibodies for each of the 9 proteins demonstrated a significant association between the level of protein immunostaining and the clinical features of the disease in the donor . The identified proteins were functionally classified using bioinformatics methods , showing that the 9 proteins identified were related to P10415 , Q07812 , P04626 and P42574 proteins and involved in the process of apoptosis . These proteomic data provide potentially valuable insights into both the biology of GC and the identity of biomarkers for tumor progression . We propose P06733 , P11021 , P14625 , P62937 , Q06830 and P60484 as potential GC biomarkers . [ DB00091 inhibits the adhesion of neutrophil with ECV-304 induced by hypoxia/reoxygenation via ROS- P62937 - P27361 /2 pathway ] . To investigate the inhibition of cyclosporin A ( DB00091 ) on neutrophil adhesion to human umbilical vein endothelial cells ( HUVECs , ECV-304 ) induced by hypoxia/reoxygenation and further explore its mechanism , a 1 h hypoxia/4 h reoxygenation model was reproduced using ECV-304 . The adhesion rate of neutrophils to ECV-304 was determined by measuring the activity of endogenous hexosaminidase . The expression of endothelial cell adhesion molecules of P16581 and P05362 was measured by flow cytometry . The expression of cyclophilin A ( CyPA ) and the activation of P27361 /2 was compared among experimental groups by Western blot . The content of reactive oxygen species ( ROS ) was measured by Fenton reaction . After being stimulated with 1 h hypoxia/4 h reoxygenation , ECV-304 showed an enhanced neutrophil adhensiveness in association with an increased surface expression of P16581 and P05362 . In parallel , the content of ROS was also increased . These effects were significantly suppressed by the addition of DB00091 . Most importantly , the expression of CyPA was significantly increased following 1 h hypoxia/4 h reoxygenation , which was accompanied with an increased activation of P27361 /2 . Treatment with CyPA inhibitor DB00091 and CyPA antisense oligonucleotides significantly inhibited the activation of P27361 /2 and decreased the adhesion of neutrophils to ECV-304 . The specific P27361 /2 inhibitor PD98059 caused an inhibition of neutrophil adhesion to hypoxia/reoxygenation-stimulated ECV-304 . Our data confirm that DB00091 inhibits neutrophil adhesion to hypoxia/reoxygenation stimulated ECV-304 by a mechanism involving inhibition of the signal transduction of ROS , CyPA and P27361 /2 . In utero activation of fetal memory T cells alters host regulatory gene expression and affects HIV susceptibility . In utero priming to malaria antigens renders cord blood mononuclear cells ( CBMC ) more susceptible to productive HIV infection in vitro in the absence of exogenous stimulation . This provides a unique model to better understand mechanisms affecting lymphocyte susceptibility to HIV infection in vivo . Effector memory CD3(+) P01730 (+) T cells ( T(EM) ) were the exclusive initial targets of HIV with rapid spread to central memory cells . HIV susceptibility correlated with increased expression of CD25 and HLA-DR on T(EM) . Virus entered all samples equally , however gag/pol RNA was only detected in HIV susceptible samples , suggesting regulation of proviral gene transcription . Targeted analysis of human genes in memory T cells showed greater expression of P01579 , O95644 , P10914 , P01100 , and P62937 and decreased expression P25490 and Q12800 in HIV susceptible samples . Thus fetal priming to exogenous antigens enhances specific proviral gene transcription pathways in effector memory cells that may increase risk of vertical transmission of HIV . P62937 -deficient mice are resistant to immunosuppression by cyclosporine . DB00091 is an immunosuppressive drug that is widely used to prevent organ transplant rejection . Known intracellular ligands for cyclosporine include the cyclophilins , a large family of phylogenetically conserved proteins that potentially regulate protein folding in cells . Immunosuppression by cyclosporine is thought to result from the formation of a drug-cyclophilin complex that binds to and inhibits calcineurin , a serine/threonine phosphatase that is activated by TCR engagement . Amino acids within the cyclophilins that are critical for binding to cyclosporine have been identified . Most of these residues are highly conserved within the 15 mammalian cyclophilins , suggesting that many are potential targets for the drug . We examined the effects of cyclosporine on immune cells and mice lacking Ppia , the gene encoding the prototypical cyclophilin protein cyclophilin A . TCR-induced proliferation and signal transduction by Ppia(-/-) P01730 (+) T cells were resistant to cyclosporine , an effect that was attributable to diminished calcineurin inhibition . Immunosuppressive doses of cyclosporine failed to block the responses of Ppia(-/-) mice to allogeneic challenge . Rag2(-/-) mice reconstituted with Ppia(-/-) splenocytes were also cyclosporine resistant , indicating that this property is intrinsic to Ppia(-/-) immune cells . Thus , among multiple potential ligands , CypA is the primary mediator of immunosuppression by cyclosporine . P62937 and calcineurin functions investigated by gene inactivation , cyclosporin A inhibition and cDNA arrays approaches in the phytopathogenic fungus Botrytis cinerea . Calcineurin phosphatase and cyclophilin A are cellular components involved in fungal morphogenesis and virulence . Their roles were investigated in the phytopathogenic fungus Botrytis cinerea using gene inactivation , drug inhibition and cDNA macroarrays approaches . First , the BCP1 gene coding for cyclophilin A was identified and inactivated by homologous recombination . The bcp1Delta null mutant obtained was still able to develop infection structures but was altered in symptom development on bean and tomato leaves . Opposite to this , calcineurin inhibition using cyclosporin A ( DB00091 ) modified hyphal morphology and prevented infection structure formation . DB00091 drug pattern signature on macroarrays allowed the identification of 18 calcineurin-dependent ( CND ) genes among 2839 B. cinerea genes . Among the co-regulated CND genes , three were shown to be organized as a physical cluster that could be involved in secondary metabolism . The signature of BCP1 inactivation on macroarrays allowed the identification of only three BCP1 cyclophilin-dependent ( O75976 ) genes that were different from CND genes . Finally , no DB00091 drug pattern signature was observed in the bcp1Delta null mutant which provided a molecular target validation of the drug . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . P62937 , the major intracellular receptor for the immunosuppressant cyclosporin A , maps to chromosome 7p11.2-p13 : four pseudogenes map to chromosomes 3 , 10 , 14 , and 18 . P62937 ( CyP-A ) , the major intracellular receptor for the immunosuppressant cyclosporin A ( DB00091 ) , is a member of the immunophilin class of proteins , which all possess peptidyl-prolyl cis-trans isomerase activity and , therefore , are believed to be involved in protein folding and/or intracellular protein transport . The CyP-A protein is encoded by a single gene ; in addition , 15 pseudogenes have been identified . Recently , specific binding of CyP-A to the human immunodeficiency virus type 1 ( HIV-1 ) gag protein has been reported . Interestingly , this interaction can be inhibited by the immunosuppressant DB00091 and also by nonimmunosuppressive , CyP-A-binding DB00091 derivatives , which were also shown to exhibit potent anti-HIV-1 activity . Results thus indicate that CyP-A may have an essential function in HIV-1 replication . Using a panel of somatic rodent-human cell hybrids and PCR technology , we localized the coding cyclophilin A gene ( P62937 ) on chromosome 7 and four pseudogenes ( PPIP2 , PPIP3 , PPIP4 , and PPIP6 ) on chromosomes 14 , 10 , 18 , and 3 , respectively . Using chromosome 7 and chromosome 10 deletion hybrid panels , we were able to localize further the coding gene to the region 7p11.2-p13 , as confirmed by fluorescence in situ hybridization analysis , and one pseudogene ( PPIP3 ) to the region 10q11.2-q23 . This is the first report on the regional mapping of members of the CyP-A gene family . P62937 as a target of DB00515 chemosensitizers . Platinum-based chemotherapeutics are the mainstay of treatment of a range of tumors achieving high response rates but limited in the course of disease by appearance of drug resistance . Tumor cells respond with reduced uptake and increased intracellular inactivation of the drugs , as well as increased DNA repair and general resistance to chemotherapyinduced cell death . DB00515 is known to induce expression of cyclophilins , a group of proteins that have peptidyl-prolyl cis-trans isomerase ( PPIase ) and molecular chaperone activities , as stress response . P62937 ( CypA ) and other members of this family are inhibited by cyclosporin A ( DB00091 ) which sensitized diverse drug-resistant tumor cell lines in vitro to cisplatin . This effect of DB00091 was attributed to metabolic changes , inhibition of DNA repair , enhancement of apoptosis , altered intracellular signal transduction or increased production of reactive oxygen species ( ROS ) , although no definitive explanation was provided so far . Several clinical trials employing cisplatin/carboplatin in combination with DB00091 yielded unsatisfactory results . Since viral replication was found to be dependent on cyclophilins of the host cells , effective new inhibitors , different from DB00091 or with low or absent immunosuppressive activity , are in development or clinical trials . Sanglifehrins are more potent than DB00091 and proved to increase toxicity of cisplatin against hepatocellular cancer cells in vitro . These novel cyclophilin inhibitors may offer new opportunities to achieve reversal of resistance to platinumbased drugs in refractory patients . Responsive cancer patients may be enriched in clinical trials by an identification of the downstream targets of Cyps responsible for chemoresistance . The role of the central flexible region on the aggregation and conformational properties of human ataxin-3 . Aggregation of human ataxin-3 ( P01008 ) into amyloid fibrils is responsible for spinocerebellar ataxia type 3 . This protein consists of a folded N-terminal domain ( Josephin domain , residues 1-182 ) , a central flexible region ( residues 183-291 ) , a poly-glutamine sequence of variable length and a short C-terminal flexible region . Very little is known about the influence of the central flexible region on the conformational and aggregation properties of this protein . The present study aimed to investigate the specific role of this portion of the protein ( residues 183-291 ) . Accordingly , protein fragments 1-182 ( P01008 /182 ) and 1-291 ( P01008 /291 ) were produced and compared by thioflavin-T fluorescence , Fourier transform infrared spectroscopy , CD , intrinsic fluorescence and P19957 -MS . It is shown that the central flexible region enhances protein aggregation and can populate conformational states with different degrees of compactness . Both monomeric and dimeric partially-folded forms are identified for both protein fragments under denaturing conditions . Partially-folded monomers and dimers accumulate to a larger extent in P01008 /291 . These species represent good candidates for early intermediates of the aggregation process under the experimental conditions employed in the present study . Secreted cyclophilin A , a peptidylprolyl cis-trans isomerase , mediates matrix assembly of hensin , a protein implicated in epithelial differentiation . Q9UGM3 is a rabbit ortholog of Q9UGM3 , a multifunctional , multidomain protein implicated in the regulation of epithelial differentiation , innate immunity , and tumorigenesis . Q9UGM3 in the extracellular matrix ( Q13201 ) induced morphological changes characteristic of terminal differentiation in a clonal cell line ( clone C ) of rabbit kidney intercalated cells . Although hensin is secreted in monomeric and various oligomeric forms , only the polymerized Q13201 form is able to induce these phenotypic changes . Here we report that hensin secretion and matrix assembly were inhibited by the peptidylprolyl cis-trans isomerase ( PPIase ) inhibitors cyclosporin A ( DB00091 ) and a derivative of cyclosporin A with modifications in the d- DB00133 side chain ( Cs9 ) but not by the calcineurin pathway inhibitor FK506 . PPIase inhibition led to failure of hensin polymerization in the medium and Q13201 , plus the loss of apical cytoskeleton , apical microvilli , and the columnar epithelial shape of clone C cells . P62937 was produced and secreted into the media to a much greater extent than cyclophilins B and C . Our results also identified the direct DB00091 -sensitive interaction of cyclophilin A with hensin , suggesting that cyclophilin A is the PPIase that mediates the polymerization and matrix assembly of hensin . These results are significant because this is the first time a direct role of peptidylprolyl cis-trans isomerase activity has been implicated in the process of epithelial differentiation . Cyclophilin inhibitors as a novel HCV therapy . A critical role of Cyclophilins , mostly P62937 ( CyPA ) , in the replication of HCV is supported by a growing body of in vitro and in vivo evidence . CyPA probably interacts directly with nonstructural protein 5A to exert its effect , through its peptidyl-prolyl isomerase activity , on maintaining the proper structure and function of the HCV replicase . The major proline substrates are located in domain II of NS5A , centered around a " DY " dipeptide motif that regulates CyPA dependence and DB00091 resistance . Importantly , DB00091 A derivatives that lack immunosuppressive function efficiently block the CyPA-NS5A interaction and inhibit HCV in cell culture , an animal model , and human trials . Given the high genetic barrier to development of resistance and the distinctness of their mechanism from that of either the current standard of care or any specifically targeted antiviral therapy for HCV ( P35610 -C ) , CyP inhibitors hold promise as a novel class of anti-HCV therapy . Behavioural and hypothalamic molecular effects of the anti-cancer agent cisplatin in the rat : A model of chemotherapy-related malaise ? Many cancer patients receiving chemotherapy experience fatigue , disturbed circadian rhythms , anorexia and a variety of dyspeptic symptoms including nausea . There is no animal model for this ' chemotherapy-related malaise ' so we investigated the behavioural and molecular effects of a potent chemotherapeutic agent , cisplatin ( CP , 6 mg/kg , i.p. ) in rats . Dark-phase horizontal locomotor activity declined post-CP reaching a nadir on day 3 ( P < 0.001 ) , before recovering after 7 days . CP 's effect was most marked in the late part ( 05.00-07.00 ) of the dark-phase . Food intake reached a nadir ( P > 0.001 ) at 2 days , coincident with an increase in gastric contents ( cisplatin 9.04+/-0.8 vs. saline 2.32+/-0.3 g ; P < 0.001 ) . No changes occurred in hypothalamic mRNA expression for O00253 , P01303 , O43612 , P06850 , IL-1 , P05231 , TNFalpha , P45844 , P31645 , P62937 and P00492 mRNA but tryptophan hydroxylase ( P17752 ) mRNA was decreased ( 47 % , P < 0.05 ) at day 21 post-CP . This shows that despite marked behavioural effects of cisplatin , only a discrete change ( P17752 ) was found in hypothalamic mRNA expression and that occurred when the animals ' behaviour had recovered . Findings are discussed in relation to the neuropharmacology of chemotherapy-induced malaise . Determination of microcystins and nodularin ( cyanobacterial toxins ) in water by LC-MS/MS . Monitoring of Lake Marathonas , a water reservoir of Athens , Greece . A method for the determination of the hepatotoxic cyanotoxins microcystins ( MCs , i.e. MC-LR , MC-RR , MC-YR , MC-LA ) and nodularin ( NOD ) in water was developed using liquid chromatography with electrospray ionization triple quadrupole mass spectrometry ( LC- P19957 -MS/MS ) after solid phase extraction ( SPE ) . New patterns of fragmentation of MC-LA were observed under the experimental conditions used . The method was fully validated to meet accreditation criteria . Mean recoveries at three concentration levels ( 0.006 , 0.1 and 1 μg L(-1) ) ranged between 70 and 114 % with % RSD values generally below 20 % . Detection limits were 2 ng L(-1) for all hepatotoxins . The method was applied to study the occurrence of MCs and NOD in Lake Marathonas , a water reservoir of Athens , over a period from July 2007 to December 2010 . The protein phosphatase inhibition assay ( P62937 ) was additionally used for fast screening of samples . MC-YR , MC-LR and MC-RR were detected and found to vary seasonally with consistent peaks during early autumn , having maximum concentrations of 717 , 451 and 174 ng L(-1) , respectively . The results of this study constitute the first report on the presence , concentration levels and seasonal variations of MCs in Lake Marathonas . None of the target cyanotoxins were detected in treated drinking water samples during the period of the study . Insights into the roles of cyclophilin A during influenza virus infection . P62937 ( CypA ) is the main member of the immunophilin superfamily that has peptidyl-prolyl cis-trans isomerase activity . CypA participates in protein folding , cell signaling , inflammation and tumorigenesis . Further , CypA plays critical roles in the replication of several viruses . Upon influenza virus infection , CypA inhibits viral replication by interacting with the M1 protein . In addition , CypA is incorporated into the influenza virus virions . Finally , DB00091 ( DB00091 ) , the main inhibitor of CypA , inhibits influenza virus replication through CypA-dependent and -independent pathways . This review briefly summarizes recent advances in understanding the roles of CypA during influenza virus infection . Immunophilins in nervous system degeneration and regeneration . Immunophilins are receptors for immunosuppressive drugs like cyclosporin A , FK506 , rapamycin and their non- immunosuppressive analogs , which are collectively referred to as " immunophilin ligands " ( Q53GA4 ) . DB00091 binds to a class of IP called cyclophilins , whereas the receptors for FK506 and rapamycin belong to the family of FK506- binding proteins ( FKBP ) . The latter are designated according to their molecular weight : P62942 , 25 , 52 etc . FKBP levels in the rat brain are up to 50 times higher than in the immune system . P62942 is associated with IP3 and ryanodine receptors present on the endoplasmic reticulum and plays a role in stabilizing calcium release . It has also been proposed to be a modulator of the TGFbeta receptor activity . Crush injury of facial or sciatic nerves in rat leads to markedly increased P62942 levels in the respective nerve nuclei and this increase is related to nerve regeneration . P62937 protects cells from death following expression of mutant Cu/ Zn superoxide dismutase , which is associated with familial amyotrophic lateral sclerosis . Our recent studies show that P62942 and Q02790 are expressed in the human nervous system , especially in the substantia nigra- deep gray matter axis . In neurodegenerative diseases , P62942 levels increase in neurons situated in areas of pathology . This IP colocalizes with synaptophysin and alpha- synuclein , suggesting that it may become a novel marker of pathology . Immunophilins participate in axonal transport , synaptic vesicle assembly and may play a role in neuroprotection against abnormal protein aggregation , suggesting a potential avenue of therapeutic interventions . P62937 is required for P61073 -mediated nuclear export of heterogeneous nuclear ribonucleoprotein A2 , activation and nuclear translocation of P27361 /2 , and chemotactic cell migration . The chemokine receptor P61073 -mediated signaling cascades play an important role in cell proliferation and migration , but the underlying mechanisms by which the receptor signaling is regulated remain incompletely understood . Here , we demonstrate that P61073 was co-immunoprecipitated with cyclophilin A ( CyPA ) from the lysate of HEK293 cells stably expressing P61073 . Although both the glutathione S-transferase- P61073 N- and C-terminal fusion proteins were associated with the purified CyPA , truncation of the C-terminal domain of P61073 robustly inhibited the receptor co-immunoprecipitation with CyPA in intact cells , thereby suggesting a critical role of the receptor C terminus in this interaction . Ligand stimulation of P61073 induced CyPA phosphorylation and nuclear translocation , both of which were inhibited by truncation of the C-terminal domain of P61073 . CyPA was associated with transportin 1 , and knockdown of transportin 1 by RNA interference ( RNAi ) blocked P48061 -induced nuclear translocation of CyPA , thereby suggesting a transportin 1-mediated nuclear import of CyPA . CyPA formed a complex with heterogeneous nuclear ribonucleoprotein ( hnRNP ) A2 , which underwent nuclear export in response to activation of P61073 . Interestingly , the P61073 -mediated nuclear export of hnRNP A2 was blocked by RNAi of CyPA . Moreover , P61073 -evoked activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) was attenuated by CyPA RNAi , by overexpression of a PPIase-deficient mutant of CyPA ( CyPA-R55A ) , and by pretreatment of the immunosuppressive drugs , cyclosporine A and sanglifehrin A . Finally , P48061 -induced chemotaxis of HEK293 cells stably expressing P61073 or Jurkat T cells was inhibited by CyPA RNAi or DB00091 treatment . NFκB inhibitors induce cell death in glioblastomas . Identification of novel target pathways in glioblastoma ( GBM ) remains critical due to poor prognosis , inefficient therapies and recurrence associated with these tumors . In this work , we evaluated the role of nuclear-factor-kappa-B ( NFκB ) in the growth of GBM cells , and the potential of NFκB inhibitors as antiglioma agents . NFκB pathway was found overstimulated in GBM cell lines and in tumor specimens compared to normal astrocytes and healthy brain tissues , respectively . Treatment of a panel of established GBM cell lines ( U138MG , U87 , U373 and P13671 ) with pharmacological NFκB inhibitors ( BAY117082 , parthenolide , MG132 , curcumin and arsenic trioxide ) and NFκB-p65 siRNA markedly decreased the viability of GBMs as compared to inhibitors of other signaling pathways such as MAPKs ( P29323 , JNK and p38 ) , PKC , P00533 and PI3K/Akt . In addition , NFκB inhibitors presented a low toxicity to normal astrocytes , indicating selectivity to cancerous cells . In GBMs , mitochondrial dysfunction ( membrane depolarization , bcl-xL downregulation and cytochrome c release ) and arrest in the G2/M phase were observed at the early steps of NFκB inhibitors treatment . These events preceded sub- P55008 detection , apoptotic body formation and caspase-3 activation . Also , NFκB was found overstimulated in cisplatin-resistant P13671 cells , and treatment of GBMs with NFκB inhibitors overcame cisplatin resistance besides potentiating the effects of the chemotherapeutics , cisplatin and doxorubicin . These findings support NFκB as a potential target to cell death induction in GBMs , and that the NFκB inhibitors may be considered for in vivo testing on animal models and possibly on GBM therapy . GNAS mutation as an alternative mechanism of activation of the Wnt/β-catenin signaling pathway in gastric adenocarcinoma of the fundic gland type . Gastric adenocarcinoma of the fundic gland type ( GAFG ) is a rare variant of gastric tumor . We have recently reported the frequent accumulation of β-catenin in GAFGs and showed that approximately half of the cases studied harbored at least 1 mutation in P35222 /AXINs/ P25054 , leading to the constitutive activation of the Wnt/β-catenin pathway . However , the mechanisms of Wnt signaling activation in the remaining cases are unknown . Accumulating evidence showed that the activating mutation in GNAS promotes tumorigenesis via the activation of the Wnt/β-catenin pathway or the P27361 /2 MAPK pathway . Therefore , we analyzed the mutations in GNAS ( exons 8 and 9 ) and in P01116 ( exon 2 ) in 26 GAFGs . Immunohistochemistry revealed nuclear β-catenin expression in 22 of 26 GAFGs , and 10 ( 38.5 % ) of 26 cases harbored at least 1 mutation in P35222 /AXINs/ P25054 . Activating mutations in GNAS were found in 5 ( 19.2 % ) of 26 GAFGs , all of which harbored R201C mutations . Activating mutations in P01116 were found in 2 ( 7.7 % ) of 26 GAFGs , and both of these also contained GNAS activating mutations . Four of 5 cases with GNAS mutation showed nuclear β-catenin expression , and presence of GNAS mutation was associated with β-catenin nuclear expression ( P = .01 ) . Furthermore , 3 of these 4 cases did not harbor mutations in P35222 , P25054 , or AXINs , suggesting that mutations in the Wnt component genes and those in GNAS occur almost exclusively . These results suggest that GNAS mutation might occur in a small subset of GAFG as an alternative mechanism of activating the Wnt/β-catenin signaling pathway . DB00091 and sanglifehrin A enhance chemotherapeutic effect of cisplatin in P13671 glioma cells . Glioma is the most common type of brain tumors in adults , and treatment of high-grade gliomas is still palliative . Studies to date have revealed only modest effect in attenuating growth of these tumors with single agent therapy , but combination treatment appears to be more effective . P62937 ( CypA ) , a target of immunosuppressive drugs cyclosporin A ( DB00091 ) and sanglifehrin A ( SFA ) , is an intracellular protein that has peptidyl-prolyl cis-trans isomerase ( PPIase ) enzymatic activity . Previously , we showed that overexpressed CypA induced chemoresistance in cancer cells . Here we provide evidence that combination of cisplatin with either DB00091 or SFA synergistically enhances apoptotic cell death in P13671 glioma cells , compared with single agent treatment . Enhanced apoptotic cell death is a result of an increase in ROS generation and a decrease in intracellular glutathione levels . Consistently , CypA knockdown by siRNA also enhances cisplatin-induced apoptosis . Immunohistochemical analysis showed increased expression of CypA in human glioblastoma multiforme , but not in normal human astrocytes . CypA was also shown to be up-regulated in P13671 glioma cells during hypoxia . In conclusion , DB00091 or SFA in combination with cisplatin synergistically enhances cisplatin-induced apoptosis in P13671 glioma cells via inhibition of PPIase activity of CypA , indicating that development of new drugs that selectively inhibit the CypA PPIase activity without immune suppression may facilitate alleviation of chemoresistance in treatment of high-grade glioma . Oxidative stress induces extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase in cystic fibrosis lung epithelial cells : Potential mechanism for excessive P10145 expression . Cystic fibrosis ( CF ) is a lethal disease caused by defective function of the cftr gene product , the CF transmembrane conductance regulator ( P13569 ) that leads to oxidative damage and excessive inflammatory response in lungs of CF patients . We here report the effects of oxidative stress ( hyperoxia , 95 % O(2) ) on the expression of pro-inflammatory interleukin ( IL ) -8 and P25024 /2 receptors in two human CF lung epithelial cell lines ( IB3-1 , with the heterozygous F508del/W1282X mutation and CFBE41o- with the homozygous F508del/F508del mutation ) and two control non-CF lung epithelial cell lines ( S9 cell line derived from IB3-1 after correction with wtCFTR and the normal bronchial cell line 16HBE14o- ) . Under oxidative stress , the expression of P10145 and P25024 /2 receptors was increased in CF , corrected and normal lung cell lines . The effects of oxidative stress were also investigated by measuring the transcription nuclear factor kappaB ( NF-kappaB ) and activator protein-1 ( AP-1 ) activities . Under oxidative stress , no increase of NF-kappaB activation was observed in CF lung cells in contrast to that observed in normal and corrected CF lung cells . The signalling of mitogen-activated protein ( Q96HU1 ) kinases was further studied . We demonstrated that extracellular signal-regulated kinase ( P27361 /2 ) and AP-1 activity was markedly enhanced in CF but not non-CF lung cells under oxidative stress . Consistently , inhibition of P27361 /2 in oxidative stress-exposed CF lung cells strongly decreased both the P10145 production and P25024 /2 expression . Therefore , targeting of P27361 /2 Q96HU1 kinase may be critical to reduce oxidative stress-mediated inflammation in lungs of CF patients . [ Low doses of sulphonyluria as a successful replacement for insulin therapy in a patient with neonatal diabetes due to a mutation of Q14654 gene encoding Kir6.2 ] . Neonatal diabetes mellitus is a rare metabolic disorder with an estimated incidence of 1:300.000 to 400.000 newborns , and less than 50 % of the neonates have permanent neonatal diabetes mellitus ( PNDM ) . Recently , activating mutation in the Q14654 gene encoding Kir6.2 subunit of the adenosin triphosphate-sensitive potassium ( K( DB00171 ) ) channel has been described as the most frequent cause of PNDM . Under physiological circumstances K( DB00171 ) channel closure plays a central role in glucose-stimulated insulin secretion from pancreatic beta cells . Sulphonylurea drugs stimulate insulin secretion by binding to and closing K( DB00171 ) channels and thus bypassing beta cell metabolism stimulate the same chain of reactions as glucose . We describe a boy diagnosed with PNDM at the age of 3 months when insulin therapy was started , and at the age of 4.5 years Q14654 gene was sequenced and found that the boy carried a de novo activating R201H mutation . P01308 therapy was successfully switched to low doses of oral glibenclamide . Accordingly , it is important to emphasize that every person diagnosed with diabetes before six months of life , however old they actually are , should be tested for K( DB00171 ) mutations which is offered via the website www.diabetesgenes.org . Effect of cyclooxygenase inhibitors on the P06850 -induced pituitary-adrenocortical activity during crowding stress . The aim of the present study was to determine the effect of social stress and significance of prostaglandins ( PG ) generated by constitutive and inducible cyclooxygenase ( P23219 and P35354 ) in the stimulation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis by corticotropin releasing hormone ( P06850 ) under basal and social crowding stress conditions . The stressed rats were crowded in groups of 24 to a cage for 3 or 7 days , whereas the control animals were haused in groups of 7 to a cage of the same size . The activity of Q9Y251 axis was determined by measuring plasma DB01285 and serum corticosterone levels 1 h after i.p . P06850 administration . Inhibitors of P23219 , piroxicam ( 0.2 , 2.0 , and 5.0 mg/kg ) , and P35354 , compound NS-398 ( 0.2 and 2.0 mg/kg ) , were administered i.p . 15 min prior to P06850 ( 0.1 microg/kg i.p. ) to control or crowded rats . The obtained results indicate that social stress for 3 and 7 days markedly intensifies the stimulatory action of P06850 on DB01285 secretion . Neither piroxicam nor NS-398 induce any significant effect on the P06850 -elicited DB01285 and corticosterone secretion in non-stressed or crowded rats . Therefore , PG generated by P23219 or P35354 do not participate to a significant extent in the stimulation of Q9Y251 axis by P06850 under either basal conditions or during crowding stress . These results also indicate that the stimulatory action of P06850 on DB01285 secretion is not only completely resistant to desensitization but is sensitized during social crowding stress . The results contrast with a significant involvement of PG in the vasopressin-induced stimulation of Q9Y251 response during crowding stress . Distinct pathways involving the FK506-binding proteins 12 and 12.6 underlie P60568 -versus P40933 -mediated proliferation of T cells . The molecular basis for the different roles of P60568 and P40933 in lymphocyte function has been poorly defined . Searching for differences that underlie the distinct T cell responses to the two cytokines , we observed a marked susceptibility of the P40933 -induced but not of the P60568 -induced proliferation to rapamycin despite a decrease of p70S6 kinase ( p70S6K ) activation by the drug in response to both cytokines . Activated splenic T lymphocytes deficient in the FK506-binding protein ( FKBP ) 12 , a target of rapamycin activity , had reduced proliferation in response to P40933 but not to P60568 . This decreased proliferation was accompanied by reduced activation of p70S6K and of the extracellular signal-regulated kinases ( P29323 ) after P40933 treatment . In contrast to P62942 -/- cells , splenic P68106 -/- T cells exhibited a decreased proliferative response to P60568 in the presence of rapamycin without affecting p70S6K or P29323 activation . Thus , P40933 induces T cell proliferation mainly via P62942 -mediated p70S6K activation . In contrast , P60568 signaling involves multiple pathways that include at least one additional pathway that depends on P68106 . The non-immunosuppressive cyclosporin A analogue SDZ NIM 811 inhibits cyclophilin A incorporation into virions and virus replication in human immunodeficiency virus type 1-infected primary and growth-arrested T cells . SDZ NIM 811 is a cyclosporin A ( DB00091 ) analogue that is completely devoid of immunosuppressive capacity but exhibits potent and selective anti-human immunodeficiency virus type 1 ( HIV-1 ) activity . Binding to cyclophilin A , the intracellular receptor for cyclosporins , is a prerequisite for HIV-1 inhibition by cyclosporins . P62937 was demonstrated to bind to HIV-1 p24gag and this cyclophilin-Gag interaction leads to the incorporation of cyclophilin A into HIV-1 virions . SDZ NIM 811 inhibits this protein interaction , and this is likely to be the molecular basis for its antiviral activity . Here , we show that in activated primary T cells SDZ NIM 811 interferes with two stages of the virus replication cycle : ( i ) translocation of pre-integration complexes into the nucleus and ( ii ) production of infectious virus particles . SDZ NIM 811 not only inhibits translocation of HIV-1 pre-integration complexes in primary T cells , but also in a growth-arrested T cell line . In vivo , most T lymphocytes are quiescent , but serve nevertheless as a major and inducible HIV-1 reservoir in infected individuals . Significant amounts of cyclophilin A were found to be associated with virus particles propagated in primary T cells . SDZ NIM 811 caused a strong reduction in the amount of incorporated cyclophilin A , thereby reducing infectivity . Thus , cyclophilin A seems to be necessary for HIV-1 replication in primary T cells . 1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea : a novel cyclophilin A allosteric activator . P62937 ( CypA ) plays an important role in many physiology processes and its overexpression has been involved in many diseases including immune disease , viral infection , neuro-degenerative disease , and cancer . However , the actual role of CypA in the diseases is still far from clear , and a complete understanding of CypA is necessary in order to direct more specific and effective therapeutic strategies . Based on the screening of our in-house library through the isomer-specific proteolysis method , we find a CypA activator ( 1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea ) , compound 1a , which can increase CypA 's PPIase activity and give allosteric behavior . The binding affinity of compound 1a to CypA has been confirmed by Fortebio 's Octet Q13123 system and the increased phosphorylation of P29323 in H446 cells is observed by treatment with both compound 1a and DB00091 . In order to further evaluate the binding mode between the activator and CypA , the allosteric binding site and allosteric mechanism of CypA are investigated by molecular dynamics ( MD ) simulations in combination with mutagenesis experiments . The results show that the allosteric binding site of CypA is 7Å away from its catalytic site and is composed of Cys52 , His70 , His54 , Lys151 , Thr152 and Lys155 . Compound 1a binds to the allosteric site of CypA , stabilizing the active conformation of catalytic residues , and finally promotes the catalytic efficiency of CypA . We believe our finding of the CypA allosteric activator will be used as an effective chemical tool for further studies of CypA mechanisms in diseases . P62937 is upregulated in small cell lung cancer and activates P27361 /2 signal . P62937 ( CypA ) , a peptidyl-prolyl cis-trans isomerase ( PPIase ) , was originally identified as the intracellular receptor for cyclosporin A ( DB00091 ) . Recently , correlations of CypA with tumor pathogenesis have been studied . Here , we studied the expression of CypA and its receptor CD147 in several kinds of lung cancer cells as well as a normal lung cell and found that in H446 cell , a kind of small cell lung cancer cell , the expression are the highest . The exogeneous CypA protein can substantially stimulate H446 cell growth in dependence on its PPIase activity . We also showed that CypA protein can stimulate P27361 /2 signal in dose and time dependent manners and almost has no effect to p38 and JNK signals . Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for small cell lung cancer . Q96RP3 is expressed in human pregnant myometrial cells and regulates myosin light chain phosphorylation : potential role of the type-2 corticotropin-releasing hormone receptor in the control of myometrial contractility . The family of P06850 -related peptides are suggested to play important roles in the control of myometrial contractility during pregnancy and labor . In this study we investigated the expression of urocortin II ( P55089 II ) in human myometrium and its ability to phosphorylate intracellular components that can be involved in modulating myometrial contractility . Using RT-PCR and fluorescent in situ hybridization , we demonstrated that P55089 II and type-2 P06850 receptor ( Q13324 ) mRNAs were expressed in human nonpregnant and pregnant myometrium . Immunofluorescent studies confirmed protein expression of P55089 II in human pregnant myometrial cells , whereas chemical cross-linking studies with radiolabeled P55089 II confirmed the presence of Q13324 sites with an apparent molecular mass of 50 kDa . Treatment of primary human myometrial cells with P55089 II to specifically activate Q13324 resulted in a dose-dependent increase of myosin light chain ( MLC(20) ) phosphorylation . Activation of protein kinase C ( PKC ) and P27361 /2 was required for the P55089 II-induced activation of MLC(20) , because treatment of myometrial cells with inhibitors of MAPK kinase 1 ( U0126 ) and PKC ( bisindolylmaleimide ) inhibited the P55089 II-induced phosphorylation of MLC(20) . Furthermore , the P55089 II effect on MLC(20) was dependent on RhoA translocation to the membrane and subsequent activation of RhoA-associated kinase , as shown by the use of the specific inhibitors exoenzyme P01024 and Y27632 . Collectively , our data suggest a distinctive role for Q13324 - specific agonists like P55089 II in the control of myometrial contractility during human pregnancy involving sequential activation of PKC , MAPK kinase 1 , P27361 /2 , RhoA , and RhoA-associated kinase , leading to the MLC(20) phosphorylation . P62937 is an inflammatory mediator that promotes atherosclerosis in apolipoprotein E-deficient mice . P62937 ( CyPA ; encoded by Ppia ) is a ubiquitously expressed protein secreted in response to inflammatory stimuli . CyPA stimulates vascular smooth muscle cell migration and proliferation , endothelial cell adhesion molecule expression , and inflammatory cell chemotaxis . Given these activities , we hypothesized that CyPA would promote atherosclerosis . P02649 -deficient ( Apoe(-/-) ) mice fed a high-cholesterol diet for 16 wk developed more severe atherosclerosis compared with Apoe(-/-)Ppia(-/-) mice . Moreover , CyPA deficiency was associated with decreased low-density lipoprotein uptake , P19320 ( vascular cell adhesion molecule 1 ) expression , apoptosis , and increased P29474 ( endothelial nitric oxide synthase ) expression . To understand the vascular role of CyPA in atherosclerosis development , bone marrow ( BM ) cell transplantation was performed . Atherosclerosis was greater in Apoe(-/-) mice compared with Apoe(-/-)Ppia(-/-) mice after reconstitution with CyPA(+/+) BM cells , indicating that vascular-derived CyPA plays a crucial role in the progression of atherosclerosis . These data define a role for CyPA in atherosclerosis and suggest CyPA as a target for cardiovascular therapies . Marine invertebrates cross phyla comparisons reveal highly conserved immune machinery . Naturally occurring histocompatibility responses , following tissue-to-tissue allogeneic contacts , are common among numerous colonial marine invertebrate taxa , including sponges , cnidarians , bryozoans and ascidians . These responses , often culminating in either tissue fusions or rejections , activate a wide array of innate immune components . By comparing two allorejection EST libraries , developed from alloincompatible challenged colonies of the stony coral Stylophora pistillata and the ascidian Botryllus schlosseri , we revealed a common basis for innate immunity in these two evolutionary distant species . Two prominent genes within this common basis were the immunophilins , P62937 ( CypA ) and FK506-binding protein ( FKBP ) . In situ hybridizations revealed that mRNA expression of the coral and ascidian immunophilins was restricted to specific allorecognition effector cell populations ( nematoblasts and nematocytes in the coral and morula cells in the ascidian ) . The expressions were limited to only some of the effector cells within a population , disclosing disparities in numbers and location between naïve colonies and their immune challenged counterparts . Administration of the immunosuppression drug DB00091 -A during ascidian 's allogeneic assays inhibited both fusion and rejection reactions , probably through the inhibition of ascidian 's immunocytes ( morula cells ) movement and activation . Our results , together with previous published data , depict an immunophilins-based immune mechanism , which is similarly activated in allogeneic responses of distantly related animals from sponges to humans . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Design and synthesis of conformationally constrained cyclophilin inhibitors showing a cyclosporin-A phenotype in C. elegans . P62937 ( CypA ) is a member of the immunophilin family of proteins and receptor for the immunosuppressant drug cyclosporin A ( DB00091 ) . Here we describe the design and synthesis of a new class of small-molecule inhibitors for CypA that are based upon a dimedone template . Electrospray mass spectrometry is utilised as an initial screen to quantify the protein affinity of the ligands . Active inhibitors and fluorescently labelled derivatives are then used as chemical probes for investigating the biological role of cyclophilins in the nematode Caenorhabditis elegans .

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interacts_with DB00831?

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First report of warfarin dose requirements in patients possessing the P11712 *12 allele . BACKGROUND : DB00682 is the most frequently prescribed anticoagulant in North America and Europe . It is administered as a racemate , but S-warfarin is principally responsible for its anticoagulant activity . Cytochrome P450 ( CYP ) 2C9 is the enzyme primarily responsible for the metabolism of S-warfarin . Numerous variant alleles of P11712 have been identified . The P11712 *12 ( rs9332239 ) allele harbors a P489S substitution in P11712 which has been shown to result in a 40 % decline in catalytic activity in vitro . CASES : Four Caucasian patients with a low mean weekly warfarin dose ( MWWD ) were genotyped for P11712 , Q9BQB6 and P02649 variant alleles . None of the four patients carried the common P11712 variant alleles ( *2 , *3 , *5 , *6 , *7 , *8 , *9 , *11 , *13 ) despite a relatively low MWWD ( 23.4±7.94 mg ) compared to 208 patients carrying the CYP29C9*1 genotype ( 32.2±12.65 mg ) . Given that P11712 *12 confers decreased in vitro activity to the enzyme , we investigated whether these patients carried this allele . All four patients were P11712 *12 CT heterozygotes . Individual comparisons with patients possessing the same Q9BQB6 and P02649 genotypes also demonstrated lower dose requirements in the patients that possessed P11712 *12 allele . CONCLUSIONS : There are no reports of the clinical impact of rs9332239 on P11712 substrates . This is the first report of patients with the rare P11712 *12 genotype and lower warfarin dose requirements . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . [ P62158 -dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium ] . Highly purified plasma membrane ( PM ) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein . Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3 % of the original level . The activity of Ca , Mg-ATPase is thereby decreased by 40 % . Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour . The maximal activation of Ca , Mg-ATPase is observed with 10(-7) M calmodulin . P62158 increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity . DB00831 ( 20 microM ) diminishes the activating effect of exogenous calmodulin on Ca , Mg-ATPase . P62158 stimulates Ca , Mg-ATPase at low concentrations of Ca2+ ( 10(-8)-10(-6) M ) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax -- from 0,8 to 1.42 mumol Pl/mg protein/hour . It is supposed that the activating effect of calmodulin on Ca , Mg-ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme . Adhesion molecules , mycophenolate mofetil and systemic lupus erythematosus . DB00688 ( DB00688 ) has been reproducibly shown to inhibit lymphocyte adhesion and penetration of endothelial cell surfaces . The mechanism is not yet elucidated . In vitro studies on the effects of DB00688 on cell adhesion molecules ( P62158 ) using human umbilical vein endothelial cells ( HUVEC ) have shown conflicting results . Different studies have independently shown that DB00688 increased , decreased or had no effect on intercellular adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule ( P19320 ) . Several studies suggest DB00688 may reduce the endothelial expression of P16581 . Recent studies have been unable to replicate initial work , which suggested that DB00688 impaired glycosylation of lymphocyte P62158 . The same studies concluded that DB00688 had no effect on the surface expression of lymphocyte P62158 , but altered the binding ability of these molecules . P05362 /LFA-1 ( lymphocyte function-associated antigen-1 ) , P19320 /VLA-4 ( very late antigen-4 ) and P16109 / Q14242 ( P16109 glycoprotein ligand-1 ) ligand pairs are most likely to be involved . Few in vivo and no conclusive human studies have been carried out . The literature relevant to cell adhesion molecules in systemic lupus erythematosus ( SLE ) is reviewed in detail . Oral clarithromycin enhances airway immunoglobulin A ( IgA ) immunity through induction of IgA class switching recombination and Q9Y275 of the tumor necrosis factor family molecule on mucosal dendritic cells in mice infected with influenza A virus . We previously reported that the macrolide antibiotic clarithromycin ( P62158 ) enhanced the mucosal immune response in pediatric influenza , particularly in children treated with the antiviral neuraminidase inhibitor oseltamivir ( OSV ) with low production of mucosal antiviral secretory IgA ( S-IgA ) . The aims of the present study were to confirm the effects of P62158 on S-IgA immune responses , by using influenza A virus ( IAV ) H1N1-infected mice treated with or without OSV , and to determine the molecular mechanisms responsible for the induction of mucosal IgA class switching recombination in IAV-infected P62158 -treated mice . The anti-IAV S-IgA responses and expression levels of IgA class switching recombination-associated molecules were examined in bronchus-lymphoid tissues and spleens of infected mice . We also assessed neutralization activities of S-IgA against IAV . Data show that P62158 enhanced anti-IAV S-IgA induction in the airway of infected mice and restored the attenuated antiviral S-IgA levels in OSV-treated mice to the levels in the vehicle-treated mice . The expression levels of Q9Y275 of the tumor necrosis factor family ( Q9Y275 ) molecule on mucosal dendritic cells as well as those of activation-induced cytidine deaminase and Iμ-Cα transcripts on B cells were enhanced by P62158 , compared with the levels without P62158 treatment , but P62158 had no effect on the expression of the Q96RJ3 on B cells . Enhancement by P62158 of neutralization activities of airway S-IgA against IAV in vitro and reinfected mice was observed . This study identifies that P62158 enhances S-IgA production and neutralizing activities through the induction of IgA class switching recombination and upregulation of Q9Y275 molecules in mucosal dendritic cells in IAV-infected mice . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Synergistic inhibition of angiogenesis by artesunate and captopril in vitro and in vivo . Inhibition of angiogenesis represents one major strategy of cancer chemotherapy . In the present investigation , we investigated the synergism of artesunate and captopril to inhibit angiogenesis . DB09274 is an antimalarial derivative of artemisinin from the Chinese medicinal plant , Artemisia annua L. , which also reveals profound anticancer activity in vitro and in vivo . DB01197 is an angiotensin I-converting ( P12821 ) inhibitor , which is well established in Western academic medicine . Both compounds inhibited migration of human umbilical vein endothelial cells ( HUVECs ) in vitro . The combination of both drugs resulted in synergistically inhibited migration . Whereas artesunate inhibited HUVEC growth in the XTT assay , captopril did not , indicating independent modes of action . We established a chorioallantoic membrane ( P62158 ) assay of quail embryos ( Coturnix coturnix L. ) and a computer-based evaluation routine for quantitative studies on vascularization processes in vivo . DB09274 and captopril inhibited blood vessel formation and growth . For the first time , we demonstrated that both drugs revealed synergistic effects when combined . These results may also have clinical impact , since cardiovascular diseases and cancer frequently occur together in older cancer patients . Therefore , comorbid patients may take advantage , if they take captopril to treat cardiovascular symptoms and artesunate to treat cancer . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Skinned coronary smooth muscle : calmodulin , calcium antagonists , and DB02527 influence contractility . The effects of Ca2+ , calmodulin , DB02527 , the catalytic subunit of DB02527 -dependent protein kinase ( CSU ) and some Ca2+ antagonists were studied in chemically ( Triton X-100 ) skinned coronary smooth muscle . P62158 increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration . DB00831 , a calmodulin antagonist , inhibited Ca2+-calmodulin-induced contraction . Both DB02527 and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration . DB08980 , a Ca2+-antagonist , at 2 x 10(-4) M produced a significant inhibitory effect , which was reduced by increasing the Ca2+ concentration . From other Ca2+ antagonists tested , W-7 , but not D600 and verapamil , produced some inhibitory effect . The data indicate that the response of skinned coronary smooth muscle to Ca2+ , calmodulin and DB02527 are similar to those obtained with other skinned smooth muscles . Furthermore , skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle . Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . Anti-angiogenic action of 5,5-diphenyl-2-thiohydantoin-N10 ( DPTH-N10 ) . Previously , we demonstrated that 5,5-diphenyl-2-thiohydantoin ( DPTH ) exerts an anti-proliferation effect on subcultured human umbilical vein endothelial cells ( HUVEC ) . In the present study , we show that 2(naphthalen-2-ylmethylsulfanyl)-5,5-diphenyl-1,5-dihydro-imidazol-4-one ( DPTH-N10 ) , a derivative compound of DPTH , exerts a 5 times stronger inhibition of [3H]thymidine incorporation into HUVEC as compared with DPTH and at very low concentrations ( 0-20 microM ) inhibited DNA synthesis and decreased cell number in cultured HUVEC in a concentration- and time-dependent manner , but not in human fibroblasts . [3H]thymidine incorporation analysis demonstrated that treatment of HUVEC with DPTH-N10 arrested the cell at the G0/ P55008 phase of the cell cycle . Western blot analysis revealed that the protein level of P38936 in HUVEC increased after DPTH-N10 treatment . In contrast , the protein levels of p27 , p53 , cyclins A , D1 , D3 and E , cyclin-dependent kinase (CDK)2 , and P11802 in HUVEC were not changed significantly after DPTH-N10 treatment . Immunoprecipitation showed that the formation of the P24941 - P38936 complex , but not the P24941 -p27 , P11802 - P38936 , and P11802 -p27 complex , was increased in the DPTH-N10-treated HUVEC . Kinase assay further demonstrated that P24941 , but not P11802 , kinase activity was decreased in the DPTH-N10-treated HUVEC . Pretreatment of HUVEC with a P38936 , but not p27 , antisense oligonucleotide reversed the DPTH-N10-induced inhibition of [3H]thymidine incorporation into HUVEC . Taken together , these data suggest that DPTH-N10 inhibits HUVEC proliferation by increasing the level of P38936 protein , which in turn inhibits P24941 kinase activity , and finally interrupts the cell cycle . Capillary-like tube formation , aortic ring culture , and chick embryo chorioallantoic membrane ( P62158 ) assays further demonstrated the anti-angiogenic effect of DPTH-N10 . Targeting the integrated networks of aggresome formation , proteasome , and autophagy potentiates ER stress‑mediated cell death in multiple myeloma cells . The inhibitory effects of macrolide antibiotics including clarithromycin ( P62158 ) on autophagy flux have been reported . Although a macrolide antibiotic exhibits no cytotoxicity , its combination with bortezomib ( BZ ) , a proteasome inhibitor , for the simultaneous blocking of the ubiquitin (Ub)‑proteasome and autophagy‑lysosome pathways leads to enhanced multiple myeloma ( MM ) cell apoptosis induction via stress overloading of the endoplasmic reticulum ( ER ) . As misfolded protein cargo is recruited by histone deacetylase 6 ( Q9UBN7 ) to dynein motors for aggresome transport , serving to sequester misfolded proteins , we further investigated the cellular effects of targeting proteolytic pathways and aggresome formation concomitantly in MM cells . Pronounced apoptosis was induced by the combination of vorinostat [ suberoylanilide hydroxamic acid ( DB02546 ) ; potently inhibits Q9UBN7 ] with P62158 and BZ compared with each reagent or a 2‑reagent combination . P62158 /BZ treatment induced vimentin positive‑aggresome formation along with the accumulation of autolysosomes in the perinuclear region , whereas they were inhibited in the presence of DB02546 . The DB02546 / P62158 /BZ combination treatment maximally upregulated genes related to ER stress including C/EBP homologous protein ( P35638 ) . Similarly to MM cell lines , enhanced cytotoxicity with P35638 upregulation following DB02546 / P62158 /BZ treatment was shown by a wild‑type murine embryonic fibroblast ( MEF ) cell line ; however , a CHOP‑deficient MEF cell line almost completely canceled this pronounced cytotoxicity . Knockdown of Q9UBN7 with siRNA exhibited further enhanced P62158 /BZ‑induced cytotoxicity and P35638 induction along with the cancellation of aggresome formation . Targeting the integrated networks of aggresome , proteasome , and autophagy is suggested to induce efficient ER stress‑mediated apoptosis in MM cells . Association between iris constitution and apolipoprotein e gene polymorphism in hypertensives . OBJECTIVE : Iridology is a complementary and alternative medicine ( P62158 ) that involves the diagnosis of medical conditions by noting irregularities of the pigmentation in the iris . Iris constitution has a strong familial aggregation and heredity is implicated . P02649 ( apoE ) gene polymorphism is one of the most well-studied genetic markers for vascular diseases , including hypertension . In this study , we investigated the relationship between iris constitution and apoE polymorphism in hypertensives . DESIGN AND SUBJECTS : We classified 87 hypertensives and 79 controls according to iris constitution and determined the apoE genotype of each individual . RESULTS : A significantly higher percentage of individuals with neurogenic constitutions was found in the hypertensive group when compared with the control group ( chi(2) = 40.244 , p < 0.001 ) . In addition , a neurogenic constitution increased the relative risk for hypertension for subjects with an apo epsilon2 or an epsilon4 allele ( chi(2) = 4.086 , p = 0.049 , odds ratio = 2.633 , confidence interval = 1.004-6.905 ) . CONCLUSIONS : Our results imply that a neurogenic iris constitution enhances the relative risk for hypertension in subjects with the apo epsilon2 or epsilon4 allele . Furthermore , we attempted to evaluate the efficacy of iris constitutional medicine and to find an association with hypertension .

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DB00624 and androgen receptor in human nephrolithiasis . PURPOSE : We investigated the relationship of kidney calculi with plasma free and total testosterone , and androgen receptor up-regulation in the kidneys of men with nephrolithiasis . MATERIALS AND METHODS : Male patients with kidney stone and healthy men were included in the study . Blood was collected in a tube containing 2 % heparin in the morning . Total and free serum testosterone was measured by enzyme linked immunosorbent assay . All patients underwent percutaneous nephrostolithotomy . At the end of the procedure ultrasound guided puncture biopsy was done to acquire kidney tissue . Normal kidney tissue obtained at autopsy served as the control . P10275 was detected in kidney tissue by immunohistochemistry . Stone composition was analyzed in each patient . RESULTS : The study included 37 male patients 22 to 39 years old and 31 healthy men 24 to 37 years old . All calculi were composed of calcium oxalate monohydrate or calcium oxalate dihydrate and a few also contained protein or uric acid . Mean±SD serum total and free testosterone was 13.29±4.79 ng/ml and 63.23±28.58 pg/ml in patients , and 7.30±0.82 ng/ml and 35.59±24.91 pg/ml in healthy men , respectively ( each p < 0.001 ) . Immunohistochemistry revealed androgen receptor up-regulation in the kidneys of patients with nephrolithiasis . CONCLUSIONS : Our data suggest the important role of enhanced androgen signaling in human nephrolithiasis . P10275 is essential for sexual differentiation of responses to olfactory cues in mice . During sexual differentiation males and females are exposed to different levels of testosterone , which promotes sex differences in the adult brain and in behavior . DB00624 can act after aromatization or reduction via a number of steroid hormone receptors . Here we provide new evidence that the androgen receptor ( AR ) is essential for sexual differentiation in mice . We used mice carrying the testicular feminization ( Tfm ) mutation of the AR . Adult Tfm males , wild-type male and female littermates were gonadectomized and given subcutaneous estradiol implants . In all sexually dimorphic traits , Tfm males had responses equivalent to females and different from males . In simultaneous choice tests , males spent significantly more time investigating female-soiled bedding , whereas females and Tfm males preferred to investigate male-soiled bedding . Tfm males and females did not have a partner preference in tests with awake stimulus animals , whereas males showed a preference for females over males . Exposure to male-soiled , but not clean , bedding produced a significant increase in c-Fos-immunoreactive cells in the medial preoptic area and bed nucleus of the stria terminalis in Tfm males and females , no increase was noted in males . Masculine sexual behavior ( mounting and thrusting ) was not sexually dimorphic , and all groups displayed these behaviors . Our results support data collected in humans suggesting a role for the androgen receptor in sexual differentiation of social preferences and neural responses to pheromones . Gender-specific effects of endogenous testosterone : female alpha-estrogen receptor-deficient C57Bl/6J mice develop glomerulosclerosis . Young female mice on a C57Bl/6J ( B6 ) background are considered glomerulosclerosis ( GS ) -resistant but aging B6 mice develop mild GS . Estrogen deficiency accelerates while estrogen replacement retards GS in young sclerosis-prone oligosyndactyly mutant mice on an P04000 background . To explore the effects of sex hormones on glomerular structure and function in the context of gender and genetic background , we studied mice in which the estrogen-receptor ( ER ) genes alpha- or -beta were deleted ( alpha- or betaER knockout ( KO ) ) and crossed into the B6 background . We also studied ovariectomized ( Ovx ) B6 mice given testosterone . Male and female betaERKO and male alphaERKO mice had no glomerular dysfunction at 9 months of age ; however , alphaERKO female mice displayed albuminuria and GS . Ovx prevented glomerular dysfunction in alphaERKO female mice by eliminating endogenous testosterone production while exogenous testosterone induced GS in Ovx B6 mice . P10275 ( AR ) expression and function was found in microdissected glomeruli and cultured mesangial cells . DB00624 compared to placebo increased both AR expression and TGF-beta1 mRNA levels in glomeruli isolated from female B6 mice . Estrogen deficiency had no deleterious effects on the glomeruli in B6 mice . Our study shows that genetic traits strongly influence the GS-promoting effects of estrogen deficiency while testosterone induces GS in a gender-specific manner . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Androgens and metabolic syndrome : lessons from androgen receptor knock out ( ARKO ) mice . DB00624 ( T ) is an important factor for determining body composition in males . Abdominal obesity is inversely correlated with serum T levels in men , leading to greater mortality . Pathologically hypogonadal men also have a significantly higher fat mass , which is reversed by T administration . However , the mechanism for such anti-obesity effect of androgen has not been well clarified . P10275 ( AR ) null male mice revealed late-onset obesity . Male ARKO mice were euphagic compared to the wild-type male controls , but also less dynamic and less oxygen consuming . Transcript profiling indicated that male ARKO mice had lower transcripts for the thermogenetic uncoupling protein 1 ( P25874 ) . We also found enhanced secretion of adiponectin , which is insulin-sensitizing , from adipose tissue in comparison to wild type , which might partly explain why the overall insulin sensitivity of male ARKO mice remained almost intact despite their apparent obesity . In addition , decreased lipolysis rather than increased lipid synthesis was observed , which might also account for the increased adiposity in male ARKO mice . The results revealed that AR plays important roles in male metabolism by affecting the energy balance , and is negative to both adiposity and insulin sensitivity . Neonatal estrogen exposure disrupts uterine development in the postnatal sheep . Postnatal development of the ovine uterus between birth and postnatal day ( P01160 ) 56 involves budding differentiation of the endometrial glandular epithelium from the luminal epithelium ( LE ) followed by extensive coiling and branching morphogenesis of the tubular glands . To determine the short- and long-term effects of estrogen on neonatal ovine uterine development after P01160 14 , neonatal sheep were randomly assigned at birth ( P01160 0 ) to be treated daily with estradiol-17beta benzoate ( EB ; 0 , 0.01 , 0.1 , 1 , or 10 microg/kg body weight.d ) during one of two developmental periods ( P01160 14-27 or 42-55 ) . All ewes were hemiovariohysterectomized at the end of EB treatment on either P01160 28 or 56 , and the remaining uterine horn and ovary removed on P01160 112 . Immediate responses to EB treatment included dose- and age-dependent increases in uterine wet weight , thickness of the endometrium , myometrium , and LE , but decreases in endometrial glands on P01160 28 and 56 . Transient exposure to EB decreased gland number and thickness of the endometrium and LE on P01160 112 but did not affect extrauterine reproductive tract structures . The mechanism of estrogen inhibition of uterine development did not involve effects on cell proliferation . Real-time PCR analyses found that EB exposure disrupted normal patterns of growth factor ( P05019 , P01344 , fibroblast growth factor-7 , fibroblast growth factor-10 , and hepatocyte growth factor ) and receptor mRNA expression in the uterus . Transient exposure of the neonatal ewe to estrogens during critical periods specifically alters growth factor networks that perturb normal development of the uterus , leading to permanent alterations in uterine structure and function . The androgen receptor : a mediator of diverse responses . Androgens mediate a number of diverse responses through the androgen receptor , a 110 kD ligand-activated nuclear receptor . P10275 expression , which is found in a variety of tissues , changes throughout development , aging , and malignant transformation . The androgen receptor can be activated by two ligands , testosterone and dihydrotestosterone , which bind to the androgen receptor with different affinities . This difference in binding affinity results in different levels of activation of the androgen receptor by the two ligands . The androgen receptor acts as a transcriptional modifier of a variety of genes by binding to an androgen response element . The ability to confer androgen specific actions by the androgen response element may depend on other cell-specific transcription factors and cis-acting DNA elements in close proximity to it . DB00624 and dihydrotestosterone appear to act upon an identical nuclear receptor . However , in certain instances , they mediate different physiologic responses . For example , dihydrotestosterone , but not testosterone , is capable of mediating full sexual development of the male external genitalia . In some cases , the androgen receptor may induce opposite physiologic responses in similar tissue types depending on their location . For example , in male pattern baldness , activated androgen receptors may suppress the growth of distinct hair follicle populations through initiating stromal-epithelial actions , whereas other hair follicles continue to proliferate . In other cases , altered androgen receptor activity due to its mutation or altered expression may lead to pathology such as recurrence of prostate cancer due to development of androgen independence allowing tumor cell proliferation under androgen deprivation . P10275 dependent and independent activities of testosterone on hepatic microsomal drug metabolism . Administration of testosterone for 6 days to intact female and castrate male BALB/cJ mice stimulated hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content by 50-75 % . DB00624 also stimulated hepatic microsomal NADPH-oxidase activity , but to a lesser degree . To probe the mechanism of this effect of androgens , two antiandrogens ( cyproterone acetate and flutamide ) were employed . Since cyproterone acetate was a potent stimulator of hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content , no antiandrogenic activity of this steroid could be detected . By contrast , flutamide alone had little effect on either ethylmorphine N-demethylase activity or cytochrome P-450 content . However , this drug effectively blocked the stimulatory effects of testosterone on ethylmorphine N-demethylase activity and cytochrome P-450 content but not on NADPH-oxidase activity . This effect was not species specific , since flutamide also prevented androgen stimulation of ethylmorphine metabolism in adult castrate and prepubertal male Fisher rats . The testosterone-induced increase of hepatic weight and microsomal protein content was not affected by the administration of flutamide . The observations are consistent with the hypothesis that androgens have two distinct effects on the liver . First , testosterone may act as a general , nonspecific stimulant of liver weight and microsomal protein content which is independent of the androgen receptor . Secondly , testosterone action in the liver may be expressed via an androgen-specific or androgen receptor-dependent mechanism which controls , in part , the cytochrome P-450-dependent demethylase system . Severe retinopathy of prematurity with retinal detachment in monozygotic twins . We report a monochorionic diamniotic twin pair born at 29 weeks of gestation in which both twins developed severe retinopathy of prematurity ( P04000 ) with retinal detachment . The pregnancy was terminated due to reversal of donor-recipient phenotypes in possible TTTS . Both twins had unstable cardiopulmonary status during the first week , and developed chronic lung disease . The larger twin , born at 1372 g , developed stage 4a P04000 in both eyes , and the smaller twin , born at 1168 g , developed stage 4a P04000 in the left eye . Genetic analysis of Q00604 , Q9ULV1 , O75197 , O95859 genes revealed no mutations ; however , P15692 gene polymorphism analysis showed heterozygous carrier state of the P15692 936T allele in both twins , which is a risk factor for threshold P04000 in Japanese newborn infants . We speculate the synergistic effects of unstable perinatal cardiopulmonary status and genetic predisposition due to P15692 936C > T polymorphism caused the development of severe P04000 with retinal detachment . P04035 inhibitor , atorvastatin , promotes sensorimotor recovery , suppressing acute inflammatory reaction after experimental intracerebral hemorrhage . BACKGROUND AND PURPOSE : Statins have neuroprotective effects on ischemic stroke . They modify the endothelial function , increase blood flow , and inhibit thrombus formation , which are independent of lipid-lowering effects . However , whether statins have a protective effect toward hemorrhagic stroke is yet unknown . To test this possibility , we attempted to determine the effect of atorvastatin on experimental intracerebral hemorrhage ( ICH ) . METHODS : ICH was induced using stereotaxic infusion of collagenase into the left basal ganglia in adult rats . DB01076 ( 1 mg/kg or 10 mg/kg ) or phosphate-buffered saline was administered for 2 weeks . To monitor the sensorimotor deficits , limb placing and Rotorod tests were performed . Hematoma volume , brain water content , and hemispheric atrophy were analyzed . Immunohistochemical staining for myeloperoxidase ( P05164 ) , microglia ( OX42 ) , inducible nitric oxide synthase ( P35228 ) , or endothelial nitric oxide synthase ( P29474 ) was performed . Perihematomal cell death was determined by TUNEL staining . RESULTS : The atorvastatin-treated ICH group showed better performance on Rotorod and limb placing tests when compared with the vehicle-treated group ( P < 0.01 ) . The hematoma volumes between groups were not different , but the brain water content and hemispheric atrophy were reduced in the atorvastatin-treated ICH group . DB01076 reduced TUNEL-positive cells , P35228 expression , and P05164 -positive or OX42-positive cells in the perihematomal regions in a dose-dependent manner , whereas it increased P29474 expression . CONCLUSIONS : The present study shows that atorvastatin reduces the perihematomal cell death via antiinflammation , which is associated with sensorimotor recovery after experimental ICH . [ Hemostatic evaluation of a patient with haloperidol-induced neuroleptic malignant syndrome associated with disseminated intravascular coagulation ] . A 94-year-old man who had been admitted to our hospital for the treatment of senile dementia and restless behavior exhibited consciousness disturbances , acute respiratory failure , high fever , and thrombocytopenia the day after receiving haloperidol as prescribed by a psychiatrist . On the fourth day following administration of haloperidol , acute renal failure with rhabdomyolysis and disseminated intravascular coagulation ( DIC ) developed in the patient , who was accordingly given a diagnosis of haloperidol-induced neuroleptic malignant syndrome ( Q5H8A3 ) associated with DIC . He was then given heparin and antithrombin III , and his DIC symptoms improved soon thereafter . Elevated plasma levels of tissue factor and tumor necrosis factor-alpha ( P01375 ) were sustained during this therapy course . Other cytokines , including interleukin P01584 , P60568 and P05231 , were not elevated . There are activation of extrinsic coagulation and an elevated level of P01375 during acute renal failure and rhabdomyolysis associated with Q5H8A3 , which is thought to trigger the onset of DIC . P10275 in Sertoli cells is not required for testosterone-induced suppression of spermatogenesis , but contributes to Sertoli cell organization in Utp14b jsd mice . DB00624 acting through the androgen receptor ( AR ) maintains the arrest of spermatogonial differentiation in juvenile spermatogonial depletion ( jsd mutation in the Utp14b gene ) mutant adult male mice . It is not known which of the somatic cell types expressing AR mediates this inhibition . To determine whether Sertoli cells are responsible , we selectively eliminated AR in Sertoli cells in jsd mice containing a floxed-Ar gene and an anti-Müllerian hormone-Cre transgene . In these Sertoli AR-knockout ( SCARKO ) -jsd mice , spermatogonial differentiation did not recover . However , the normal organization of Sertoli cell nuclei was drastically disrupted in SCARKO-jsd mice compared with SCARKO or jsd mice . In addition , the extent of ectoplasmic specializations was reduced ; tight junctions were not found ; vinculin , an anchoring protein found in ectoplasmic specializations , became uniformly distributed in the cytoplasm ; and the adult Sertoli cells showed excess heterochromatin subjacent to their nuclear envelope . Despite the abnormalities in Sertoli cells in SCARKO-jsd mice , global suppression of testosterone action and levels was still effective in restoring the differentiated germ cells , and this was accompanied by an improved arrangement of Sertoli cell nuclei . We conclude that Sertoli cells are not targets for the testosterone-mediated inhibition of spermatogonial differentiation in jsd mice , and that both AR in Sertoli cells and the presence of differentiated germ cells contribute to maintaining the organization of Sertoli cells within the seminiferous tubules . Preserved ex vivo inflammatory status in decidual cells from women with preterm labor and subclinical intrauterine infection . OBJECTIVE : To compare the inflammatory response preserved ex vivo by decidual cells isolated from women who experienced preterm labor with and without subclinical intrauterine infection . METHODS : Fetal membranes were obtained after cesarean section from 35 women who delivered before 37 weeks of gestation following spontaneous preterm labor , with no clinical evidence of intrauterine infection . Decidua was microbiologically tested and cultured . Concentrations of anti-inflammatory cytokines ( P60568 , P05112 , P22301 ) , pro-inflammatory cytokines ( P05231 , P10145 , IL-1β and P01375 -α ) , and matrix metalloproteinases ( P03956 , P08253 , P08254 , P09237 , P22894 , P14780 ) were measured in the supernatants using Bio-Plex , and prostaglandin E(2) ( PGE(2) ) was measured by enzyme immunoassay . RESULTS : Subclinical infection was confirmed in 10 women ( 28.5 % ) . Microorganisms isolated were Ureaplasma urealyticum ( 4 ) , group B streptococci ( 3 ) , Gardnerella vaginalis ( 1 ) , and Escherichia coli ( 2 ) . We found a significant increase of pro-inflammatory cytokines and a significant decrease of anti-inflammatory cytokines in supernatants from decidual cells obtained from women with preterm labor and subclinical intrauterine infection compared to women without infection . Secretion of P03956 , P22894 , P14780 and PGE(2) was significantly higher in infected women . Secretion of P10145 by decidual cells from infected women persisted upon repeated in vitro culture passages . CONCLUSIONS : Almost 30 % of idiopathic preterm labor cases were associated with subclinical intrauterine infection , and decidual cells isolated from these cases preserved an ex vivo inflammatory status after in vivo bacterial exposure . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . DB00125 -vasopressin directly promotes a thermogenic and pro-inflammatory adipokine expression profile in brown adipocytes . DB00125 -vasopressin ( AVP ) - via activation of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis - may play a role in the regulation of energy homeostasis and related cardiovascular complications . Brown adipose tissue ( Q14032 ) - via dissipation of energy in the form of heat - contributes to whole body energy balance . Q14032 expresses vasopressin receptors . We investigated direct effects of AVP on brown adipose endocrine and metabolic functions . P25874 -1 protein expression in differentiated brown adipocytes was induced after acute exposure of adipocytes to AVP . This effect was time-dependent with a maximum increase after 8h . AVP also induced a time- and dose-dependent increase in p38 Q96HU1 kinase phosphorylation . Pharmacological inhibition of p38 Q96HU1 kinase with SB 202190 abolished the induction of P25874 -1 protein expression . Furthermore , while acute AVP treatment enhanced mRNA expression of P13500 and P05231 , adiponectin mRNA expression was reduced . Yet , on the level of intracellular glucose uptake , there was no AVP-induced change of adipose insulin-induced glucose uptake . Finally , there was no difference in lipid accumulation between control and AVP-treated cells . Taken together , our data demonstrate direct effects of AVP on thermogenic , inflammatory , and glucoregulatory gene expression in brown adipocytes , thus expanding the hitherto known spectrum of this neuropeptides 's biological effects and suggesting a direct adipotropic role as a stress-promoting factor . P10275 signaling induced by supraphysiological doses of dihydrotestosterone in human peripheral blood lymphocytes . Anabolic androgenic steroids , a class of steroid hormones related to testosterone , are natural ligands of androgen receptor ( AR ) , a member of the nuclear receptor superfamily of ligand-activated transcription factors . AR binds specific DNA elements , known as androgen-response elements . DB00624 , the main male sexual hormone , binds AR directly and indirectly , through conversion into dihydrotestosterone ( DB02901 ) , its more active metabolite . Anabolic androgenic steroids are frequently detected in the urine of doped athletes ; their consumption is also growing among sport amateurs and adolescents . The effects of androgens can differ depending on the target cells and/or tissues . To gain insight into transcription activation mechanisms of AR , we investigated AR protein signaling in human peripheral blood lymphocytes treated with supraphysiological doses of DB02901 . We performed a comparative proteomic analysis and we identified about 30 differentially expressed proteins . At least five species contained a consensus androgen-response elements sequence in the promoter region of related coding genes . The analysis also revealed that high doses of DB02901 activate the drug detoxification process , could stimulate an increase in cell motility and exert a prosurvival effect rather than an apoptotic one . The aging suppressor klotho : a potential regulator of growth hormone secretion . Q9UEF7 is a transmembranal protein highly expressed in the kidneys , choroid plexus , and anterior pituitary . Q9UEF7 can also be cleaved and shed and acts as a circulating hormone . Q9UEF7 -deficient mice ( kl/kl mice ) develop a phenotype resembling early aging . Several lines of evidence suggest a role for klotho in the regulation of growth hormone ( GH ) secretion . The kl/kl mice are smaller compared with their wild-type counterparts , and their somatotropes show reduced numbers of secretory granules . Moreover , klotho is a potent inhibitor of the P05019 pathway , a negative regulator of GH secretion . Therefore , we hypothesized that klotho may enhance GH secretion . The effect of klotho on GH secretion was examined in GH3 rat somatotrophs , cultured rat pituitaries , and cultured human GH-secreting adenomas . In all three models , klotho treatment increased GH secretion . Prolonged treatment of mice with intraperitoneal klotho injections increased mRNA levels of P05019 and P05019 -binding protein-3 mRNA in the liver , reflecting increased serum GH levels . In accord with its ability to inhibit the P05019 pathway , klotho partially restored the inhibitory effect of P05019 on GH secretion . Q9UEF7 is known to be a positive regulator of basic P09038 signaling . We studied rat pituitaries and human adenoma cultures and noted that P09038 increased GH secretion and stimulated P27361 /2 phosphorylation . Both effects were augmented following treatment with klotho . Taken together , our data indicate for the first time that klotho is a positive regulator of GH secretion and suggest the P05019 and P09038 pathways as potential mediators of this effect . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . P10275 is widely expressed in bovine placentomes and up-regulated during differentiation of bovine trophoblast giant cells . OBJECTIVES : In cattle no biological role has been definitely identified for placental estrogens and progesterone . However , in the bovine trophoblast androgens may also be produced and have local effects . Thus , the aims of this study were to identify androgen receptor ( AR ) expressing cells and to monitor testosterone tissue concentrations in bovine placentomes throughout gestation . METHODS : Placental AR expression was characterized at the mRNA and protein level applying conventional and real-time RT-qPCR , western blot and immunohistochemistry . DB00624 was measured by radioimmunoassay . RESULTS : AR-mRNA was qualitatively detected from day 50 of gestation until term . Mean relative gene expression levels were constant between day 100 and late gestation . A slight non-significant increase was observed in the prepartal period . With immunohistochemistry distinct nuclear signals were predominantly observed in invasive trophoblast giant cells ( P21980 ) from day 80 until term . In mature P21980 of the trophoblast , immature P21980 and uninucleated trophoblast cells , stromal cells of the chorionic villi , caruncular epithelial and stromal cells immunoreactive score values were low at early and midgestation but increased significantly ( p < 0.01 ) during late gestation and remained high until parturition . With western blot in placentomal tissue a specific band of approximately 110 kDa was detected as it was the case in epididymis used as a positive control . DB00624 concentrations increased from 0.70 ± 0.29 pmol/g wet tissue between days 60-220 to 4.22 ± 1.29 pmol/g during late gestation ( p < 0.001 ) . DISCUSSION : The results are consistent with androgens as active products of bovine placental steroidogenesis . The substantial up-regulation of AR expression during P21980 differentiation suggests that androgens may be related to this process . P10275 (CAG)n polymorphism and androgen levels in women with systemic lupus erythematosus and healthy controls . Systemic lupus erythematosus ( SLE ) is an autoimmune disorder that affects mainly females . Therefore , interrelations between the reproductive and immune system have been assumed . Considering the complex influence of hormones and receptors , we aimed to investigate the influence of androgens and androgen receptor ( AR ) polymorphism in women with SLE . One hundred and sixteen patients and 44 healthy women were investigated . DB00624 , sex hormone-binding globulin ( P04278 ) , dehydroepiandrosterone-sulphate ( DHEAS ) concentrations and AR (CAG)n polymorphism were determined . SLE patients had significantly lower levels of total and free testosterone and DHEAS in comparison with the controls . No differences in the CAG repeat length between the groups were established . Women with two alleles carrying more than 22 CAG repeats had significantly higher levels of P04278 ( 101.51 ± 61.81 vs. 69.22 ± 45.93 nmol/l , p = 0.015 ) and DHEAS ( 3.11 ± 2.65 vs. 2.11 ± 3.06 μmol/l , p = 0.007 ) and a tendency to higher testosterone concentrations ( 2.35 ± 2.10 vs. 1.71 ± 1.70 nmol/l , p = 0.056 ) in comparison with other women . The CAG repeat length in the relatively longer (CAG)n allele was inversely related to the Systemic Lupus International Collaborating Clinics/ P10323 index ( r = -0.258 , p = 0.009 ) . In conclusion , the androgen receptor (CAG)n polymorphism is not related to the development of SLE , but it could modulate the severity of the lupus chronic damages as well as the androgen levels in women . P10275 targets NFkappaB and TSP1 to suppress prostate tumor growth in vivo . The androgen role in the maintenance of prostate epithelium is subject to conflicting opinions . While androgen ablation drives the regression of normal and cancerous prostate , testosterone may cause both proliferation and apoptosis . Several investigators note decreased proliferation and stronger response to chemotherapy of the prostate cancer cells stably expressing androgen receptor ( AR ) , however no mechanistic explanation was offered . In this paper we demonstrate in vivo anti-tumor effect of the AR on prostate cancer growth and identify its molecular mediators . We analyzed the effect of AR on the tumorigenicity of prostate cancer cells . Unexpectedly , the AR-expressing cells formed tumors in male mice at a much lower rate than the AR-negative controls . Moreover , the AR-expressing tumors showed decreased vascularity and massive apoptosis . AR expression lowered the angiogenic potential of cancer cells , by increasing secretion of an anti-angiogenic protein , thrombospondin-1 . AR activation caused a decrease in RelA , a subunit of the pro-survival transcription factor NFkappaB , reduced its nuclear localization and transcriptional activity . This , in turn , diminished the expression of its anti-apoptotic targets , Bcl-2 and P05231 . Increased apoptosis within AR-expressing tumors was likely due to the NFkappaB suppression , since it was restricted to the cells lacking nuclear ( active ) NFkappaB . Thus we for the first time identified combined decrease of NFkappaB and increased TSP1 as molecular events underlying the AR anti-tumor activity in vivo . Our data indicate that intermittent androgen ablation is preferable to continuous withdrawal , a standard treatment for early-stage prostate cancer . ( c ) 2007 Wiley-Liss , Inc . DB00624 inhibits matrix metalloproteinase-1 production in human endometrial stromal cells in vitro . P10275 ( AR ) is reported to be expressed in human uterine endometrium , but not much information is available on the role of androgens in human endometrium . The purpose of this study is to investigate the role of androgens in the regulation of matrix metalloproteinase ( MMP ) -1 , which is one of the important MMPs for menstruation and embryo implantation in human endometrium . Human endometrial stromal cells ( HESCs ) were obtained from human endometrium by enzymatic dissociation method . Purified HESCs were incubated with 17beta-estradiol ( E2 ) , testosterone , or E2 + testosterone . Progestins ( natural progesterone or medroxyprogesterone acetate ) or vehicle ( dimethyl sulfoxide ) were also added to the media instead of testosterone . Furthermore , hydroxyflutamide (FLU),a specific AR antagonist , was also supplemented to cultured media . The amounts of P03956 in cultured media and in HESC lysates were examined by ELISA measurements and western blotting analysis respectively . The expression of ARmRNA in HESCs RNA was analyzed by RT-PCR . DB00624 significantly inhibited P03956 in both cultured media and cell lysates in a dose-dependent manner . Progestins also inhibited P03956 . Furthermore , FLU completely recovered the decrease of P03956 induced by testosterone . ARmRNA was detected in all HESCs RNA . The present study demonstrated that the secretion and production of P03956 in HESCs in vitro were inhibited by testosterone through androgen receptors in a manner similar to that seen for progesterone . These findings indicate that androgen may play an important role in morphological and functional changes of human endometrium . P10275 CAG repeat polymorphism is associated with serum testosterone levels , obesity and serum leptin in men with type 2 diabetes . OBJECTIVE : To determine the relationships between androgen receptor CAG repeat polymorphism length ( AR CAG ) , sex hormones and clinical variables in men with type 2 diabetes ( DM2 ) . Men with DM2 are known to have a high prevalence of low testosterone levels . Studies suggest that testosterone replacement therapy may improve insulin sensitivity and glycaemic control in men with DM2 and reduces central obesity and serum leptin . AR CAG is known to correlate negatively with AR sensitivity and positively with body fat , insulin levels , and leptin in healthy men . DESIGN : Cross-sectional study set in a district general hospital diabetes centre . METHODS : Sex hormones , AR CAG and symptoms of hypogonadism were assessed in 233 men with DM2 . Associations were sought between these variables and others such as obesity , leptin , glycaemic control , and blood pressure . RESULTS : DB00624 was negatively associated and AR CAG positively associated with obesity and leptin . The associations of AR CAG with leptin and obesity were independent of testosterone , estradiol , gonadotropins , and age . AR CAG was also independently associated with total , bioavailable and free testosterone , LH , waist circumference , body mass index , leptin , and systolic blood pressure . There was no association of AR CAG with sex hormone binding globulin , estradiol , HbA(1C) or the symptoms of hypogonadism . CONCLUSIONS : The association of longer AR CAG with obesity and leptin suggests that shorter AR CAG may have an influence in maintaining healthy anthropomorphics and metabolism in men with DM2 . DB00624 and LH levels are higher in men with longer AR CAG , probably reflecting reduced negative feedback through a less sensitive receptor . Delayed haemolytic transfusion reaction caused by anti-M antibody in a patient receiving interleukin-2 and interferon for metastatic renal cell cancer . Anti-M is usually a naturally occurring cold-reactive immunoglobulin M ( IgM ) antibody , often with an immunoglobulin G ( IgG ) component , and is seldom implicated in delayed haemolytic transfusion reactions ( P10275 ) . However , cases have been reported . In the majority , a P10275 is not suspected until further blood is requested and a new antibody is detected on pretransfusion testing . We describe the case of a young man receiving therapy with interleukin-2 ( P60568 ) and interferon-alpha ( IFN-alpha ) for metastatic renal cell cancer who developed a clinically suspected P10275 that was confirmed serologically to be caused by anti-M , reactive at 37 degrees C . We discuss the possible role of his biochemotherapy in the development of the P10275 . Cholinergic receptor up-regulates P35354 expression and prostaglandin E(2) production in colon cancer cells . The M(3) muscarinic cholinergic receptor has important physiological functions on normal colonic cells . It is frequently expressed on human colon cancer cells and is biologically active . Although it is mitogenic in certain cell models , the importance of this receptor on colon carcinogenesis is unknown . In the present study we have determined expression of the M(3) receptor on human colon cancer tissue compared with matched normal tissue and examined the downstream effect of receptor activation in the HT-29 human colon carcinoma cell line . Using reverse transcription-PCR , M(3) receptor RNA expression was detected in all matched colon carcinoma and normal specimens from eight patients . Five of the eight ( 62 % ) patients showed an up to 8-fold greater level of M(3) receptor expression in cancer compared with the matched normal tissue . Exposure of HT-29 cells to carbachol , a stable receptor agonist , results in a 10-fold increase in cyclooxygenase-2 ( P35354 ) protein . This induction of P35354 protein was dose dependent and was inhibited by the cholinergic receptor antagonist N-methylscopolamine ( Q5H8A3 ) . DB00411 caused a dose-dependent increase in prostaglandin E(2) ( PGE(2) ) , the main product of cyclooxygenase activity . The maximum stimulatory effect ( 40-fold increase ) was noted with 1mM carbachol . The increase in PGE(2) was completely abolished by Q5H8A3 and by the P35354 selective inhibitor NS398 . This suggests that the M(3) receptor mediates PGE(2) production by a mechanism involving P35354 . As P35354 and PGE(2) are known promoters of gastrointestinal cancer , these data suggest that M(3) receptor activation may facilitate progression of colon carcinoma , in part by a P35354 -mediated cellular mechanism . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P10275 signalling in peritubular myoid cells is essential for normal differentiation and function of adult Leydig cells . DB00624 synthesis depends on normal Leydig cell ( LC ) development , but the mechanisms controlling this development remain unclear . We recently demonstrated that androgen receptor ( AR ) ablation from a proportion of testicular peritubular myoid cells ( PTM-ARKO ) did not affect LC number , but resulted in compensated LC failure . The current study extends these investigations , demonstrating that PTM AR signalling is important for normal development , ultrastructure and function of adult LCs . Notably , mRNAs for LC markers [ e.g. steroidogenic factor 1 ( Nr5a1 ) , insulin-like growth factor ( Igf-1 ) and insulin-like factor 3 ( Insl3 ) ] were significantly reduced in adult PTM-ARKOs , but not all LCs were similarly affected . Two LC sub-populations were identified , one apparently ' normal ' sub-population that expressed adult LC markers and steroidogenic enzymes as in controls , and another ' abnormal ' sub-population that had arrested development and only weakly expressed P51460 , luteinizing hormone receptor , and several steroidogenic enzymes . Furthermore , unlike ' normal ' LCs in PTM-ARKOs , the ' abnormal ' LCs did not involute as expected in response to exogenous testosterone . Differential function of these LC sub-populations is likely to mean that the ' normal ' LCs work harder to compensate for the ' abnormal ' LCs to maintain normal serum testosterone . These findings reveal new paracrine mechanisms underlying adult LC development , which can be further investigated using PTM-ARKOs . P10275 in human skin cytosol . Human skin , an accessible tissue , is an androgen target organ . We have measured the androgen-binding capacity of human skin cytosol using either 5 alpha[3H]dihydrotestosterone ( [3H] DB02901 ) or [3H]methyltrienolone ( [3H]R-1881 ) as ligand . Samples were incubated for 20 h at 0 C , and dextran-coated charcoal was used to separate bound from free steroids . The androgen receptor has a high affinity for both ligands ( 0.23 +/- 0.04 nM for [3H] DB02901 ; 0.32 +/- 0.16 nM for [3H]R-1881 ) . DB00624 , cyproterone acetate , and , to a lesser extent , estradiol also bind this protein . Progesterone displaces R-1881 from its binding sites , whereas its 5 alpha-reduced metabolite somewhat inhibits DB02901 binding . The highest binding capacity is measured in cytosol of skin from external genitalia ( 129.14 +/- 58.0 fmol/g skin ; n = 34 ) ; it is lower in pubic skin ( 21.8 +/- 13 fmol/g skin ; n = 6 ) . There is no variation as a function of age or sex in genital skin ; the higher concentrations observed in the cytosol of pubic skin of women compared to that of men are probably related to lower levels of endogenous steroids . Whereas most patients with the complete form of the testicular feminization syndrome do not have detectable concentrations of androgen receptor , one patient with apparent complete clinical androgen insensitivity had a normal androgen-binding capacity . The parity of values in genital skin from men and women , the absence of variation with age , and the presence of a cytosolic androgen receptor in some androgen-insensitive patients suggest that the androgen receptor in human skin cytosol is not regulated by androgens . P10275 -dependent transactivation of growth arrest-specific gene 6 mediates inhibitory effects of testosterone on vascular calcification . Recent epidemiological studies have found that androgen deficiency is associated with a higher incidence of cardiovascular disease in men . However , little is known about the mechanism underlying the cardioprotective effects of androgens . Here we show the inhibitory effects of testosterone on vascular calcification and a critical role of androgen receptor ( AR ) -dependent transactivation of growth arrest-specific gene 6 ( Gas6 ) , a key regulator of inorganic phosphate ( P(i) ) -induced calcification of vascular smooth muscle cells ( VSMC ) . DB00624 and nonaromatizable androgen dihydrotestosterone inhibited P(i)-induced calcification of human aortic VSMC in a concentration-dependent manner . Androgen inhibited P(i)-induced VSMC apoptosis , an essential process for VSMC calcification . The effects on VSMC calcification were mediated by restoration of P(i)-induced down-regulation of Gas6 expression and a subsequent reduction of Akt phosphorylation . These effects of androgen were blocked by an AR antagonist , flutamide , but not by an estrogen receptor antagonist , DB00947 . We then explored the mechanistic role of the AR in Gas6 expression and found an abundant expression of AR predominantly in the nucleus of VSMC and two consensus ARE sequences in the Gas6 promoter region . DB02901 stimulated Gas6 promoter activity , and this effect was abrogated by flutamide and by AR siRNA . Site-specific mutation revealed that the proximal ARE was essential for androgen-dependent transactivation of Gas6 . Furthermore , chromatin immunoprecipitation assays demonstrated ligand-dependent binding of the AR to the proximal ARE of Gas6 . These results indicate that AR signaling directly regulates Gas6 transcription , which leads to inhibition of vascular calcification , and provides a mechanistic insight into the cardioprotective action of androgens . Androgen receptors . DB00624 and its active metabolite dihydrotestosterone exert their influence on target cells through a specific intracellular protein receptor . Structural abnormalities of this receptor lead to a diminished androgen action within the cell and result in the syndrome of androgen insensitivity . Androgen insensitivity is classified on the basis of whether the insensitivity is complete or partial and whether the androgen receptor is normally present ( AR(+) ) , absent ( AR(-) ) or diminished ( AR(+/-) ) . All patients with androgen insensitivity have normal or high plasma levels of testosterone and elevated serum LH . Patients with complete androgen insensitivity are phenotypically female . The clinical presentation of partial androgen insensitivity is variable , ranging from a minimal amount of virilization to a completely masculine appearance . All patients described with a syndrome of androgen insensitivity are infertile . The influence of androgen receptor function in the pathogenesis of benign prostatic hypertrophy is being investigated . P10275 content is also being studied as a possible marker of responsiveness to hormonal therapy in prostatic carcinoma . P04035 inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection . P10275 up-regulates insulin-like growth factor binding protein-5 ( P24593 ) expression in a human prostate cancer xenograft . The insulin-like growth factor ( IGF ) binding proteins ( IGFBPs ) are important modulators of IGF action in many tissues including human prostate . IGFBPs and the androgen receptor ( AR ) are expressed in CWR22 , an androgen-dependent epithelial cell human CaP xenograft that retains biological characteristics of human CaPs , including regression following androgen withdrawal and recurrent growth of AR-containing cells in the absence of testicular androgens beginning several months after castration . Northern blot and in situ hybridization analyses demonstrated that P24593 is androgen-regulated in CWR22 . P24593 messenger RNA ( mRNA ) decreased by 90 % following castration of tumor-bearing mice compared with noncastrate androgen-stimulated mice . DB00624 treatment of CWR22 tumor-bearing mice 6 or 12 days after castration increased P24593 mRNA 10- to 12-fold . Levels of other IGFBP mRNAs did not change following androgen withdrawal and replacement . P24593 protein in tumor extracts bound 125I-labeled P05019 in ligand blot assays and the amounts of P24593 measured by immunoblotting paralleled the levels of P24593 mRNA . Androgen-induced expression of P24593 was at a maximum level within 24 h after testosterone replacement , whereas the major increase in cell proliferation as measured by Ki-67 immunostaining occurred between 24-48 h . This time course suggested P24593 may be a mediator of androgen-induced growth of CWR22 . In tumors that recurred several months following castration , P24593 mRNA and protein increased to levels that approached those in androgen-stimulated CWR22 tumors from noncastrate mice . P24593 immunohistochemical staining of prostate tissue specimens from patients was stronger in androgen-dependent and androgen-independent CaP than in areas of intraepithelial neoplasia ( P63167 ) or benign prostatic hyperplasia ( BPH ) . P24593 mRNA in these specimens was localized predominantly to stromal cells and P24593 protein to epithelial cell membranes . Chemical ablation of androgen receptor in prostate cancer cells by the histone deacetylase inhibitor LAQ824 . P10275 plays a critical role in the development of primary as well as advanced hormone-refractory prostate cancer . Therefore , ablation of androgen receptor from prostate cancer cells is an interesting concept for developing a new therapy not only for androgen-dependent prostate cancer but also for metastatic hormone-refractory prostate cancer , for which there is no effective treatment available . We report here that LAQ824 , a cinnamyl hydroxamatic acid histone deacetylase inhibitor currently in human clinical trials , effectively depleted androgen receptor in prostate cancer cells at nanomolar concentrations . LAQ824 seemed capable of depleting both the mutant and wild-type androgen receptors in either androgen-dependent and androgen-independent prostate cancer cells . Although LAQ824 may exert its effect through multiple mechanisms , several lines of evidence suggest that inactivation of the heat shock protein-90 ( Hsp90 ) molecular chaperone is involved in LAQ824-induced androgen receptor depletion . Besides androgen receptor , LAQ824 reduced the level of Hsp90 client proteins HER-2 ( ErbB2 ) , Akt/ P31749 , and P04049 in LNCaP cells . Another Hsp90 inhibitor , 17-allyamino-17-demethoxygeldanamycin ( 17- P29372 ) , also induced androgen receptor diminution . LAQ824 induced Hsp90 acetylation in LNCaP cells , which resulted in inhibition of its DB00171 -binding activity , dissociation of Hsp90-androgen receptor complex , and proteasome-mediated degradation of androgen receptor . Consequently , LAQ824 blocked androgen-induced prostate-specific antigen production in LNCaP cells . LAQ824 effectively inhibited cell proliferation and induced apoptosis of these prostate cancer cells . These results reveal that LAQ824 is a potent agent for depletion of androgen receptor and a potential new drug for prostate cancer . Serum testosterone plays an important role in the metastatic ability of castration resistant prostate cancer . PURPOSE : Prostate cells are dependent on androgens for growth and proliferation . Androgen deprivation therapy is the recommended treatment for advanced/metastatic prostate cancer . Under this therapy , prostate cancer will inevitably progress to castration resistant prostate cancer ( CRPC ) . Despite putative castration resistance , testosterone might still play a crucial role in the progression of CRPC . The goal of this study was to determine the role of testosterone in the formation of metastases of CRPC in both in vitro and in vivo settings . METHODS : In vitro , the effect of testosterone and the non-aromatizable androgen methyltrienolone on migration , invasion and proliferation of a castration-resistant prostate cancer rat cell line ( Dunning R3327-MATLyLu ) was assessed using a transwell assay and a sulforhodamine B assay and immunohistochemical detection of ki67 . P10275 status was determined using Western blot . In vivo , Copenhagen rats were divided in four groups ( males , females , castrated males and females with testosterone suppletion ) and inoculated with MATLyLu cells . Tumor size was assessed daily . RESULTS : DB00624 increased cell migration and invasion in a concentration-dependent manner in vitro . DB00624 did not affect in vitro cell proliferation . No difference was shown between the effect of testosterone and methyltrienolone . In vivo , in groups with higher levels of circulating testosterone , more rats had (micro)metastases compared with groups with low levels of testosterone . No effect was observed on primary tumor size/growth . CONCLUSIONS : Despite assumed castration resistance , progression of prostate cancer is still influenced by androgens . Therefore , continuous suppression of serum testosterone in patients who show disease progression during castration therapy is still warranted . DB00624 stimulates proliferation and inhibits interleukin-6 production of normal and hereditary gingival fibromatosis fibroblasts . Hereditary gingival fibromatosis ( P14210 ) is a rare oral condition characterized by a slow and progressive enlargement of the gingiva , involving both the maxilla and mandible . In vitro , P14210 fibroblasts demonstrate a proliferative index significantly higher than fibroblasts from normal gingiva ( NG ) . The objective of this study was to determine the effect of dihydrotestosterone on the proliferation of gingival fibroblasts derived from patients with P14210 ( n = 4 ) and from four healthy individuals . Additionally , we analyzed the effect of dihydrotestosterone on interleukin-6 ( P05231 ) production and determined the expression levels of androgen receptors in NG and P14210 fibroblasts . Gingival fibroblasts from NG and P14210 were incubated with increasing concentrations of dihydrotestosterone with or without androgen blockers , and cultured for 24 h , and the proliferation index was determined by automated cell counter . P05231 production , in this system , was quantified using a " capture " enzyme-linked immunosorbent assay ( ELISA ) . Semi-quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR ) was performed to measure the mRNA expression of androgen receptors . The results indicated that dihydrotestosterone simultaneously downregulates the production of P05231 and upregulates the cell proliferation . DB01216 and cyprosterone acetate , two anti-androgens , partially reversed these effects . P10275 mRNA expression was identified in both NG and P14210 fibroblasts ; however , the levels in NG were higher than those observed in P14210 . These results show that testosterone coordinates the proliferation and production of P05231 of normal and P14210 fibroblasts . Axotomy transiently down-regulates androgen receptors in motoneurons of the spinal nucleus of the bulbocavernosus . DB00624 is an important trophic factor for motoneurons in the spinal nucleus of the bulbocavernosus ( SNB ) , and SNB motoneurons are more responsive to testosterone than are other motoneurons . Axonal injury during early postnatal life prevents the normal development of steroid-sensitivity by adult SNB motoneurons . Axonal injury also causes changes in the expression by motoneurons of a wide range of proteins , including the up-regulation of trophic factor receptors . We have used a polyclonal antibody ( PG-21 ; G.S. Prins ) to study the expression of androgen receptors in SNB motoneurons after axonal injury . PG-21 labeled motoneuronal nuclei in the lower lumbar spinal cord of rats in a pattern that matched autoradiographic reports of androgen accumulation in this region of the nervous system . A population of numerous , small cells located dorsal to the central canal also showed evidence of androgen receptor expression . Cutting the axons of SNB motoneurons in adulthood or in development caused a decrease in androgen receptor immunoreactivity in SNB motoneurons . This is the first report that a trophic factor receptor in motoneurons is down-regulated after axonal injury , and is interesting in light of reports that testosterone treatment can facilitate motoneuronal regeneration after nerve cut . P10275 levels subsequently returned to normal , regardless of the age at axotomy , providing no evidence for a lasting effect of developmental axotomy on androgen receptor levels in SNB motoneurons . Thus , axotomy-induced down-regulation of androgen receptors does not underlie the inability of SNB motoneurons to respond to androgen treatment several months after pudendal nerve cut in development . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . P10275 -immunoreactive cells in ram hypothalamus : distribution and co-localization patterns with gonadotropin-releasing hormone , somatostatin and tyrosine hydroxylase . DB00624 exerts important feedback effects on the hypothalamus of the ram to influence reproductive functioning . To provide a neuroanatomical basis for understanding this androgen action , the present study has examined androgen receptor ( AR ) immunoreactivity within the hypothalamus and adjacent brain areas of the intact non-breeding season ram . The largest populations of AR-immunoreactive cells were detected in the medial preoptic area , infundibular and premammillary nuclei in addition to the ventromedial nucleus ( VMN ) where cells were found distributed throughout its medial and lateral divisions . Smaller numbers of AR-expressing cells were identified in the bed nucleus of the stria terminalis and anterior hypothalamic area ( DB00551 ) including the paraventricular , but not the supraoptic , nucleus . Double-labelling immunocytochemistry revealed the presence of AR immunoreactivity in only 2 of 460 gonadotropin-releasing hormone ( DB00644 ) neurons . A very small population of TH-immunoreactive cells located in the lateral aspect of the DB00551 was found to contain ARs . Dopaminergic cells elsewhere in the hypothalamus , including the infundibular nucleus , did not display AR immunoreactivity . Nearly 50 % of AR-expressing cells in the lateral VMN were immunoreactive for somatostatin while less than 5 % of periventricular somatostatin neurons displayed AR immunoreactivity . These results show where ARs are expressed in the ram hypothalamus and indicate the neuroanatomical sites at which androgen may act to influence reproductive function . The absence of ARs in the neuroendocrine DB00644 and tuberoinfundibular dopaminergic cells suggests that androgens do not influence the genome of these cells in any direct manner . In contrast , the somatostatin neurons of the VMN appear to be an important target for circulating androgens in the non-breeding season ram . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB00624 potentiates the hypoxic ventilatory response of adult male rats subjected to neonatal stress . Neonatal stress disrupts development of homeostatic systems . During adulthood , male rats subjected to neonatal maternal separation ( Q5H8A3 ) are hypertensive and show a larger hypoxic ventilatory response ( HVR ) , with greater respiratory instability during sleep . Neonatal stress also affects sex hormone secretion ; hypoxia increases circulating testosterone of Q5H8A3 ( but not control ) male rats . Given that these effects of Q5H8A3 are not observed in females , we tested the hypothesis that testosterone elevation is necessary for the stress-related increase of the HVR in adult male rats . Pups subjected to Q5H8A3 were placed in an incubator for 3 h per day from postnatal day 3 to 12 . Control pups remained undisturbed . Rats were reared until adulthood , and the HVR was measured by plethysmography ( fractional inspired O2 = 0.12 , for 20 min ) . We used gonadectomy to evaluate the effects of reducing testosterone on the HVR . Gonadectomy had no effect on the HVR of control animals but reduced that of Q5H8A3 animals below control levels . Immunohistochemistry was used to quantify androgen receptors in brainstem areas involved in the HVR . P10275 expression was generally greater in Q5H8A3 rats than in control rats ; the most significant increase was noted in the caudal region of the nucleus tractus solitarii . We conclude that the abnormal regulation of testosterone is important in stress-related augmentation of the HVR . The greater number of androgen receptors within the brainstem may explain why Q5H8A3 rats are more sensitive to testosterone withdrawal . Based on the similarities of the cardiorespiratory phenotype of Q5H8A3 rats and patients suffering from sleep-disordered breathing , these results provide new insight into its pathophysiology , especially sex-based differences in its prevalence . Increased P05231 levels in pituitary-deficient patients are independently related to their carotid intima-media thickness . OBJECTIVE : Increased cardiovascular morbidity and mortality has been observed in patients with pituitary deficiency and untreated growth hormone deficiency ( GHD ) . We investigated peripheral inflammatory and fibrinolytic markers and their associations with arterial intima-media thickness ( IMT ) in GHD . DESIGN : Cross-sectional study . PATIENTS : Thirty-four patients with GHD , but without cardiovascular disease , were compared to healthy controls matched for age , sex , body mass index ( BMI ) and smoking habits . MEASUREMENTS : IMT of the common carotid artery , P02741 ( CRP ) , interleukin-6 ( P05231 ) , fibrinogen , plasminogen activator inhibitor-1 ( P05121 ) activity and tissue plasminogen activator antigen ( tPA-ag ) were measured . RESULTS : Median P05231 concentrations were increased by 208 % and 248 % in GHD patients compared to BMI-matched and nonobese controls , respectively . Median CRP and tPA-ag levels were increased by 237 % and 167 % in patients compared to nonobese controls , but not significantly different compared to BMI-matched controls . Plasma levels of fibrinogen and P05121 activity did not differ between groups . Age , low-density lipoprotein ( LDL ) cholesterol , tPA-ag and P05231 were positively correlated , and P05019 was negatively correlated to IMT in the patient group , but only age and P05231 were independently related to IMT . CONCLUSIONS : P05231 concentrations were increased in GHD patients compared to controls and independently related to IMT in patients . This finding may help to explain the variance in IMT and the increased vascular morbidity and mortality in hypopituitary patients with GHD . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Priming effects of macrophage colony-stimulating factor on monocytic leukemia cells in combination with chemotherapy : induction of programmed cell death in vivo . Two elderly patients with chronic myelomonocytic leukemia were treated with cytosine arabinoside ( DB00987 ) and aclarubicin ( P10323 ) under simultaneous administrations of macrophage colony-stimulating factor ( P09603 ) ( P62158 ) , and both obtained good responses . Examination of apoptosis using flow cytometry revealed induction of apoptotic death of leukemia cells by P62158 in Patient 2 , while neither induction of apoptotic death of leukemia cells nor clinical response were seen with CAG ( DB00987 , P10323 , and granulocyte colony-stimulating factor ) given prior to P62158 in Patient 1 . These findings suggested that chemotherapy combined with simultaneous administration of P09603 could effectively reduce monocytic leukemia cells by inducing programmed cell death . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Genes controlling spread of breast cancer to lung " gang of 4 " . Cancer-related mortality is caused in a large part by the metastasis of primary tumor . Each cancer has a particular way of spreading cancerous cells . Recently , genetic and pharmacological analysis identified the set of genes , such as epidermal growth factor receptor ligand epiregulin ( O14944 ) , cyclooxygenase-2 ( P35354 ) and matrix metalloproteinases 1 and 2 ( P03956 and P08253 ) that have been found to be associated with metastasis of breast cancer to lung . Inhibition of P00533 and P35354 could minimize lung metastasis of breast cancer in a clinical setting . In this review , we summarized the current knowledge on O14944 , P35354 , P03956 and P08253 in tumor development and metastasis . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . DB00624 modulation of dendritic spines of somatosensory cortical pyramidal neurons . Brain structures and functions are increasingly recognized to be directly affected by gonadal hormones , which classically determine reproductive functions and sexual phenotypes . In this regard , we found recently that ovariectomy trimmed the dendritic spines of female rat primary somatosensory cortical neurons and estradiol supplement reversed it . Here , we investigated whether in the male androgen also has a cortical modulatory effect . The dendritic arbors and spines of rat somatosensory cortical pyramidal neurons were studied following intracellular dye injection and three-dimensional reconstruction . Dendritic spines , but not length , of the layers III and V pyramidal neurons were found reduced at 2 weeks and rebounded slightly at 4 weeks and further at 8 and 24 weeks following castration , which , however , remained significantly fewer than those of the intact animals . Two weeks of osmotic pump-delivered testosterone treatment to animals castrated for 4 weeks replenished serum testosterone and reversed the densities of dendritic spines on these neurons to control animal levels . P10275 appears to mediate this effect as its antagonist flutamide reduced the dendritic spines of normal adult rats while causing a mild feedback surge of serum testosterone . On the other hand , blocking the conversion of testosterone to estrogen with the aromatase inhibitor anastrozole failed to alter the dendritic spine densities in male adult rats . In conclusion , these results support our hypothesis that testosterone acts directly on the androgen receptor in males to modulate the dendritic spines of somatosensory cortical output neurons . Recent androgen receptor antagonists in prostate cancer . P10275 has been shown to promote prostate cell growth and carcinogenesis of prostate cancer by up-regulating its target genes . DB00624 and dihydrotestosterone are two major hormones which bind to and activate androgen receptor . Targeting both the androgen receptor and the enzymes catalyzing the biosynthesis of testosterone and dihydrotestosterone has been shown to be clinically beneficial in the treatment of prostate cancer . Prostate cancer can become castration-resistant after long term treatment with chemo drugs , so efforts in finding compounds with improved efficiency to castration-resistant prostate cancer are urgently needed . In this review we summarized the studies on recent progress in the development of small molecular AR antagonists for the treatment of prostate cancer . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . P04637 mutation , epithelial-mesenchymal transition , and stemlike features in breast cancer subtypes . Altered p53 protein is prevalently associated with the pathologic class of triple-negative breast cancers and loss of p53 function has recently been linked to the induction of an epithelial-mesenchymal transition ( EMT ) and acquisition of stemness properties . We explored the association between P04637 mutational status and expression of some genes involved in the canonical TGF-β signaling pathway ( the most potent EMT inducer ) and in two early EMT associated events : loss of cell polarity and acquisition of stemness-associated features . We used a publicly accessible microarray dataset consisting of 251 p53-sequenced primary breast cancers . Statistical analysis indicated that mutant p53 tumors ( especially those harboring a severe mutation ) were consistent with the aggressive class of triple-negative cancers and that , differently from cell cultures , surgical tumors underexpressed some TGF-β related transcription factors known as involved in EMT ( P41134 , P47928 , P84022 , Q13485 , Q99717 , P37275 ) . These unexpected findings suggest an interesting relationship between p53 mutation , mammary cell dedifferentiation , and the concomitant acquisition of stemlike properties ( as indicated by the overexpression of O43490 and P46531 genes ) , which improve tumor cells aggressiveness as indicated by the overexpression of genes associated with cell proliferation ( P11802 , Q00534 , P46013 ) and migration ( P61073 , P03956 ) . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Androgen receptors , serum androgen levels and survival of breast cancer patients . Steroid receptor levels and serum androgen levels were determined in 61 breast cancer patients and 34 patients with non-malignant breast lesions . DB00624 and dehydroepiandrosterone-sulfate did not and androstenedione did show a difference between the two groups . Androgen levels had no influence on survival rates . P10275 ( AR ) levels correlated with progesterone receptor levels , but not with estrogen receptor levels or with tumor stage . Patients with positive AR findings had a better survival rate ; this was independent of tumor stage . AR findings may therefore be a prognostic index in breast cancer patients . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . Nursing supports neonatal porcine testicular development . The lactocrine hypothesis suggests a mechanism whereby milk-borne bioactive factors delivered to nursing offspring affect development of neonatal tissues . The objective of this study was to assess whether nursing affects testicular development in neonatal boars as reflected by : ( 1 ) Sertoli cell number and proliferation measured by GATA-4 expression and proliferating cell nuclear antigen immunostaining patterns ; ( 2 ) Leydig cell development and steroidogenic activity as reflected by insulin-like factor 3 ( P51460 ) , and P450 side chain cleavage ( scc ) enzyme expression ; and ( 3 ) expression of estrogen receptor-alpha ( P03372 ) , vascular endothelial growth factor ( P15692 ) A , and relaxin family peptide receptor ( RXFP ) 1 . At birth , boars were randomly assigned ( n = 6-7/group ) to nurse ad libitum or to be pan fed porcine milk replacer for 48 h . Testes were collected from boars at birth , before nursing and from nursed and replacer-fed boars at 50 h on postnatal day ( P01160 ) 2 . Sertoli cell proliferating cell nuclear antigen labeling index increased ( P < 0.01 ) from birth to P01160 2 in nursed , but not in replacer-fed boars . Sertoli cell number and testicular GATA-4 protein levels increased ( P < 0.01 ) from P01160 0 to P01160 2 only in nursed boars . Neither age nor nursing affected testicular P51460 , P450scc , P03372 , or P15692 levels . However , testicular relaxin family peptide receptor 1 ( Q9HBX9 ) levels increased ( P < 0.01 ) with age and were greater in replacer-fed boars on P01160 2 . Results suggest that nursing supports neonatal porcine testicular development and provide additional evidence for the importance of lactocrine signaling in pigs . Gene expression in sexually dimorphic muscles in sheep . DB00624 is known to act differentially on skeletal muscle from different regions of the body . Two genes likely to mediate the testosterone effect are insulin-like growth factor I ( P05019 ) , an important growth regulator acting in an autocrine and paracrine way , and androgen receptor ( AR ) , because receptor density could account for differential muscle growth . Another muscle-specific gene that may play a role in differential muscle growth is myostatin , a member of the transforming growth factor-beta superfamily , shown to be a negative regulator of skeletal muscle mass . The objective of this study was to quantify and compare the steady state expression of these three genes in two different skeletal muscles in sheep . Eleven Dorset rams were slaughtered after reaching puberty and total RNA was extracted from samples of semitendinosus and splenius muscles . P05019 mRNA was measured using a competitive reverse-transcription-polymerase chain reaction . P10275 and myostatin mRNA were measured by a ribonuclease protection assay ( RPA ) with standard curves . The means ( attomoles/microg RNA ) for splenius and semitendinosus muscles were 1.39 and 1.02 ( SE = 0.14 ) , 4.05 and 2.96 ( SE = 0.24 ) , and 4.30 and 3.85 ( SE = 0.37 ) for P05019 , AR , and myostatin , respectively . The difference between the two muscles was significant for P05019 and AR mRNA levels with higher levels in the splenius but not significant for myostatin . Our results show that locally produced P05019 and the regulation of AR expression may be important for sexually dimorphic muscle growth patterns . P10275 is responsible for rat organic cation transporter 2 gene regulation but not for rOCT1 and rOCT3 . PURPOSE : Organic cation transporters 1-3 ( OCT1-3 ; Slc22a1-3 ) mediate the membrane transport of organic cations in the kidney . We previously reported that rat (r)OCT2 expression in the kidney was regulated by testosterone . In this study , we examined the transcriptional mechanisms underlying the testosterone-dependent regulation of rOCT2 expression . METHODS : Approximately 3000-bp fragments of the rOCT1-3 promoter region were isolated , and promoter activities were measured in the renal epithelial cell line LLC- P30613 with the coexpression of rat androgen receptor . RESULTS : Among reporter constructs tested , only rOCT2 promoter activity was stimulated by testosterone . This stimulation was suppressed by nilutamide , an antiandrogen drug . Reporter assays using deletion constructs and mutational constructs of putative androgen response elements ( ARE ) in the rOCT2 promoter region suggested that two AREs , located at approximately -3000 and -1300 , respectively , play an important role in the induction by testosterone . CONCLUSIONS : DB00624 induces the expression of rOCT2 , but not of rOCT1 and rOCT3 , via the AR-mediated transcriptional pathway . This is the first study to address the transcriptional mechanisms of testosterone-dependent gene regulation of the Slc22 family . Sex steroid regulation of chin-marking behavior in male New Zealand rabbits . Chin-marking behavior ( chinning ) was evaluated daily in nine intact adult male rabbits . All subjects ( Ss ) displayed chinning ( mean of means +/- SE = 61 +/- 7 marks/10 min ) but the frequency of this behavior varied largely across them ( range of mean chinning frequency = 19-84 marks/10 min ) . Chinning frequency showed abrupt variations at intervals of 2-3 days , but periodogram analysis did not reveal the existence of an endogenous rhythm in this behavior . Castration significantly decreased ( mean of means +/- SE = 29 +/- 9 marks/10 min ; p < 0.01 ) . but did not suppress chinning . DB00624 propionate ( TP ; 1 mg/day for 16 days ) restored chinning in castrated Ss to slightly below precastration levels ( mean +/- S.E. V 53 +/- 13 marks/10 min ) . The daily administration of 1 microgram estradiol benzoate ( EB ) plus 1 mg dihydrotestosterone propionate ( DHTP ) stimulated chinning within 2 days ( mean increase = 147 % ; p < 0.005 ) . DHTP ( 1 mg/day ) given alone stimulated chinning only after 11 days of treatment ( mean increase = 475 % ; p < 0.01 ) . At higher doses , both DHTP ( 10 mg/day ) and EB ( 10 or 50 micrograms/day ) stimulated chinning by 450 % , 80 % , and 100 % , respectively , over baseline values . Results indicate that chinning largely depends on testicular steroids . P10275 occupation by T or DB02901 , which is enhanced by E , optimally activates chinning . P10275 in nuclei of rat testis . Testis nuclei of hypophysectomized rats selectively accumulate labeled testosterone and 5alpha-dihydrotestosterone following the injection of tritiated testosterone in vivo . DB00624 and 5alpha-dihydrotestosterone are bound to macromolecules in nuclei and can be extracted with 0.5 M DB00761 . Accumulation of protein bound radioactive androgens in nuclei of isolated seminiferous tubules is similar to that of whole testis . The relative amounts of testosterone and dihydrotestosterone in purified nuclei were similar to the relative amounts bound to cytoplasmic receptors , suggesting that cytoplasmic androgen-receptor complexes may be transported into the nuclei . Binding of labeled androgen is saturable and inhibited by prior injection of unlabeled testosterone or cyproterone acetate . Nuclear binding sites are destroyed by the proteolytic enzyme pronase , but not by DNase . Like the cytoplasmic androgen-receptor complexes in rat testis , nuclear androgen-protein complexes are heat labile and dissociate slowly at 0 degrees C. androgens fail to accumulate in testis nuclei of the Stanley-Gumbreck androgen insensitive rat , a species lacking cytoplasmic androgen receptors in testis and other androgen target tissues .

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P2y receptor-mediated angiogenesis via vascular endothelial growth factor receptor 2 signaling . Pathological as well as physiological angiogenesis is known to be regulated by such factors as nucleotides and Vascular Endothelial Growth Factor ( P15692 ) . Activated P2Y nucleotide receptors have been observed to associate and transactivate P15692 Receptor 2 ( P35968 ) , suggesting a cooperation between nucleotide and P15692 signaling in angiogenesis . P2YR mediated P35968 signaling therefore may be important in describing the angiogenic signaling of nucleotides such as DB00171 . Here , we provide evidence that supports the notion of P2YR- P35968 signaling . The significant angiogenic effect of P47900 /2 receptor agonists ( 100 microM DB00171 and 10 microM 2MS- DB00171 ) on endothelial cell tubulogenesis was suppressed back to near control levels upon addition of 1 microM SU1498 ( specific P35968 tyrosine kinase inhibitor ) . We believe that this P2YR-VEFGR2 signaling is an important component of pathological , as well as physiological angiogenesis . Shear stress-mediated F1/FO DB00171 synthase-dependent CO2 gas excretion from human pulmonary arteriolar endothelial cells . We studied the physiological role of flow through pulmonary arterioles in CO(2) gas exchange . We established human pulmonary arteriolar endothelial cells ( HPAoEC ) . The cells demonstrated marked immunocytochemical staining of P16284 , P15692 R2 , P12821 -1 , and CA type IV on their cell surface . Ten seconds shear stress stimulation caused the co-release of H(+) and DB00171 via the activation of F(1)/F(O) DB00171 synthase on the HPAoEC . F(1)/F(O) DB00171 synthase was immunocytochemically observed on the cell surface of non-permeabilized HPAoEC . In the shear stress-loaded HPAoEC culture media supernatant , ATPase activity increased in a time-dependent manner . The HPAoEC were strongly stained for P49961 , which partially co-localized with purinergic P47900 . The purinergic P47900 receptor agonist UTP ( 10(-6) M ) significantly potentiated the shear stress-induced increase in ATPase activity in the culture medium supernatant . Ten seconds shear stress stimulation also produced stress strength-dependent CO(2) gas excretion from the HPAoEC , which was significantly reduced by the inhibition of F(1)/F(O) DB00171 synthase or CA IV on the endothelial cell ( EC ) surface . In conclusion , we have proposed a new concept of CO(2) exchange in the human lung , flow-mediated F(1)/F(O) DB00171 synthase-dependent H(+) secretion , resulting in the facilitation of a dehydration reaction involving HCO3(-) in plasma and the excretion of CO(2) gas from arteriolar ECs . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Purinergic receptors involved in the immunomodulatory effects of DB00171 in human blood . We recently showed that the physiological compound DB00171 simultaneously inhibited P01375 and stimulated P22301 release in LPS-PHA stimulated blood . The purpose of the present study was to determine the mechanism involved in the concerted modulatory effect of DB00171 on P01375 and P22301 . Incubation of blood with DB00171 in the presence of selective P2 receptor antagonists showed that the stimulatory effect of DB00171 on P22301 release was completely annihilated by both 2-MeSAMP ( a Q9H244 /13 receptor antagonist ) and PSB-0413 ( a Q9H244 receptor antagonist ) . On the other hand , the inhibitory effect of DB00171 on P01375 release was completely reversed by 5'- P30566 ( a Q96G91 receptor antagonist ) as well as by H-89 , an inhibitor of DB02527 -activated PKA . The concerted inhibition by DB00171 of P01375 release via Q96G91 activation and stimulation of P22301 release via Q9H244 activation implicates a novel approach towards immunomodulation by altering the balance among pro- and anti-inflammatory cytokines . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Enhanced P47900 receptor expression in the brain after sensitisation with DB01576 . RATIONALE AND OBJECTIVES : Many pathological and physiological processes are associated with the transcriptional induction of specific receptors . The aim of the present study was to examine whether the development of DB01576 ( P49418 ) -induced sensitisation is related to an altered P2Y(1) receptor expression . METHODS : Rats , treated for 5 successive days with P49418 ( 1.5 mg/kg , i.p. ) , alone or after pre-treatment with the non-specific P2 receptor antagonist pyridoxal-phosphate-6-azophenyl-2,4-disulphonic acid ( PPADS , 0.6 nmol , i.c.v. ) and tested in an open field system with respect to locomotor response , were studied immunocytochemically 5 days after the last P49418 injection . RESULTS : In the behaviourally sensitised animals , astrogliosis , characterised by hypertrophy , increase in glial fibrillary acidic protein ( P14136 ) immunoreactivity ( IR ) and astrocytic proliferation in striatal areas and the nucleus accumbens were observed . Quantification of the P2Y(1) receptor stained cells revealed an increase in the receptor expression after P49418 -induced sensitisation in the studied regions . Pre-treatment with PPADS prior to each P49418 administration prevented the development of sensitisation , astrogliosis and P2Y(1) receptor up-regulation . PPADS failed to alter the number of P2Y(1) receptor-labelled cells when given alone . Confocal laser scanning microscopy indicated the localisation of P2Y(1) receptors on P14136 -labelled astrocytes as well as on tubulin ( betaIII ) -labelled neurones , under control conditions and after P49418 administration . CONCLUSION : The present results confirm the existence of P2Y(1) receptors on astrocytes and neurones as possible targets of endogenous DB00171 and in addition show their up-regulation as a consequence of P2Y(1) receptor-involvement in P49418 -induced sensitisation in vivo . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . DB00945 antagonizes the cytotoxic effect of methotrexate in lung cancer cells . DB00563 ( MTX ) has been widely used for the treatment of cancer and rheumatoid arthritis ( RA ) . DB00945 ( ASA ) is a non-selective cyclooxygenase ( P36551 ) inhibitor that contributes to the treatment of inflammatory conditions such as RA . It has been observed that the antitumor effect of ASA can be attributed to inhibition of cell cycle progression , induction of apoptosis and inhibition of angiogenesis . In the present study , we revealed that the treatment with a combination of MTX and ASA resulted in antagonism of the cytotoxic effect as demonstrated by P50991 and colony formation assays . ASA alleviated the MTX-mediated S phase accumulation and recovered the P55008 phase . MTX-mediated accumulation of the S phase marker cyclin A was also alleviated by ASA . Notably , FAS protein levels were upregulated by MTX in A549 cells . The antagonism of MTX efficacy caused by ASA was accompanied by altered expression of caspase-3 , Bcl-2 and FAS but not dihydrofolate reductase ( P00374 ) . This suggests that the alteration of caspase-3 , Bcl-2 and FAS was involved in the antagonism between ASA and MTX . Exogenously added folic acid reversed the MTX-mediated P00374 inhibition following either MTX or MTX + ASA treatments . Most importantly , we demonstrated for the first time that the commonly used non-steroidal anti-inflammatory drug for headache ASA and possibly other P23219 /2 inhibitors can produce a strong antagonistic effect on the growth inhibition of lung cancer cells when administered in combination with MTX . The clinical implication of our finding is obvious , i.e. , the clinical efficacy of MTX therapy can be compromised by ASA and their concomitant use should be avoided . ADP receptors -- targets for developing antithrombotic agents . Platelet P2 receptors -- P47900 , Q9H244 , and P51575 -- constitute the means by which adenine nucleotides can activate platelets . Coactivation of the Galphaq-coupled P47900 and Galphai2-coupled Q9H244 receptors is necessary for ADP-mediated platelet activation , which forms the basis of using P2 antagonists as antithrombotic drugs . P47900 receptor antagonists inhibit platelet activation , while P47900 knockout mice show longer bleeding times than normal mice but few other problems ; however , its ubiquitous expression in other tissues renders P47900 questionable as an antithrombotic target . The Q9H244 receptor is expressed nearly exclusively in platelets and brain , making it an attractive antithrombotic target . Antagonists for the Q9H244 receptor have been developed that either require metabolic activation to covalently inhibit Q9H244 and are irreversible , or simply are competitive in nature and thus reversible . DB00208 and clopidogrel are irreversible Q9H244 antagonists and have been repeatedly proven as clinical antithrombotic agents . In addition , a recently reported Q9H244 antagonist , CS-747 , shows promise as a future antithrombotic drug . The AR-C series of compounds represent reversible Q9H244 antagonists and have been used extensively to characterize the function of Q9H244 in platelets . Clinical studies show that AR-C69931MX is as effective as clopidogrel ; furthermore , the combination of AR-C69931MX ( cangrelor ) and clopidogrel confers greater antagonism of Q9H244 than either antagonist alone . The P51575 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change , and its inhibition or loss has little if any effect on hemostasis . A combination of P47900 and Q9H244 antagonists may represent an additional course of antithrombotic treatment . Effect of clopidogrel on circulating biomarkers of angiogenesis and endothelial activation . Angiogenic cytokines have been shown to influence vessel injury , and platelets represent a disposable circulating pool of angiogenic molecules . In the present study , objectives were to determine whether clopidogrel could have a potential effect on levels of circulating biomarkers of angiogenesis and endothelial activation . We explored 28 healthy white male volunteers treated for 7 days with clopidogrel 75 mg/day . We quantified angiogenic growth factors that have been shown to be correlated to cardiovascular risk or endothelial progenitor cell mobilization such as vascular endothelial growth factor ( P15692 ) -A and its soluble receptor forms P17948 and P35968 , placenta growth factor , and stromal cell-derived factor-1 . We also quantified soluble P16581 and P04275 to evaluate endothelial activation . Blood samples were drawn just before the first clopidogrel intake on day 1 , and after the last dosing ( day 7 ) . As expected , we observed a decrease in platelet reactivity in response to clopidogrel , confirmed by vasodilator-stimulated phosphoprotein phosphorylation assay . However , the 7-day intake of clopidogrel did not significantly modify the levels of the selected angiogenic factors or biomarkers of endothelial activation . These results show that circulating angiogenic factor level in healthy subjects is not driven by Q9H244 platelet receptor-induced activation and clopidogrel does not modify in a significant way the endothelial activation level . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . Effect of cyclooxygenase inhibitors on the P06850 -induced pituitary-adrenocortical activity during crowding stress . The aim of the present study was to determine the effect of social stress and significance of prostaglandins ( PG ) generated by constitutive and inducible cyclooxygenase ( P23219 and P35354 ) in the stimulation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis by corticotropin releasing hormone ( P06850 ) under basal and social crowding stress conditions . The stressed rats were crowded in groups of 24 to a cage for 3 or 7 days , whereas the control animals were haused in groups of 7 to a cage of the same size . The activity of Q9Y251 axis was determined by measuring plasma DB01285 and serum corticosterone levels 1 h after i.p . P06850 administration . Inhibitors of P23219 , piroxicam ( 0.2 , 2.0 , and 5.0 mg/kg ) , and P35354 , compound NS-398 ( 0.2 and 2.0 mg/kg ) , were administered i.p . 15 min prior to P06850 ( 0.1 microg/kg i.p. ) to control or crowded rats . The obtained results indicate that social stress for 3 and 7 days markedly intensifies the stimulatory action of P06850 on DB01285 secretion . Neither piroxicam nor NS-398 induce any significant effect on the P06850 -elicited DB01285 and corticosterone secretion in non-stressed or crowded rats . Therefore , PG generated by P23219 or P35354 do not participate to a significant extent in the stimulation of Q9Y251 axis by P06850 under either basal conditions or during crowding stress . These results also indicate that the stimulatory action of P06850 on DB01285 secretion is not only completely resistant to desensitization but is sensitized during social crowding stress . The results contrast with a significant involvement of PG in the vasopressin-induced stimulation of Q9Y251 response during crowding stress . Hypertonic stress regulates T cell function via pannexin-1 hemichannels and P2X receptors . Hypertonic saline ( HS ) resuscitation increases T cell function and inhibits posttraumatic T cell anergy , which can reduce immunosuppression and sepsis in trauma patients . We have previously shown that HS induces the release of cellular DB00171 and enhances T cell function . However , the mechanism by which HS induces DB00171 release and the subsequent regulation of T cell function by DB00171 remain poorly understood . In the present study , we show that inhibition of the gap junction hemichannel pannexin-1 ( Panx1 ) blocks DB00171 release in response to HS , and HS exposure triggers significant changes in the expression of all P2X-type DB00171 receptors in Jurkat T cells . Blocking or silencing of Panx1 or of P51575 , Q99571 , or Q99572 receptors blunts HS-induced p38 MAPK activation and the stimulatory effects of HS on TCR/ P10747 -induced P60568 gene transcription . Moreover , treatment with HS or agonists of P2X receptors overcomes T cell suppression induced by the anti-inflammatory cytokine P22301 . These findings indicate that Panx1 hemichannels facilitate DB00171 release in response to hypertonic stress and that P51575 , Q99571 , and Q99572 receptor activation enhances T cell function . We conclude that HS and P2 receptor agonists promote T cell function and thus , could be used to improve T cell function in trauma patients . Inhibition of P13671 rat glioma proliferation by [ Ru2Cl(Ibp)4 ] depends on changes in P38936 , p27 , Bax/Bcl2 ratio and mitochondrial membrane potential . The ruthenium compound [ Ru(2)Cl(Ibp)(4) ] ( or RuIbp ) has been reported to cause significantly greater inhibition of P13671 glioma cell proliferation than the parent HIbp . The present study determined the effects of 0-72h exposure to RuIbp upon P13671 cell cycle distribution , mitochondrial membrane potential , reactive species generation and mRNA and protein expression of Q01094 , cyclin D1 , c-myc , P06400 , P38936 , p27 , p53 , P12956 , P13010 , Bax , Bcl2 , cyclooxygenase 1 and 2 ( P23219 and P35354 ) . The most significant changes in mRNA and protein expression were seen for the cyclin-dependent kinase inhibitors P38936 and p27 which were both increased ( p < 0.05 ) . The marked decrease in mitochondrial membrane potential ( p < 0.01 ) and modest increase in apoptosis was accompanied by a decrease in anti-apoptotic Bcl2 expression and an increase in pro-apoptotic Bax expression ( p < 0.05 ) . Interestingly , P23219 expression was increased in response to a significant loss of prostaglandin E(2) production ( p < 0.001 ) , most likely due to the intracellular action of Ibp . Future studies will investigate the efficacy of this novel ruthenium-ibuprofen complex in human glioma cell lines in vitro and both rat and human glioma cells growing under orthotopic conditions in vivo . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice . Electrolyzed reduced water ( ERW ) is widely used for drinking by people in Asia . The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae . In the non-infected groups , interleukin ( IL ) -4 ( p < 0.001 ) , P05113 , P22301 , IL-1beta , tumor necrosis factor ( P01375 ) -alpha and immunoglobulin ( Ig ) A expression of the group fed ERW ( ERW group ) increased in small intestine compared with the normal control group . In the case of infected groups , the group fed ERW ( ERW+E. hortense group ) showed the result that P05112 , P05113 , P22301 and Ig A expression increased , but IL-1beta and P01375 ( p < 0.001 ) decreased , and the number of goblet cells ( p < 0.001 ) and helix pomatia agglutinin ( Q9Y251 ) positive cells increased compared with the group without feeding ERW . However , adult worm recovery rate was markedly increased ( p < 0.05 ) . On the other hand , the expression of all the cytokines except P22301 in spleen was mildly increased but not significant statistically , and there was no significant difference in the numerical changes of white blood cell ( WBC ) . These results indicate that feeding ERW may have influence on the local immune response ( Th-1 type cytokines such as IL-1beta , P01375 ) in the small intestine but not on the systemic immune response . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Dietary conjugated linoleic acid enhances spleen P37231 mRNA expression in broiler chicks . 1. The anti-inflammatory effects of dietary conjugated linoleic acid ( DB01211 ) on broilers repeatedly challenged with lipopolysaccharide ( LPS ) were investigated . 2 . Day-old broiler chicks were allotted into three treatment groups and fed on a control diet or diets containing 5.0 or 10.0 g DB01211 /kg diet . Six chicks from each treatment were injected with LPS ( 0.25 mg/kg body weight ) at 16 , 18 and 20 d of age . Splenic cyclooxygenase ( P36551 ) and inducible nitric oxide synthase ( P35228 ) activities , and prostaglandin E(2) ( PGE(2) ) and nitric oxide ( NO ) production as well as peroxisome proliferator-activated receptor-gamma ( P37231 ) mRNA expression were measured at 21 d of age . 3 . Chicks fed 10.0 g DB01211 /kg diet had lower P36551 activities and PGE(2) production that the controls . Dietary DB01211 ( 10.0 g/kg ) did not significantly diminish LPS-induced enhancement of COS-2 activity , inhibited the subsequent increase in PGE(2) production . 4 . Regulation of P23219 activity contributed to the difference in PGE(2) production . 5 . DB01211 did not markedly attenuate the increase of P35228 activity and NO production caused by LPS challenge . Chicks fed DB01211 had lower P35228 activity and NO production than the controls . 6 . Dietary DB01211 activated splenic P37231 mRNA expression and increased P37231 mRNA expression after LPS injection . 7 . These results suggest that dietary DB01211 has immunomodulatory effects in the spleen by restricting basal PGE(2) and NO to lower levels and enhancing P37231 mRNA expression . During the inflammatory response , dietary DB01211 did not alleviate the increase in P35354 and P35228 activities but enhanced Q06203 -gamma mRNA expression . Erosive arthritis and hepatic granuloma formation induced by peptidoglycan polysaccharide in rats is aggravated by prasugrel treatment . Administration of the thienopyridine Q9H244 receptor antagonist , clopidogrel , increased the erosive arthritis induced by peptidoglycan polysaccharide ( PG-PS ) in rats or by injection of the arthritogenic K/BxN serum in mice . To determine if the detrimental effects are caused exclusively by clopidogrel , we evaluated prasugrel , a third-generation thienopyridine pro-drug , that contrary to clopidogrel is mostly metabolized into its active metabolite in the intestine . Prasugrel effects were examined on the PG-PS-induced arthritis rat model . Erosive arthritis was induced in Lewis rats followed by treatment with prasugrel for 21 days . Prasugrel treated arthritic animals showed a significant increase in the inflammatory response , compared with untreated arthritic rats , in terms of augmented macroscopic joint diameter associated with significant signs of inflammation , histomorphometric measurements of the hind joints and elevated platelet number . Moreover , fibrosis at the pannus , assessed by immunofluorescence of connective tissue growth factor , was increased in arthritic rats treated with prasugrel . In addition to the arthritic manifestations , hepatomegaly , liver granulomas and giant cell formation were observed after PG-PS induction and even more after prasugrel exposure . Cytokine plasma levels of P01584 , P05231 , MIP1 alpha , MCP1 , Q16552 and RANTES were increased in arthritis-induced animals . P22301 plasma levels were significantly decreased in animals treated with prasugrel . Overall , prasugrel enhances inflammation in joints and liver of this animal model . Since prasugrel metabolites inhibit neutrophil function ex-vivo and the effects of both clopidogrel and prasugrel metabolites on platelets are identical , we conclude that the thienopyridines metabolites might exert non-platelet effects on other immune cells to aggravate inflammation . Gq-mediated Akt translocation to the membrane : a novel PIP3-independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol-3,4,5-trisphosphate ( PIP3 ) -dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5'-diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3-kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K/PIP3/Gi-independent mechanism . An Akt scaffolding protein , P38936 -activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi/PI3K/PIP3-independent mechanism . Glial cells , but not interstitial cells , express Q99572 , an ionotropic purinergic receptor , in rat gastrointestinal musculature . Purinergic ( DB00171 ) neurotransmission is a component of the inhibitory response of the musculature in various regions of the gastrointestinal tract . So far , seven ionotropic purinergic receptors ( P51575 -7 ) have been cloned . As specific antibodies become available , their respective distribution in the gastrointestinal tract can be elucidated . Here , we used high-resolution tricolor confocal microscopy , to study the distribution of Q99572 -immunoreactive ( -ir ) cells in the muscularis propria of the rat stomach , small intestine , and colon . Smooth muscle cells , P10721 -ir interstitial cells of Cajal , and P28906 / Q9UGI6 -ir fibroblastlike cells were Q99572 -negative , whereas Q99572 immunoreactivity was observed in nerves and S100-ir glial cells . In all regions studied , Q99572 immunoreactivity was also observed in myenteric and submucosal ganglia , where perineuronal nerve endings appeared brightly labeled . Our observations suggest that purinergic signaling could influence the enteric glia through Q99572 receptors . Expression of cyclooxygenase enzymes in rat hypothalamo-pituitary-adrenal axis : effects of endotoxin and glucocorticoids . Prostaglandins play a key role in mediating the hypothalamo-pituitary-adrenocortical ( Q9Y251 ) responses to immune insults . This study aimed to provide some insight into the relative contributions of the constitutive and inducible forms of cyclooxygenase ( P23219 and P35354 ) to the generation of these prostanoids by examining the effects of ( 1 ) endotoxin treatment on the expression of P23219 and P35354 mRNAs in the various components of the Q9Y251 axis in control and glucocorticoid pretreated rats , and ( 2 ) selective inhibition of P35354 on the production of corticosterone by adrenal tissue in vitro . Endotoxin caused a marked rise in P35354 mRNA in the adrenal gland that was evident 3 and 6 h after the injection and was prevented by pretreatment with dexamethasone . It also induced a modest increase in P35354 mRNA in the hypothalamus but not in the hippocampus or anterior pituitary gland . By contrast , P23219 mRNA was largely unaffected by the drug treatments in all tissues studied . In vitro the selective P35354 inhibitor SC-236 caused a marked reduction in adrenocorticotropic hormone-driven corticosterone release , as did the nonselective P36551 inhibitor , indomethacin . These results support a role of P35354 in the manifestation of the Q9Y251 responses to endotoxin , particularly within the adrenal gland . Identification of a variant in P35968 associated with serum P35968 and pharmacodynamics of DB06589 . PURPOSE : P15692 receptor ( VEGFR ) kinases are important drug targets in oncology that affect function of systemic endothelial cells . To discover genetic markers that affect VEGFR inhibitor pharmacodynamics , we performed a genome-wide association study of serum soluble vascular P35968 concentrations [ sVEGFR2 ] , a pharmacodynamic biomarker for P35968 inhibitors . EXPERIMENTAL DESIGN : We conducted a genome-wide association study ( GWAS ) of [ sVEGFR2 ] in 736 healthy Old Order Amish volunteers . Gene variants identified from the GWAS were genotyped serially in a cohort of 128 patients with advanced solid tumor with baseline [ sVEGFR2 ] measurements , and in 121 patients with renal carcinoma with [ sVEGFR2 ] measured before and during pazopanib therapy . RESULTS : rs34231037 ( C482R ) in P35968 , the gene encoding sVEGFR2 was found to be highly associated with [ sVEGFR2 ] , explaining 23 % of the variance ( P = 2.7 × 10(-37) ) . Association of rs34231037 with [ sVEGFR2 ] was replicated in 128 patients with cancer with comparable effect size ( P = 0.025 ) . Furthermore , rs34231037 was a significant predictor of changes in [ sVEGFR2 ] in response to pazopanib ( P = 0.01 ) . CONCLUSION : Our findings suggest that genome-wide analysis of phenotypes in healthy populations can expedite identification of candidate pharmacogenetic markers . Genotyping for germline variants in P35968 may have clinical utility in identifying patients with cancer with unusual sensitivity to effects of P35968 kinase inhibitors . A critical appraisal of the functional evolution of Q9H244 antagonists as antiplatelet drugs . Q9H244 receptor mediated inhibition of platelet aggregation is one of the most explored and exploited pathways in antiplatelet drug therapy to prevent ischemic events in patients undergoing percutaneous coronary intervention ( P05154 ) for the treatment of the acute coronary syndrome ( ACS ) . DB00208 , DB00758 , Prasugrel , DB08816 , DB06441 and Elinogrel are the Q9H244 inhibitors that act as antiplatelet drugs . In this review , the features of these drugs and the factors reported to be responsible for drug resistance or drug ineffectiveness were described . The features like drug metabolism , reversible or irreversible binding of drugs to their target protein and the mode of administration were observed to evolve along with the antiplatelet drugs . These features also include the drug-drug interactions , the pharmacogenetics and pharmacodynamics of Q9H244 inhibitors . We attempted to critically analyze how the desirable features were met by the Q9H244 inhibitors in the course of time . This review provides an overview of the evolution of Q9H244 inhibitors and may guide the researchers to develop better antiplatelet drugs in the future . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Characterization of mechanisms involved in secretion of active heparanase . Q9Y251 is an endo-beta-D-glucuronidase involved in extracellular matrix remodeling and degradation and implicated in tumor metastasis , angiogenesis , inflammation , and autoimmunity . The enzyme is synthesized as a latent 65-kDa protein and is processed in the lysosomal compartment to an active 58-kDa heterodimer , where it is stored in a stable form . In contrast , its heparan sulfate substrate is localized extracellularly , suggesting the existence of mechanisms that trigger heparanase secretion . Here we show that secretion of the active enzyme is mediated by the protein kinase A and C pathways . Moreover , secretion of active heparanase was observed upon cell stimulation with physiological concentrations of adenosine , ADP , and DB00171 , as well as by the noncleavable DB00171 analogue adenosine 5'-O-(thiotriphosphate) . Indeed , heparanase secretion was noted upon cell stimulation with a specific P47900 receptor agonist and was inhibited by P2Y receptor antagonists . The kinetics of heparanase secretion resembled the secretion of cathepsin D , a lysosomal enzyme , indicating that the secreted heparanase is of lysosomal origin . We suggest that secretion of active heparanase is initiated by extracellular cues activating the protein kinase A and C signaling pathways . The secreted enzyme(s) then facilitate cell invasion associated with cancer metastasis , angiogenesis , and inflammation . n-3 fatty acids specifically modulate catabolic factors involved in articular cartilage degradation . This study describes specific molecular mechanisms by which supplementation with n-3 fatty acids ( i.e. those present in fish oils ) can modulate the expression and activity of degradative and inflammatory factors that cause cartilage destruction during arthritis . Our data show that incorporation of n-3 fatty acids ( but not other polyunsaturated or saturated fatty acids ) into articular cartilage chondrocyte membranes results in a dose-dependent reduction in : ( i ) the expression and activity of proteoglycan degrading enzymes ( aggrecanases ) and ( ii ) the expression of inflammation-inducible cytokines ( interleukin ( IL ) -1alpha and tumor necrosis factor ( P01375 ) -alpha ) and cyclooxygenase ( P35354 ) , but not the constitutively expressed cyclooxygenase P23219 . These findings provide evidence that n-3 fatty acid supplementation can specifically affect regulatory mechanisms involved in chondrocyte gene transcription and thus further advocate a beneficial role for dietary fish oil supplementation in alleviation of several of the physiological parameters that cause and propogate arthritic disease . Proteomic profiling of cancer stem cells derived from primary tumors of P04626 /Neu transgenic mice . Human epidermal growth factor receptor 2 ( P04626 ) overexpression leads to mammary tumorigenesis and its elevated levels lead to increase in cancer stem cells ( CSCs ) , invasion , and metastasis . CSCs are resistant to radiation/chemotherapeutic drugs and are believed to be responsible for recurrence/relapse of cancer . CSCs are isolated using flow cytometry based sorting , although reliable , this technology hinders the convenient identification of molecular targets of CSCs . Therefore to understand the molecular players of increased CSC through P04626 overexpression and to develop meaningful targets for combination therapy , we isolated and characterized breast CSCs through convenient tumorsphere culture . We identified the altered protein expression in CSC as compared to non-CSC using LC-MS/MS and confirmed those results using qRT-PCR and Western blotting . P02794 1 ( P02794 ) was identified as a candidate gene , which is involved in iron metabolism and iron depletion significantly decreased the self-renewal of CSCs . We further performed in silico analysis of altered genes in tumorsphere and identified a set of genes ( P06454 , P26447 , P06703 , TNXRD1 , P23219 , P35354 , P02533 , and P02794 ) , representing possible molecular targets , which in combination showed a promise to be used as prognostic markers for breast cancer . Initial testing of the multitargeted kinase inhibitor pazopanib by the Pediatric Preclinical Testing Program . DB06589 is an oral angiogenesis inhibitor targeting vascular growth factor receptor-1 , -2 , and -3 , platelet derived growth factor receptor-α , platelet derived growth factor receptor-β , and P10721 that has demonstrated activity against a variety of adult cancer xenografts . DB06589 was tested against a panel of pediatric rhabdomyosarcoma and Ewing sarcoma xenografts at a dose of 108 mg/kg/day or 100 mg/kg twice daily , administered orally for 28 days . While no objective responses were observed , pazopanib induced statistically significant differences in event-free survival compared to controls in approximately one-half of the sarcoma xenograft models tested . Though well tolerated , pazopanib showed limited activity against the sarcoma models evaluated , with the best tumor responses being growth delay . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Platelet Q9H244 receptor inhibition by thienopyridines : status and future . Thienopyridines have a well-established role in the treatment of coronary artery disease , especially in the setting of acute coronary syndromes and percutaneous coronary interventions . DB00208 , the first FDA-approved thienopyridine , was shown to be effective in reducing coronary events in high risk patients , but the original enthusiasm was hampered by concerns about its serious bone marrow toxicity . DB00758 a second generation thienopyridine with lesser side effects , is not only at least as effective as ticlopidine , but in combination with a low dose of aspirin , has been demonstrated to reduce the risk of major cardiovascular events in acute coronary syndrome patients in large-scale , randomised trials . Recent studies have highlighted major flaws in clopidogrel pharmacokinetics due to its delayed onset of action , and much attention has been devoted to the phenomenon of clopidogrel ' resistance ' . Among the novel , third generation thienopyridines , prasugrel as compared to clopidogrel has demonstrated lower inter-patient response variability and a reduced incidence of ischaemic events , but at an increased risk of major bleeding . Currently , several studies are continuing to test new direct Q9H244 receptor antagonists , such as cangrelor and AZD6140 , characterised by a faster reversal of platelet inhibition . DB00184 consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 - P30532 - P30926 cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5(-/-) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5*-nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . [ Thienopyridines in the treatment and prevention of cardiovascular diseases. Part I. DB00208 ] . In a series of articles the authors consider clinical pharmacology and experience of clinical application of blockers of platelet Q9H244 receptors , most well known representatives of which ticlopidine and clopidogrel according to chemical structure belong to thienopyridine derivatives . In the first communication pharmacodynamics and pharmacokinetics of the first thienopyridine ticlopidine are described in detail . Results of randomized studies in which cerebro and cardioprotective efficacy and safety of ticlopidine was studied in patients with cerebrovascular , peripheral artery diseases , and acute coronary syndromes are discussed . It has been established that ticlopidine is more effective and safe in patients having undergone coronary and femoral bypass surgery . Results of meta analyses have shown which evidence that ticlopidine is not less and may be more effective than clopidogrel in patients after coronary bypass surgery . Most frequent and most severe side effects of ticlopidine and measures of their prevention are also considered . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Comparative effects of the anti-platelet drugs , clopidogrel , ticlopidine , and cilostazol on aspirin-induced gastric bleeding and damage in rats . AIMS : The present study compared the effects of frequently used anti-platelet drugs , such as clopidogrel , ticlopidine , and cilostazol , on the gastric bleeding and ulcerogenic responses induced by intraluminal perfusion with 25 mM aspirin acidified with 25 mM HCl ( acidified ASA ) in rats . MAIN METHODS : The stomach was perfused with acidified ASA at a rate of 0.4 ml/min for 60 min under urethane anesthesia , and gastric bleeding was measured as the concentration of hemoglobin in the luminal perfusate , which was collected every 15 min . DB00758 ( 10-100mg/kg ) , ticlopidine ( 10-300 mg/kg ) , or cilostazol ( 3-30 mg/kg ) was given p.o . 24h or 90 min before the perfusion of acidified ASA , respectively . KEY FINDINGS : Perfusion of the stomach with acidified ASA alone led to slight bleeding and lesions in the stomach . The pretreatment with clopidogrel , even though it did not cause bleeding or damage by itself , dose-dependently increased the gastric bleeding and ulcerogenic responses induced by acidified ASA . DB00208 also aggravated the severity of damage by increasing gastric bleeding , and the effects of ticlopidine at 300 mg/kg were equivalent to those of clopidogrel at 100mg/kg . In contrast , cilostazol dose-dependently decreased gastric bleeding and damage in response to acidified ASA . SIGNIFICANCE : These results demonstrated that clopidogrel and ticlopidine , Q9H244 receptor inhibitors , increased gastric bleeding and ulcerogenic responses to acidified ASA , to the same extent , while cilostazol , a phosphodiesterase III inhibitor , suppressed these responses . Therefore , cilostazol may be safely used in dual anti-platelet therapy combined with ASA , without increasing the risk of gastric bleeding . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . DB00563 induces apoptosis through p53/ P38936 -dependent pathway and increases P12830 expression through downregulation of HDAC/ Q15910 . DB00563 ( MTX ) is a dihydrofolate reductase ( P00374 ) inhibitor widely used as an anticancer drug in different kinds of human cancers . Here we investigated the anti-tumor mechanism of MTX against non-small cell lung cancer ( NSCLC ) A549 cells . MTX not only inhibited in vitro cell growth via induction of apoptosis , but also inhibited tumor formation in animal xenograft model . RNase protection assay ( RPA ) and RT-PCR demonstrated its induction of p53 target genes including DR5 , P38936 , Puma and Noxa . Moreover , MTX promoted p53 phosphorylation at Ser15 and acetylaion at Lys373/382 , which increase its stability and expression . The apoptosis and inhibition of cell viability induced by MTX were dependent on p53 and , partially , on P38936 . In addition , MTX also increased P12830 expression through inhibition of histone deacetylase ( HDAC ) activity and downregulation of polycomb group protein enhancer of zeste homologue 2 ( Q15910 ) . Therefore , the anticancer mechanism of MTX acts through initiation of p53-dependent apoptosis and restoration of P12830 expression by downregulation of HDAC/ Q15910 . Purinergic receptor ligands stimulate pro-opiomelanocortin gene expression in AtT-20 pituitary corticotroph cells . Although recent studies have suggested that purinergic receptors are expressed in the anterior pituitary gland , their involvement in the regulation of pituitary hormone gene expression is not completely understood . In the present study , we examined the expression of purinergic receptors and the effects of purinergic receptor ligands on pro-opiomelanocortin ( P01189 ) gene expression , in AtT20 mouse corticotroph cells . We identified the expression of most of the purinergic receptor subtypes ( A1 , A2 , P51575 , 3-7 , P47900 , 2 , 4 ) mRNAs , analysed by the reverse transcriptase-polymerase chain reaction . We also found that adenosine and DB00171 , two representative and endogenous agonists of A1-3 and P2X/P2Y receptors , respectively , stimulated the 5'-promoter activity of the P01189 gene in a dose- and time-related manner . When these ligands were simultaneously used with corticotrophin-releasing hormone ( P06850 ) , effects that were more than additive were observed , suggesting an enhancing role of these compounds in P06850 -mediated adrenocorticotrophic hormone ( DB01285 ) synthesis . These ligands also stimulated the expression of transcription factors involved in the regulation of the P01189 gene , but did not enhance DB01285 secretion . Finally , the positive effect of adenosine as well as P06850 was completely inhibited by the protein kinase A inhibitor H89 , whereas that of DB00171 was not influenced , indicating that different intracellular signalling pathways mediate these effects . Altogether , our results suggest a stimulatory role for these purinergic receptor ligands in the regulation of P01189 gene expression in corticotroph cells . Because adenosine and DB00171 are known to be produced within the pituitary gland , it is possible they may be acting in an autocrine/paracrine fashion . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . Purinergic receptors are part of a functional signaling system for proliferation and differentiation of human epidermal keratinocytes . We investigated the expression of Q93086 , Q99572 , P47900 and P41231 receptor subtypes in normal human epidermis and in relation to markers of proliferation ( P12004 and Ki-67 ) , keratinocyte differentiation ( cytokeratin P13645 and involucrin ) and markers of apoptosis ( TUNEL and anticaspase-3 ) . Using immunohistochemistry , we showed that each of the four receptors was expressed in a spatially distinct zone of the epidermis , suggesting different functional roles for these receptors . Functional studies were performed on primary cultures of human keratinocytes and on explanted rat skin , where different P2 receptor subtype agonists and antagonists were applied to cultured keratinocytes or injected subcutaneously into the skin , respectively . An increase in cell number was caused by low doses of the nonspecific P2 receptor agonist DB00171 , the P41231 receptor agonist UTP ( p < 0.001 ) , and the P47900 receptor agonist 2MeSADP ( p < 0.05 ) . There was a significant decrease in cell number as a result of treatment with the Q93086 receptor agonist ATPgammaS ( p < 0.001 ) and the Q99572 receptor agonist BzATP ( p < 0.001 ) . Suramin caused a significant block in the effect of 100 microm DB00171 ( p < 0.01 ) and 1000 microm DB00171 ( p < 0.001 ) on cell number . These results imply that different purinergic receptors have different functional roles in the human epidermis with P47900 and P41231 receptors controlling proliferation , while Q93086 and Q99572 receptors control early differentiation , terminal differentiation and death of keratinocytes , respectively . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . The thienopyridine derivatives ( platelet adenosine diphosphate receptor antagonists ) , pharmacology and clinical developments . The thienopyridines , ticlopidine and clopidogrel , are antiplatelet drugs . They are prodrugs and are metabolised in the liver to active metabolites that are non-competitive antagonists of the platelet adenosine diphosphate receptor , Q9H244 . Inhibition of platelet aggregation by these drugs is delayed until 24-48 h after administration , with maximal inhibition achieved after 3-5 days . Recovery of platelet function after drug withdrawal is slow ( 7-14 days ) . DB00208 and clopidogrel are effective in preventing atherothrombotic events in cardiovascular , cerebrovascular and peripheral vascular disease . Gastrointestinal side effects and skin rashes are common . However , neutropenia and thrombotic thrombocytopenic purpura are significant and sometimes fatal adverse effects of ticlopidine . DB00758 appears to offer several advantages over ticlopidine : a more rapid onset of action and a lower incidence of neutropenia and thrombotic thrombocytopenic purpura.A combination of clopidogrel and aspirin has become standard for antithrombotic therapy in cardiovascular disease . The anaesthetic considerations of patients taking the thienopyridine compounds are discussed . Purine receptor Q15077 mediates cellular response to γ-ray-induced DNA damage . We previously showed that nucleotide P2 receptor agonists such as DB00171 and UTP amplify γ-ray-induced focus formation of phosphorylated histone H2A variant P16104 ( γ P16104 ) , which is considered to be an indicator of DNA damage so far , by activating purine Q15077 and Q9H244 receptors . Therefore , we hypothesized that these P2 receptors play a role in inducing the repair response to γ-ray-induced DNA damage . In the present study , we tested this idea by using human lung cancer A549 cells . First , reverse-transcription polymerase chain reaction ( RT-PCR ) showed that Q15077 receptor is highly expressed in A549 cells , but Q9H244 receptor is only weakly expressed . Next , colony formation assay revealed that Q15077 receptor antagonist MRS2578 markedly reduced the survival rate of γ-ray-exposed A549 cells . The survival rate was also significantly reduced in Q15077 -knock-down cells , compared with scramble siRNA-transfected cells . Since it has reported that phosphorylation of P27361 /2 after activation of P00533 via Q15077 and Q9H244 receptors is involved in the repair response to γ-ray-induced DNA damage , we next examined whether γ-ray-induced phosphorylation of P27361 /2 was also inhibited by MRS2578 in A549 cells . We found that it was . Taken together , these findings indicate that purinergic signaling through Q15077 receptor , followed by P27361 /2 activation , promotes the cellular repair response to γ-ray-induced DNA damage .

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Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . P05231 -receptor polymorphisms rs12083537 , rs2228145 , and rs4329505 as predictors of response to tocilizumab in rheumatoid arthritis . DB06273 ( TCZ ) , a monoclonal antibody targeting the human interleukin-6-receptor ( IL-6R ) , is indicated for the treatment of rheumatoid arthritis ( RA ) . We examined whether three P08887 single-nucleotide polymorphisms rs12083537 , rs2228145 ( formerly rs8192284 ) , and rs4329505 with previously reported functional effects were associated with clinical response to TCZ in a retrospective study cohort consisting of 79 RA patients . Three months after initiation of TCZ therapy , changes in swollen joint count ( SJC ) and , subordinately , tender joint count ( TJC ) , serum-CRP , DAS28-CRP , and EULAR-response were tested for association with the P08887 -haplotype or genotype . The major allele ( A ) of rs12083537 and the minor allele ( C ) of rs4329505 were associated with a poor SJC response ( P=0.02 and 0.02 , respectively ) . Moreover , the AAC-haplotype ( for rs12083537 , rs2228145 , and rs4329505 , respectively ) was associated with a poor SJC response ( P=0.00004 ) and , with borderline significance , EULAR-response ( P=0.05 ) . These data suggest that genetic variation in P08887 may aid in predicting TCZ therapy outcome in RA patients . Levels of NT-proBNP , markers of low-grade inflammation , and endothelial dysfunction during spironolactone treatment in patients with diabetic kidney disease . P00797 -angiotensin-aldosterone system ( RAAS ) blockade may reduce levels of biomarkers of chronic low-grade inflammation and endothelial dysfunction . We investigated the effect of spironolactone added to standard RAAS blockade on these biomarkers in an analysis of four original studies . MATERIALS AND METHODS : The studies were double-blind , randomised , placebo-controlled studies in 46 type 1 and 23 type 2 diabetic patients with micro- or macroalbuminuria treated with angiotensin-converting enzyme inhibitor ( P12821 inhibitor ) or angiotensin receptor blocker ( ARB ) , and randomised to additional treatment with spironolactone 25 mg and placebo daily for 60 days . OUTCOME MEASURES : Changes in inflammatory ( hsCRP , s-ICAM , P01375 α , P05231 , P10145 , Serum amyloid A , IL1β ) , endothelial dysfunction ( sE-selectin , s- P05362 , s- P19320 , P04275 , p-selectin , s-thrombomodulin ) and NT-proBNP after each treatment period . RESULTS : During spironolactone treatment , u-albumin excretion rate was reduced from 605 ( 411-890 ) to 433 ( 295-636 ) mg/24 h , as previously reported . Markers of inflammation and endothelial dysfunction did not change ; only changes in NT-proBNP ( reduced by 14 % , p=0.05 ) and serum amyloid A ( reduced by 62 % , p=0.10 ) were borderline significant . DISCUSSIONS : Our results indicate that the renoprotective effect of spironolactone when added to RAAS blockade is not mediated through anti-inflammatory pathways since markers of inflammation and endothelial dysfunction are not affected during treatment . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Gene transfer of GM- P04141 , P33681 and CD154 cDNA enhances survival in a murine model of acute leukemia with persistence of a minimal residual disease . Gene transfer of various cytokines and co-stimulatory molecules has been reported to induce a potent antileukemic immunity in murine models , however , the relative efficiency and possible synergistic effects between candidate genes have not been extensively investigated . We analyzed in a murine model of P11274 / P00519 acute leukemia whether gene transfer of CD154 , P33681 or GM- P04141 as a single agent or combination of CD154 + GM- P04141 , P33681 + CD154 and GM- P04141 + P33681 in leukemic cells could enhance survival . We observed that CD154 gene transfer induced a marked inhibition of leukemogenicity , and also that CD154 and combination of GM- P04141 and P33681 gene transfer protected mice against subsequent challenge with leukemic cells and had a therapeutic effect for a pre-established leukemia disease . We also found minimal residual leukemic disease by RT-PCR for 6 to 12 months in 0 to 25 % of animals injected with transduced leukemic cells and surviving the challenge without evidence of disease , except in the control empty plasmid group where very few mice survived the challenge but all of those were positive by RT-PCR . These findings suggest that leukemic cell vaccination by gene transfer can induce a tumor dormancy phenomenon compatible with long-term survival . Neisseria meningitidis capsular polysaccharides induce inflammatory responses via O60603 and O00206 -MD-2 . CPS are major virulence factors in infections caused by Neisseria meningitidis and form the basis for meningococcal serogroup designation and protective meningococcal vaccines . CPS polymers are anchored in the meningococcal outer membrane through a 1,2-diacylglycerol moiety , but the innate immunostimulatory activity of CPS is largely unexplored . Well-established human and murine macrophage cell lines and P29320 /TLR stably transfected cells were stimulated with CPS , purified from an endotoxin-deficient meningococcal serogroup B NMB-lpxA mutant . CPS induced inflammatory responses via O60603 - and O00206 -MD-2 . Meningococcal CPS induced a dose-dependent release of cytokines ( P01375 -α , P05231 , P10145 , and P02778 ) and NO from human and murine macrophages , respectively . CPS induced P10145 release from P29320 cells stably transfected with O60603 /6 , O60603 , O60603 / P08571 , and O00206 /MD-2/ P08571 but not P29320 cells alone . mAb to O60603 but not an isotype control antibody blocked CPS-induced P10145 release from P29320 - O60603 /6-transfected cells . A significant reduction in P01375 -α and P10145 release was seen when THP-1- and P29320 - O00206 /MD-2- P08571 - but not P29320 - O60603 - or P29320 - O60603 /6-transfected cells were stimulated with CPS in the presence of DB04933 ( E5564 ) , a lipid A antagonist that binds to MD-2 , and a similar reduction in NO and P01375 -α release was also seen in RAW 264.7 cells in the presence of DB04933 . P08571 and P18428 enhanced CPS bioactivity , and NF-κB was , as anticipated , the major signaling pathway . Thus , these data suggest that innate immune recognition of meningococcal CPS by macrophages can occur via O60603 - and O00206 -MD-2 pathways . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . DB06273 in pediatric rheumatology : the clinical experience . During the last two decades , clinical use of novel biological therapy has led to increased mechanistic understanding of complex rheumatological diseases . Conversely , basic and translational studies have led to development of new and varied therapeutic agents . These new medications which " target " specific steps in one or more immune pathways have the potential to control disease symptoms , improve quality of life and long-term prognosis , and perhaps in some , restore immunological tolerance . Use of these agents in clinical trials , combined with post-marketing surveillance , has revealed both the benefits and the undesirable side-effects of biological disease-modifying anti-rheumatic drugs ( DMARDs ) . In this review we focus on the use of tocilizumab , a monoclonal antibody directed against the P05231 receptor ( P08887 ) , which potently inhibits P05231 / P08887 signaling . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Cerebrospinal fluid synaptic proteins as useful biomarkers in tyrosine hydroxylase deficiency . Tyrosine hydroxylase ( TH ) deficiency is an inborn error of dopamine biosynthesis and a cause of early parkinsonism . Two clinical phenotypes have been described . Type " B " : early onset severe encephalopathy ; type " A " : later onset , less severe and better response to L-dopa . We aimed to study the expression of several key dopaminergic and gabaergic synaptic proteins in the cerebrospinal fluid ( P04141 ) of a series of patients with TH deficiency and their possible relation with the clinical phenotype and response to DB01235 . Dopamine transporter ( Q01959 ) , D2-receptor and vesicular monoamine transporter ( Q05940 ) were measured in the P04141 of 10 subjects with TH deficiency by Western blot analysis . In 3 patients , data of pre- and post-treatment with DB01235 were available , and in one of them , GABA vesicular transporter was determined . Results were compared to an age-matched control population . The concentration of D2-receptors in P04141 was significantly higher in patients with TH deficiency than in controls . Similarly , Q01959 and vesicular monoamine transporter type 2 were up-regulated . Studies performed before DB01235 , and on DB01235 therapy showed a paradoxical response with D2 receptor expression increase as L-Dopa doses and homovanillic concentration gradually raised in a B phenotype patient . The opposite results were found in two patients with A phenotype . However , this is a very small sample , and further studies are needed to conclude robust differences between phenotypes . Synaptic proteins are detectable in the P04141 and their quantification can be useful for understanding the pathophysiology of neurotransmitter defects and potentially to adjust and personalize treatments in the future . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . P12821 ( P12821 ) and Q9BYF1 levels in the cerebrospinal fluid of patients with multiple sclerosis . BACKGROUND : We reported a reduction in the levels of angiotensin II in cerebrospinal fluid ( P04141 ) from patients with multiple sclerosis ( MS ) . OBJECTIVE AND METHODS : To clarify the mechanism underlying this reduction , we assayed angiotensin-converting enzyme ( P12821 ) and Q9BYF1 concentrations along with angiotensin II concentrations in P04141 samples from 20 patients with MS and 17 controls with non-neurological diseases . RESULTS : P12821 levels were significantly elevated in patients with MS compared with controls ( 48.42 +/- 4.84 vs 44.71 +/- 3.9 pg/mL ) , whereas Q9BYF1 levels were significantly reduced ( 2.56 +/- 0.26 vs 2.78 +/- 0.24 pg/mL ) , acting toward a normalization of angiotensin II levels . CONCLUSION : These results further indicate an alteration of the intrathecal renin-angiotensin system in patients with MS . P37840 expression modulates microglial activation phenotype . Recent Parkinson 's disease research has focused on understanding the function of the cytosolic protein , alpha-synuclein , and its contribution to disease mechanisms . Within neurons , alpha-synuclein is hypothesized to have a role in regulating synaptic plasticity , vesicle release , and trafficking . In contrast , glial-expressed alpha-synuclein remains poorly described . Here , we examine the consequence of a loss of alpha-synuclein expression on microglial activation . Using a postnatal brain-derived culture system , we defined the phenotype of microglia from wild-type and knock-out alpha-synuclein mice ( Scna-/- ) . Scna-/- microglia displayed a basally increased reactive phenotype compared with the wild-type cells and an exacerbated reactive phenotype after stimulation . They also exhibited dramatic morphologic differences compared with wild-type , presenting as large , ramified cells filled with vacuole-like structures . This corresponded with increased protein levels of activation markers , P34810 and beta1 integrin , in the Scna-/- cells . More importantly , Scna-/- microglia , after stimulation , secreted elevated levels of proinflammatory cytokines , TNFalpha ( tumor necrosis factor alpha ) and P05231 ( interleukin-6 ) , compared with wild type . However , despite the reactive phenotype , Scna-/- cells had impaired phagocytic ability . We demonstrate for the first time that alpha-synuclein plays a critical role in modulating microglial activation state . We suggest that altered microglial alpha-synuclein expression will affect their phenotype as has already been demonstrated in neurons . This has direct ramifications for the contribution of microglia to the pathophysiology of disease , particularly in familial cases linked to altered alpha-synuclein expression . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . Expression of the adaptor protein Lnk in leukemia cells . OBJECTIVE : Tyrosine kinases are involved in cytokine signaling and are frequently aberrantly activated in hematological malignancies . Lnk , a negative regulator of cytokine signaling , plays critical nonredundant roles in hematopoiesis . By binding to phosphorylated tyrosine kinases , Lnk inhibits major cytokine receptor signaling , including c- P10721 ; erythropoietin receptor- O60674 ( O60674 ) ; and P40238 - O60674 . In the present study , we investigated Lnk expression and possible function in transformed hematopoietic cells . MATERIALS AND METHODS : Coimmunoprecipitations were performed to identify binding between Lnk and mutant tyrosine kinases . Proliferation assays were done to examine the affect of Lnk overexpression on cancer cell growth . Real-time polymerase chain reaction analysis was used to determine Lnk expression in patient samples . RESULTS : We show that , in parallel to binding wild-type O60674 and c- P10721 , Lnk associates with and is phosphorylated by mutant alleles of O60674 and c- P10721 . In contrast , Lnk does not bind to and is not phosphorylated by P11274 - P00519 fusion protein . Ectopic expression of Lnk strongly attenuates growth of some leukemia cell lines , while others as well as most solid tumor cancer cell lines are either moderately inhibited or completely insensitive to Lnk . Furthermore , Lnk-mediated growth inhibition is associated with differential downregulation of phosphatidylinositol 3 kinase/Akt/mammalian target of rapamycin and mitogen-activated protein kinase/extracellular signal-regulated kinase signaling in leukemia cell lines . Surprisingly , analysis of Lnk in a large panel of myelodysplastic syndrome and acute myeloid leukemia patient samples revealed high levels of Lnk in nearly half of the samples . CONCLUSION : Although how leukemic cells overcome the antiproliferative effects of Lnk is not yet clear , our data highlight the multifaceted role negative feedback mechanisms play in malignant transformation . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Anti- P05231 -receptor-alpha ( tocilizumab ) does not inhibit human monocyte-derived dendritic cell maturation or alloreactive T-cell responses . Significant comorbidites and lethality complicate GVHD and its treatment . Targeting the cytokine milieu may improve GVHD control ; and P05231 is an attractive candidate , given its role in dendritic cell activation and T-cell differentiation . DB06273 is a humanized mAb to P05231 -receptor-α ( P08887 -α ) , which is Food and Drug Administration-approved for treatment of rheumatoid arthritis . Mouse transplant models have demonstrated that P05231 blockade also improves GVHD scores and survival . Definitive immunologic effects of P05231 inhibition have not emerged given inconsistent alterations in regulatory T cells ( Tregs ) and suppression of T-cell proliferation . Despite on-target suppression of P08887 -α signaling in human monocyte-derived dendritic cells ( moDCs ) and T cells , our data show no effect on moDC maturation/activation , alloreactive T-cell proliferation , Treg expansion , or allogeneic Th1/Th17 responses in vitro . These findings merit attention in any clinical trials of tocilizumab for GVHD prevention or treatment and provide a rationale for evaluating more specific inhibitors of downstream O60674 / P40763 signaling as well .

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interacts_with DB00030?

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P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Adaptive responses to dasatinib-treated lung squamous cell cancer cells harboring Q16832 mutations . Q16832 mutations occur in approximately 4 % of lung squamous cell cancer ( SCC ) where the tyrosine kinase inhibitor dasatinib has emerged as a new therapeutic option . We found that P29323 and AKT phosphorylation was weakly inhibited by dasatinib in Q16832 -mutant lung SCC cells , suggesting that dasatinib inhibits survival signals distinct from other oncogenic receptor tyrosine kinases ( RTK ) and/or compensatory signals exist that dampen dasatinib activity . To gain better insight into dasatinib 's action in these cells , we assessed altered global tyrosine phosphorylation ( pY ) after dasatinib exposure using a mass spectrometry-based quantitative phosphoproteomics approach . Overlaying protein-protein interaction relationships upon this dasatinib-regulated pY network revealed decreased phosphorylation of Src family kinases and their targets . Conversely , dasatinib enhanced tyrosine phosphorylation in a panel of RTK and their signaling adaptor complexes , including P00533 , MET/ Q13480 , and P08069 / Q9Y4H2 , implicating a RTK-driven adaptive response associated with dasatinib . To address the significance of this observation , these results were further integrated with results from a small-molecule chemical library screen . We found that dasatinib combined with MET and insulin-like growth factor receptor ( P08069 ) inhibitors had a synergistic effect , and ligand stimulation of P00533 and MET rescued Q16832 -mutant lung SCC cells from dasatinib-induced loss of cell viability . Importantly , we observed high levels of tyrosine-phosphorylated P00533 and MET in a panel of human lung SCC tissues harboring Q16832 mutations . Our results highlight potential RTK-driven adaptive-resistant mechanisms upon Q16832 targeting , and they suggest new , rationale cotargeting strategies for Q16832 -mutant lung SCC . [ Treatment of type 1 diabetes mellitus revealed below 7 years of age in the Diabetes Center of Silesia , Poland ] . INTRODUCTION : Frequency of type 1 diabetes mellitus diagnosis in young children increases . Within this group , such factors as limited cooperation , little acceptance of multiple injections and other typical patterns of behavior can strongly influence the insulin management outcome . AIM OF THE STUDY : The objective of the study was to provide information regarding metabolic control in young diabetes patients . MATERIAL AND METHODS : Charts of 58 children with T1DM , all subjects under control of our Department , that were aged at onset ( 1998-2003 ) below 7 years ( mean 4.05+/-1.6 ) were studied retrospectively . HbA1c , total , bolus and basal daily insulin requirement ( P30518 ) , weight , height , severe hypoglycaemia and diabetic ketoacidosis ( DKA ) were analyzed till April 2006 in 2-year intervals . P01308 therapy model was also taken into consideration . RESULTS : Mean HbA1c was 7.2+/-1.2 % for all children for the whole studied period and did not alter significantly between analyzed intervals . Most common treatment model at diabetes onset was the therapy with premixed insulin ( Mix ) ( 67 % ) and after 4 and 6 years - continuous subcutaneous insulin infusion ( CSII ) ( 50 % and 75 % respectively ) . A tendency for a better metabolic control was observed at multiple daily injections and CSII than at Mix . Change of the weight or height percentile channel was not revealed . Bolus and basal P30518 increased in the first observation interval . Afterwards they stabilized respectively at 0.35-0.42 U/kg/24 h and 0.35-0.39 U/kg/24 h . Severe hypoglycaemia occurred 6.72/100 patient-years . CONCLUSION : P01308 therapy aimed at maintaining long-term good metabolic control is possible to achieve and is safe in young diabetic children . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . P50281 expression in first-trimester placental tissue is upregulated in type 1 diabetes as a result of elevated insulin and tumor necrosis factor-alpha levels . OBJECTIVE : In pregestational diabetes , the placenta at term of gestation is characterized by various structural and functional changes . Whether similar alterations occur in the first trimester has remained elusive . Placental development requires proper trophoblast invasion and tissue remodeling , processes involving matrix metalloproteinases ( MMPs ) of which the membrane-anchored members ( MT-MMPs ) such as MT1-MMPs are key players . Here , we hypothesize a dysregulation of placental P50281 in the first trimester of type 1 diabetic pregnancies induced by the diabetic environment . RESEARCH DESIGN AND METHODS : P50281 protein was measured in first-trimester placentas of healthy ( n = 13 ) and type 1 diabetic ( n = 13 ) women . To identify potential regulators , first-trimester trophoblasts were cultured under hyperglycemia and various insulin , P05019 , P01344 , and tumor necrosis factor-alpha ( P01375 ) concentrations in presence or absence of signaling pathway inhibitors . RESULTS : P50281 was strongly expressed in first-trimester trophoblasts . In type 1 diabetes , placental pro- P50281 was upregulated , whereas active P50281 expression was only increased in late first trimester . In isolated primary trophoblasts , insulin , P05019 , P01344 , and P01375 upregulated P50281 expression , whereas glucose had no effect . The insulin effect was dependent on phosphatidylinositol 3-kinase , the P05019 effect on mitogen-activated protein kinase , and the P01344 effect on both . CONCLUSIONS : This is the first study reporting alterations in the first-trimester placenta in type 1 diabetes . The upregulated P50281 expression in type 1 diabetes may be the result of higher maternal insulin and P01375 levels . We speculate that the elevated P50281 will affect placental development and may thus contribute to long-term structural alterations in the placenta in pregestational diabetes . Phenotypic and molecular evaluation of a chromosome 1q region with linkage and association to type 2 diabetes in humans . OBJECTIVE : Linkage to type 2 diabetes ( T2D ) is well replicated on chromosome 1q21-q23 . Within this region , T2D was associated with common single nucleotide polymorphisms that marked an extended linkage disequilibrium block , including the liver pyruvate kinase gene ( P30613 ) , in several European-derived populations . In this study we sought to determine the molecular basis for the association and the phenotypic consequences of the risk haplotype . RESEARCH DESIGN AND METHODS : Genes surrounding P30613 were resequenced in European-American and African-American cases and controls , and association with T2D was tested . Copy number variants ( CNVs ) were tested for four regions with real-time PCR . Expression of genes in the region was tested in adipose and muscle from nondiabetic subjects with each genotype . P01308 secretion , insulin sensitivity , and hepatic glucose production were tested in nondiabetic individuals with each haplotype combination . RESULTS : No coding variant in the region was associated with T2D . CNVs were rare and not associated with T2D . P30613 was not expressed in available tissues , but expression of genes Q9P1Z3 , P49760 , O14828 , and P14324 was not associated with haplotype combinations in adipose or muscle . Haplotype combinations were not associated with insulin secretion or peripheral insulin sensitivity , but homozygous carriers of the risk haplotype had increased hepatic glucose production during hyperinsulinemia . CONCLUSIONS : Noncoding variants in the P30613 region likely alter gene expression of one or more genes . Our extensive physiological and molecular studies suggest increased hepatic glucose production and reduced hepatic insulin sensitivity , thus pointing to P30613 itself as the most likely candidate gene in this population . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . Effects of I(Ks) channel inhibitors in insulin-secreting P01308 -1 cells . DB01345 channels regulate insulin secretion . The closure of K( DB00171 ) channels leads to membrane depolarisation , which triggers Ca(2+) influx and stimulates insulin secretion . The subsequent activation of K(+) channels terminates secretion . We examined whether P51787 channels are expressed in pancreatic beta-cells and analysed their functional role . Using RT/PCR cellular mRNA of P51787 but not of P15382 channels was detected in P01308 -1 cells . Effects of two sulfonamide analogues , 293B and HMR1556 , inhibitors of P51787 channels , were examined on voltage-activated outwardly rectifying K(+) currents using the patch-clamp method . It was found that 293B inhibited 60 % of whole-cell outward currents induced by voltage pulses from -70 to +50 mV with a concentration for half-maximal inhibition ( IC(50) ) of 37 microM . The other sulfonamide analogue HMR1556 inhibited 48 % of the outward current with an IC(50) of 7 microM . The chromanol 293B had no effect on tolbutamide-sensitive K( DB00171 ) channels . Action potentials induced by current injections were broadened and after-repolarisation was attenuated by 293B . P01308 secretion in the presence but not in the absence of tolbutamide was significantly increased by 293B . These results suggest that 293B- and HMR1556-sensitive channels , probably in concert with other voltage-activated K(+) channels , influence action potential duration and frequency and thus insulin secretion . Identification of insulin-stimulated phosphorylation sites on calmodulin . P01308 enhances calmodulin phosphorylation in vivo . To determine the insulin-sensitive phosphorylation sites , phosphocalmodulin was immunoprecipitated from Chinese hamster ovary cells expressing human insulin receptors ( CHO/IR ) . P62158 was constitutively phosphorylated on serine , threonine , and tyrosine residues , and insulin enhanced phosphate incorporation on serine and tyrosine residues . Phosphocalmodulin immunoprecipitated from control and insulin-treated CHO/IR cells , and calmodulin phosphorylated in vitro by the insulin receptor kinase and casein kinase II were resolved by two-dimensional phosphopeptide mapping . Several common phosphopeptides were detected . The phosphopeptides from the in vitro maps were eluted and phosphoamino acid analysis , manual sequencing , strong cation exchange chromatography , and additional proteolysis were performed . This strategy demonstrated that DB00135 -99 and DB00135 -138 were phosphorylated in vitro by the insulin receptor kinase and DB00156 -79 , DB00133 -81 , DB00133 -101 and DB00156 -117 were phosphorylated by casein kinase II . In vivo phosphorylation sites were identified by comigration of phosphopeptides on two-dimensional maps with phosphopeptides derived from calmodulin phosphorylated in vitro and by phosphoamino acid analysis . This approach revealed that DB00135 -99 and DB00135 -138 of calmodulin were phosphorylated in CHO/IR cells in response to insulin . Additional sites remain to be identified . The identification of the insulin-stimulated in vivo tyrosine phosphorylation sites should facilitate the elucidation of the physiological role of phosphocal-modulin . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Dietary soy protein isolate attenuates metabolic syndrome in rats via effects on Q07869 , LXR , and SREBP signaling . To determine the effects of feeding soy or isoflavones on lipid homeostasis in early development , weanling rats were fed AIN-93G diets made with casein , soy protein isolate ( SPI+ ) , isoflavone-reduced SPI+ ( SPI- ) , or casein supplemented with genistein or daidzein for 14 d . PPARalpha-regulated genes and proteins involved in fatty acid degradation were upregulated by SPI+ ( P < 0.05 ) accompanied by increased promoter binding and expression of PPARalpha mRNA ( P < 0.05 ) . Feeding SPI- or pure isoflavones did not alter PPARalpha-regulated pathways . SPI+ feeding had similar effects on PPARgamma signaling . SPI+ , SPI- , and casein plus isoflavones all increased liver Q9UBH6 (LXR)alpha-regulated genes and enzymes involved in cholesterol homeostasis . Feeding SPI+ increased promoter binding of LXRalpha , expression of the transcription factor mRNA , and protein ( P < 0.05 ) . In a second experiment , male Sprague-Dawley rats were fed casein diets from postnatal d ( P01160 ) 24 to PND64 or were fed high-fat Western diets containing 5 g x kg(-1) cholesterol made with either casein or SPI+ . P01308 resistance , steatosis , and hypercholesterolemia in the Western diet-fed rats were partially prevented by SPI+ ( P < 0.05 ) . Nuclear sterol receptor element binding protein ( SREBP ) -1c protein and mRNA and protein expression of enzymes involved in fatty acid synthesis were increased by feeding Western diets containing casein but not SPI+ ( P < 0.05 ) . These data suggest that activation of Q07869 and LXR signaling and inhibition of SREBP-1c signaling may contribute to insulin sensitization and improved lipid homeostasis in SPI+-fed rats after consumption of diets high in fat and cholesterol . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development .

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interacts_with DB01017?

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aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Anticancer potential of curcumin : preclinical and clinical studies . Curcumin ( diferuloylmethane ) is a polyphenol derived from the plant Curcuma longa , commonly called turmeric . Extensive research over the last 50 years has indicated this polyphenol can both prevent and treat cancer . The anticancer potential of curcumin stems from its ability to suppress proliferation of a wide variety of tumor cells , down-regulate transcription factors NF-kappa B , AP-1 and Egr-1 ; down-regulate the expression of P35354 , P28300 , NOS , P14780 , uPA , P01375 , chemokines , cell surface adhesion molecules and cyclin D1 ; down-regulate growth factor receptors ( such as P00533 and P04626 ) ; and inhibit the activity of c-Jun N-terminal kinase , protein tyrosine kinases and protein serine/threonine kinases . In several systems , curcumin has been described as a potent antioxidant and anti-inflammatory agent . Evidence has also been presented to suggest that curcumin can suppress tumor initiation , promotion and metastasis . Pharmacologically , curcumin has been found to be safe . Human clinical trials indicated no dose-limiting toxicity when administered at doses up to 10 g/day . All of these studies suggest that curcumin has enormous potential in the prevention and therapy of cancer . The current review describes in detail the data supporting these studies . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Matrix metalloproteinase ( MMP ) -9 , but not P08253 , is involved in the development and progression of C protein-induced myocarditis and subsequent dilated cardiomyopathy . Repeated or continuous inflammation of the heart is one of the initiation factors for dilated cardiomyopathy ( DCM ) . In previous studies , we established a DCM animal model by immunizing rats with cardiac C protein . In the present study , we analyze the role of matrix metalloproteinases ( MMPs ) in experimental autoimmune carditis ( EAC ) and subsequent DCM to elucidate the pathomechanisms of this disease . In this model , inflammation begins approximately 9 days after immunization . At that time , MMP activities were detected by in situ zymography . Real-time PCR analysis revealed continuous up-regulation of P08253 mRNA from 2 wk and thereafter . P14780 mRNA , however , had only a transient increase at 2 wk . Double staining with in situ zymography and cell markers demonstrated that gelatinase ( P08253 and P14780 ) -expressing cells are infiltrating macrophages during the early stage and cardiomyocytes at later stages . DB01017 , which inhibits P14780 activities more strongly than P08253 , significantly suppressed EAC , but an P08253 -specific inhibitor , TISAM , did not affect the course of the disease . Furthermore , immunohistochemical examination revealed that minocycline treatment suppressed T cell and macrophage infiltration strongly , whereas TISAM did not . These findings indicate that P14780 , but not P08253 , is involved in the pathogenesis of the acute phase of EAC , and further suggest that P14780 inhibitors , minocycline and its derivatives , may be useful therapies for EAC and DCM . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . P37840 activates microglia by inducing the expressions of matrix metalloproteinases and the subsequent activation of protease-activated receptor-1 . The mutation or overexpression of alpha-synuclein protein plays a pivotal role in the pathogenesis of Parkinson 's disease . In our preliminary experiments , we found that alpha-synuclein induced the expression of matrix metalloproteinases ( MMPs ) ( P03956 , -3 , -8 , and -9 ) in rat primary cultured microglia . Thus , the current study was undertaken to determine the roles of MMPs in alpha-synuclein-induced microglial activation . The inhibition of P08254 , -8 , or -9 significantly reduced NO and reactive oxygen species levels and suppressed the expression of P01375 and IL-1beta . Notably , P22894 inhibitor suppressed P01375 production more efficaciously than P08254 or P14780 inhibitors . Inhibition of P08254 or -9 also suppressed the activities of MAPK , NF-kappaB , and AP-1 . Previously , protease-activated receptor-1 ( P25116 ) has been associated with the actions of MMPs , and thus , we further investigated the role of P25116 in alpha-synuclein-induced inflammatory reactions . A P25116 -specific inhibitor and a P25116 antagonist significantly suppressed cytokine levels , and NO and reactive oxygen species production in alpha-synuclein-treated microglia . Subsequent P25116 cleavage assay revealed that P08254 , -8 , and -9 , but not alpha-synuclein , cleaved the synthetic peptide containing conventional P25116 cleavage sites . These results suggest that MMPs secreted by alpha-synuclein-stimulated microglia activate P25116 and amplify microglial inflammatory signals in an autocrine or paracrine manner . Furthermore , our findings suggest that modulation of the activities of MMPs and/or P25116 may provide a new therapeutic strategy for Parkinson 's disease . DB01017 and tissue-type plasminogen activator for stroke : assessment of interaction potential . BACKGROUND AND PURPOSE : New treatment strategies for acute ischemic stroke must be evaluated in the context of effective reperfusion . DB01017 is a neuroprotective agent that inhibits proteolytic enzymes and therefore could potentially both inactivate the clot lysis effect and decrease the damaging effects of tissue-type plasminogen activator ( t-PA ) . This study aimed to determine the effect of minocycline on t-PA clot lysis and t-PA-induced hemorrhage formation after ischemia . METHODS : Fibrinolytic and amidolytic activities of t-PA were investigated in vitro over a range of clinically relevant minocycline concentrations . A suture occlusion model of 3-hour temporary cerebral ischemia in rats treated with t-PA and 2 different minocycline regimens was used . Blood-brain barrier basal lamina components , matrix metalloproteinases ( MMPs ) , hemorrhage formation , infarct size , edema , and behavior outcome were assessed . RESULTS : DB01017 did not affect t-PA fibrinolysis . However , minocycline treatment at 3 mg/kg IV decreased total protein expression of both P08253 ( P=0.0034 ) and P14780 ( P=0.001 for 92 kDa and P=0.0084 for 87 kDa ) . It also decreased the incidence of hemorrhage ( P=0.019 ) , improved neurologic outcome ( P=0.0001 for Bederson score and P=0.0391 for paw grasp test ) , and appeared to decrease mortality . MMP inhibition was associated with decreased degradation in collagen IV and laminin-alpha1 ( P=0.0001 ) . CONCLUSIONS : Combination treatment with minocycline is beneficial in t-PA-treated animals and does not compromise clot lysis . These results also suggest that neurovascular protection by minocycline after stroke may involve direct protection of the blood-brain barrier during thrombolysis with t-PA . Clinical and pathogenic aspects of candidate genes for lithium prophylactic efficacy . A number of candidate genes for lithium prophylactic efficacy have been proposed , some of them being also associated with a predisposition to bipolar illness . The aim of the present study was to investigate a possible association between polymorphisms of 14 common genes with the quality of prophylactic lithium response in patients with bipolar mood disorder , in relation to the putative role of these genes in the pathogenesis of this disorder . Some association with lithium prophylactic efficacy was found for the polymorphisms of P31645 , P21728 , P21964 , P23560 and P06241 genes , but not for 5HT2A , 5HT2C , P14416 , P35462 , P21917 , GSK-3 , Q16620 , Q13224 and P14780 . Possible aspects of these genes with regard to the mechanism of lithium activity and pathogenesis of bipolar mood disorder are discussed . Increased serum levels of neutrophil gelatinase-associated lipocalin in patients with essential thrombocythemia and polycythemia vera . Neutrophil gelatinaase-associated lipocalin ( P80188 ) is a glycoprotein bound with matrix metalloproteinase-9 ( P14780 ) in human neutrophils , and elevated tissue P80188 expression has been documented in different infectious and inflammatory conditions . Recent evidence suggests that P80188 expression is induced in many types of human cancer . Moreover , P80188 is required for P11274 - P00519 -induced tumorigenesis . The aim of the present study was to measure serum levels of P80188 in patients with essential thrombocythemia ( ET ) and polycythemia vera ( PV ) . We also evaluated P80188 levels in patients with ET and PV with and without thrombotic events , to explore a possible correlation of P80188 with platelet and leukocyte activation , and in patients with sepsis . Serum P80188 levels in the study population were significantly higher than in healthy adults and in subjects with sepsis . A correlation between P80188 and the number of white cells and neutrophils was found in patients with PV and ET . P80188 serum levels were not different depending on the presence or not of the O60674 mutation , and a mutant allele dosage effect was not observed for P80188 levels . Patients with PV and ET with thrombosis did not have significantly higher levels of P80188 . We were unable to demonstrate a significant association between serum P80188 levels and CD11b or CD62 expression . In conclusion , our study reports evidence demonstrating that increased levels of P80188 appear to be a characteristic of patients with PV and ET . Downregulation of the P61073 receptor inhibits cervical carcinoma metastatic behavior in vitro and in vivo . Cervical carcinoma is frequently diagnosed among women , particularly in low and middle income countries . In this study , we investigated the role of the P48061 / P61073 axis during cervical carcinoma growth and progression in vitro and in vivo . Downregulation of P61073 receptor using an RNA interference system led to almost complete inhibition of the receptor expression , activation and function . P61073 receptor silencing led to decreased ability to signal , to induce migration and to form holoclone-like colonies , with no influence on viability/proliferation of the cells . P61073 -deficient cells had also significantly lower levels of P14780 . Interestingly , downregulation of P61073 expression resulted in reduced tumor growth in vivo . Tumors generated by P61073 -deficient cells had also lower expression of the proliferation marker Ki‑67 and decreased ability to engraft into lungs and spleen . Taken together , our results indicate that P61073 receptor may play an important role during cervical carcinoma invasion . In our study P61073 influenced invasive properties of cervical carcinoma cells both in vitro and in vivo . Depletion of Nrf2 enhances inflammation induced by oxyhemoglobin in cultured mice astrocytes . Q16236 ( Nrf2 ) -antioxidant response element pathway has been proved to be the key regulator in reducing inflammatory damage , which is involved in subarachnoid hemorrhage ( Q53FZ2 ) . Here , in a traditional in vitro Q53FZ2 model , we investigated the effect of Nrf2 depletion on pro-inflammatory cytokines production . Primary cultured astrocytes from Nrf2 wild type ( WT ) or knockout ( KO ) mouse were exposed or not exposed to oxyhemoglobin ( OxyHb ) . Then the DNA-binding activity of transcription factor nuclear factor-κB ( NF-κB ) was detected by EMSA . The expression of P01375 -α , IL-1β , P05231 and P14780 were evaluated . The activity of P14780 was measured by Gelatin zymography . After exposure to OxyHb , NF-κB was activated and the expression of downstream pro-inflammatory cytokines was up-regulated in astrocytes . And such up-regulation was much higher in KO astrocytes than in WT astrocytes , which means more aggravated inflammation in Nrf2 deficient astrocytes . These results suggest that astrocytes participate in inflammatory process after Q53FZ2 and the absence of Nrf2 may induce more aggressive inflammation through activation of NF-κB pathway . DB01017 protects against permanent cerebral ischemia in wild type but not in matrix metalloprotease-9-deficient mice . DB01017 is protective in models of transient middle cerebral artery occlusion ( MCAO ) . We studied whether minocycline and doxycycline , another tetracycline derivative , provide protection in permanent MCAO . Because minocycline inhibits matrix metalloprotease-9 ( P14780 ) , we also compared minocycline 's protective effect in wild type ( wt ) and P14780 knock-out ( ko ) mice . Wt FVB/N , Balb/C , and two lines of P14780 ko and their wt C57Bl/6 control mice were subjected to 24- or 72-hour permanent MCAO . Drug administration was started either 12 hours before or 2 hours after the onset of MCAO . Infarct size was determined by triphenyltetrazolium staining or P24752 -weighted Q9BWK5 . Zymography was used to study the expression of MMPs . In wt strains , tetracycline treatments started before MCAO reduced the infarct size by 25 % to 50 % , whereas the treatment started after MCAO was not protective . DB01017 inhibited ischemia-provoked pro- P14780 induction in wt mice , but was not protective in P14780 ko mice . Pro- P08253 was induced by MCAO in wt and P14780 ko mice . MCAO-induced pro- P08253 was downregulated by minocycline treatment in wt mice but remained in P14780 ko mice at the same level as in saline-treated wt mice . Tetracyclines are protective in permanent MCAO when the treatment is started before the insult . DB01017 may provide protection by interfering with MMPs . DB01017 attenuates hypoxia-inducible factor-1α expression correlated with modulation of p53 and AKT/ P42345 /p70S6K/ Q13541 pathway in ovarian cancer : in vitro and in vivo studies . Hypoxia-inducible factor ( HIF ) -1α is the key cellular survival protein under hypoxia , and is associated with tumor progression and angiogenesis . We have recently shown the inhibitory effects of minocycline on ovarian tumor growth correlated with attenuation of vascular endothelial growth factor ( P15692 ) and herein report a companion laboratory study to test if these effects were the result of HIF-1α inhibition . In vitro , human ovarian carcinoma cell lines ( A2780 , OVCAR-3 and SKOV-3 ) were utilized to examine the effect of minocycline on Q9BYW2 and its upstream pathway components to elucidate the underlying mechanism of action of minocycline . Mice harboring OVCAR-3 xenografts were treated with minocycline to assess the in vivo efficacy of minocycline in the context of Q9BYW2 . DB01017 negatively regulated HIF-1α protein levels in a concentration-dependent manner and induced its degradation by a mechanism that is independent of prolyl-hydroxylation . The inhibition of HIF-1α was found to be associated with up-regulation of endogenous p53 , a tumor suppressor with confirmed role in HIF-1α degradation . Further studies demonstrated that the effect of minocycline was not restricted to proteasomal degradation and that it also caused down-regulation of HIF-1α translation by suppressing the AKT/ P42345 /p70S6K/ Q13541 signaling pathway . DB01017 treatment of mice bearing established ovarian tumors , led to suppression of HIF-1α accompanied by up-regulation of p53 protein levels and inactivation of AKT/ P42345 /p70S6K/ Q13541 pathway . These data reveal the therapeutic potential of minocycline in ovarian cancer as an agent that targets the pro-oncogenic factor HIF-1α through multiple mechanisms . Rosiglitazone regulates P05231 -stimulated lipolysis in porcine adipocytes . Interleukin ( IL ) -6 , a proinflammatory cytokine , stimulates adipocyte lipolysis and induces insulin resistance in obese and diabetic subjects . However , the effects of the anti-diabetic drug rosiglitazone on P05231 -stimulated lipolysis and the underlying molecular mechanism are largely unknown . In this study , we demonstrated that rosiglitazone suppressed P05231 -stimulated lipolysis in differentiated porcine adipocytes by inactivation of extracellular signal-related kinase ( P29323 ) . Meanwhile , rosiglitazone enhanced the lipolysis response of adipocytes to isoprenaline . In addition , rosiglitazone significantly reversed P05231 -induced down-regulation of several genes such as perilipin A , peroxisome proliferators activated receptor gamma ( Q07869 & gamma ; ) , and fatty acid synthetase , as well as the up-regulation of P05231 mRNA . However , mRNA expression of Q07869 & gamma ; coactivator-1 alpha ( DB01053 -1 & alpha ; ) was enhanced by rosiglitazone in P05231 -stimulated adipocytes . These results indicate that rosiglitazone suppresses P05231 -stimulated lipolysis in porcine adipocytes through multiple molecular mechanisms . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . DB01017 promotes dendritic spine maturation and improves behavioural performance in the fragile X mouse model . BACKGROUND : Fragile X syndrome ( FXS ) is the most common single gene inherited form of mental retardation , with behaviours at the extreme of the autistic spectrum . Subjects with FXS and fragile X mental retardation gene knock out ( Fmr1 KO ) mice , an animal model for FXS , have been shown to exhibit defects in dendritic spine maturation that may underlie cognitive and behavioural abnormalities in FXS . DB01017 is a tetracycline analogue that has been used in clinical trials for stroke , multiple sclerosis and several neurodegenerative conditions . METHODS : We evaluated the effects of minocycline on dendritic spine development in the hippocampus of young Fmr1 KO mice , and in primary cultures of hippocampal neurons isolated from those mice . Cognitive effects of minocycline in young WT and Fmr1 KO mice were also evaluated using established behavioural tests for general cognition , activity and anxiety . RESULTS : Our studies demonstrate that minocycline promotes dendritic spine maturation both in cultures and in vivo . The beneficial effects of minocycline on dendritic spine morphology are also accompanied by changes in the behavioural performance of 3-week-old Fmr1 KO mice . DB01017 treated Fmr1 KO mice show less anxiety in the elevated plus maze and more strategic exploratory behaviour in the Y maze as compared to untreated Fmr1 KO mice . Our data suggest that these effects of minocycline may relate to its inhibitory action on P14780 expression and activity , which are higher in the hippocampus of Fmr1 KO mice . CONCLUSION : These findings establish minocycline as a promising therapeutic for the treatment of fragile X mental retardation . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood-lymphocytes . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood lymphocytes was investigated in vitro . The effects of the antibiotics on the mitotic response of purified lymphocytes stimulated with P01584 varied according to the individual from whom the lymphocytes were obtained . At concentrations above those reported to be present in serum during conventional therapy ( 2-8 mg/l ) , there was a tendency for both minocycline and tetracycline to suppress the mitotic response . DB01017 was superior to tetracycline in this respect . However , at physiological concentrations the antibiotics either had no significant effect , suppressed the mitotic response ( minocycline at 2 mg/l with one of six donors ) , or enhanced the mitotic response ( tetracycline at 2 and 8 mg/l with four of six donors ) . The stimulatory effect of tetracycline was not demonstrated when lymphocytes were cultured in whole blood for up to seven days with the antibiotic alone . Similar effects of the antibiotics were observed when mononuclear cell fractions isolated from six donors were stimulated with an optimal concentration of phytohaemagglutinin ( PHA ) . Stimulation of lymphocytes in whole blood cultures with PHA in the presence of minocycline and tetracycline revealed that , under these culture conditions , the antibiotics could suppress the mitotic response of lymphocytes at physiological doses with cells from a majority of donors . Glioma-associated microglial P14780 expression is upregulated by O60603 signaling and sensitive to minocycline . The invasiveness of malignant gliomas is one of the major obstacles in glioma therapy and the reason for the poor survival of patients . Glioma cells infiltrate into the brain parenchyma and thereby escape surgical resection . Glioma associated microglia/macrophages support glioma infiltration into the brain parenchyma by increased expression and activation of extracellular matrix degrading proteases such as matrix metalloprotease ( MMP ) 2 , P14780 and membrane-type 1 MMP . In this work we demonstrate that , P14780 is predominantly expressed by glioma associated microglia/macrophages in mouse and human glioma tissue but not by the glioma cells . Supernatant from glioma cells induced the expression of P14780 in cultured microglial cells . Using mice deficient for different Toll-like receptors we identified O60603 /6 as the signaling pathway for the glioma induced upregulation of microglial P14780 . Also in an experimental mouse glioma model , O60603 deficiency attenuated the upregulation of microglial P14780 . Moreover , glioma supernatant triggered an upregulation of O60603 expression in microglia . Both , the upregulation of P14780 and O60603 were attenuated by the antibiotic minocycline and a p38 mitogen-activated protein kinase antagonist in vitro . DB01017 also extended the survival rate of glioma bearing mice when given to the drinking water . Thus glioma cells change the phenotype of glioma associated microglia/macrophages in a complex fashion using O60603 as an important signaling pathway and minocycline further proved to be a potential candidate for adjuvant glioma therapy . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . DB01017 inhibits contusion-triggered mitochondrial cytochrome c release and mitigates functional deficits after spinal cord injury . We investigated whether permeability transition-mediated release of mitochondrial cytochrome c is a potential therapeutic target for treating acute spinal cord injury ( SCI ) . Based on previous reports , minocycline , a second-generation tetracycline , exerts neuroprotection partially by inhibiting mitochondrial cytochrome c release and reactive microgliosis . We first evaluated cytochrome c release at the injury epicenter after a P28907 contusive SCI in rats . P99999 release peaked at approximately 4-8 h postinjury . A dose-response study generated a safe pharmacological regimen that enabled i.p. minocycline to significantly lower cytosolic cytochrome c at the epicenter 4 h after SCI . In the long-term study , i.p. minocycline ( 90 mg/kg administered 1 h after SCI followed by 45 mg/kg administered every 12 h for 5 days ) markedly enhanced long-term hind limb locomotion relative to that of controls . Coordinated motor function and hind limb reflex recoveries also were improved significantly . Histopathology suggested that minocycline treatment alleviated later-phase tissue loss , with significant sparing of white matter and ventral horn motoneurons at levels adjacent to the epicenter . Furthermore , glial fibrillary acidic protein and 2',3' cyclic nucleotide 3' phosphodiesterase immunocytochemistry showed an evident reduction in astrogliosis and enhanced survival of oligodendrocytes . Therefore , release of mitochondrial cytochrome c is an important secondary injury mechanism in SCI . Drugs with multifaceted effects in antagonizing this process and microgliosis may protect a proportion of spinal cord tissue that is clinically significant for functional recovery . DB01017 , with its proven clinical safety , capability to cross the blood-brain barrier , and demonstrated efficacy during a clinically relevant therapeutic window , may become an effective therapy for acute SCI . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Antenatal maternal long-term hypoxia : acclimatization responses with altered gene expression in ovine fetal carotid arteries . In humans and other species , long-term hypoxia ( LTH ) during pregnancy can lead to intrauterine growth restriction with reduced body/brain weight , dysregulation of cerebral blood flow ( Q03701 ) , and other problems . To identify the signal transduction pathways and critical molecules , which may be involved in acclimatization to high altitude LTH , we conducted microarray with advanced bioinformatic analysis on carotid arteries ( CA ) from the normoxic near-term ovine fetus at sea-level and those acclimatized to high altitude for 110+ days during gestation . In response to LTH acclimatization , in fetal CA we identified mRNA from 38 genes upregulated > 2 fold ( P < 0.05 ) and 9 genes downregulated > 2-fold ( P < 0.05 ) . The major genes with upregulated mRNA were P43003 , P01308 -like growth factor ( IGF ) binding protein 3 , IGF type 2 receptor , transforming growth factor ( TGF ) Beta-3 , and genes involved in the AKT and P10415 signal transduction networks . Most genes with upregulated mRNA have a common motif for Pbx/Knotted homeobox in the promoter region , and Sox family binding sites in the 3' un translated region ( UTR ) . Genes with downregulated mRNA included those involved in the P04637 pathway and P09917 activating proteins . The promoter region of all genes with downregulated mRNA , had a common 49 bp region with a binding site for DOT6 and TOD6 , components of the RPD3 histone deacetylase complex RPD3C(L) . We also identified miRNA complementary to a number of the altered genes . Thus , the present study identified molecules in the ovine fetus , which may play a role in the acclimatization response to high-altitude associated LTH . DB01017 inhibits P09917 expression and accelerates functional recovery in chronic phase of focal cerebral ischemia in rats . AIMS : We previously reported that minocycline attenuates acute brain injury and inflammation after focal cerebral ischemia , and this is partly mediated by inhibition of P09917 ( 5- P28300 ) expression . Here , we determined the protective effect of minocycline on chronic ischemic brain injury and its relation with the inhibition of 5- P28300 expression after focal cerebral ischemia . MAIN METHODS : Focal cerebral ischemia was induced by 90 min of middle cerebral artery occlusion followed by reperfusion for 36 days . DB01017 ( 45 mg/kg ) was administered intraperitoneally 2h and 12h after ischemia and then every 12h for 5 days . Sensorimotor function was evaluated 1-28 days after ischemia and cognitive function was determined 30-35 days after ischemia . Thereafter , infarct volume , neuron density , astrogliosis , and 5- P28300 expression in the brain were determined . KEY FINDINGS : DB01017 accelerated the recovery of sensorimotor and cognitive functions , attenuated the loss of neuron density , and inhibited astrogliosis in the boundary zone around the ischemic core , but did not affect infarct volume . DB01017 significantly inhibited the increased 5- P28300 expression in the proliferated astrocytes in the boundary zone , and in the macrophages/microglia in the ischemic core . SIGNIFICANCE : DB01017 accelerates functional recovery in the chronic phase of focal cerebral ischemia , which may be partly associated with the reduction of 5- P28300 expression . DB01017 inhibits malignant ascites of ovarian cancer through targeting multiple signaling pathways . OBJECTIVES : To evaluate the effect of minocycline on the expression of cytokines and growth factors responsible for malignant ascite formation . METHODS : In vitro , cells obtained from malignant ascites were pre-treated with minocycline ( 0-100 μmol/L ) and exposed briefly to hypoxia . In vivo , female nude mice bearing OVCAR-3 tumors were treated orally in drinking water with minocycline for 4 weeks . Plasma , ascites , and tumors were analyzed . RESULTS : DB01017 blocked hypoxia-induced surge in interleukin-6 ( P05231 ) , its soluble receptor ( sIL-6R ) and vascular endothelial growth factor ( P15692 ) levels in concentration-dependent manner . In mice , orally administered minocycline led to dramatic reduction in tumor weight and malignant ascite volume . P05231 , sIL6R and in particular P15692 levels were highly suppressed in plasma , ascite fluid and tumor tissue by minocycline . In addition , tumors from minocycline treated mice expressed profoundly lower levels of phosphorylated extracellular regulated kinases ( p-Erk1/2 ) and p-Akt . DB01017 was also effective at suppressing transforming growth factor beta ( TGF-β1 ) and increasing vascular endothelial cadherin ( P33151 ) expression thereby providing molecular confirmation for its effects on malignant ascite formation . CONCLUSION : Orally administered minocycline is highly effective in suppressing ovarian cancer-induced malignant ascites by targeting cytokines and growth factors essential for tumor growth and malignant ascite formation . P04626 mRNA status contributes to the discrepancy between gene amplification and protein overexpression in gastric cancer . BACKGROUND : Human epidermal growth factor receptor 2 ( P04626 ) is an important proto-oncogene of prognostic use in gastric cancer ( GC ) . Fluorescence in-situ hybridization ( Q5TCZ1 ) and immunohistochemistry ( IHC ) are the main clinical methods of detection of P04626 , but consistency between the methods is poor and the cause of the discrepancy is unclear . AIM : To investigate the involvement of P04626 mRNA status in the disparity between gene amplification and protein overexpression . METHODS : We investigated P04626 gene , mRNA , and protein profiles in gastric precancer and cancer tissues by use of the molecular approaches Q5TCZ1 , real-time polymerase chain reaction , and IHC . The relationships between P04626 and matrix metalloproteinase 9 ( P14780 ) and O15105 expression were analyzed and the involvement of P04626 in the interaction between tumor cells and lymphocytes was investigated by coculturing GC cell lines with peripheral blood mononuclear cells ( PBMCs ) . RESULTS : P04626 protein expression was significantly increased in cancer compared with precancer ( P = 0.003 ) , and the corresponding mRNA levels were significantly lower in precancer and cancer tissues than in normal tissues ( κ = 0.290 , P = 0.025 ) . P04626 mRNA levels were significantly higher in tumor than in peritumor tissue ( P = 0.028 ) , and were positively correlated with P14780 and O15105 mRNA levels in tumor tissues . P04626 mRNA expression in GC cell lines was increased by coculture with PBMCs . CONCLUSIONS : Different P04626 mRNA profiles , possibly in relation to contact between tumor cells and lymphocytes , might help to explain the discrepancy between gene amplification and protein overexpression results . Delayed cardioprotective effects of WY-14643 are associated with inhibition of P08253 and modulation of Bcl-2 family proteins through Q07869 -α activation in rat hearts subjected to global ischaemia-reperfusion . Peroxisome proliferator-activated receptors ( PPARs ) are ligand-activated transcription factors regulating cardiac lipid metabolism and energy homeostasis . Although the activation of PPARs has been implicated in cardioprotection , the molecular mechanisms are largely unexplored . In this study , we aimed to investigate the effect of the Q07869 -α agonist WY-14643 ( WY ) , mimicking a delayed effect of preconditioning in rat hearts exposed to acute ischaemia-reperfusion ( I/R ) 24 h later , and to define whether antioxidative and antiapoptotic mechanisms are involved . Treatment with WY markedly attenuated post-ischaemic contractile dysfunction ( as evidenced by the reduced infarct size ) , the higher left ventricular developed pressure ( LVDP ) recovery , and the decreased occurrence of arrhythmias . These effects were abolished in the presence of the Q07869 -α antagonist MK886 . Heme oxygenase-1 , a key antioxidative enzyme implicated in cytoprotection , was upregulated in response to WY at baseline , but was markedly reduced after I/R , indicating reduced oxidative stress . WY treatment was also associated with decreased mRNA levels and enzymatic activity of matrix metalloproteinase-2 , and increased ratios of Bcl-2:Bax proteins . These results indicate that Q07869 -α activation by its selective ligand WY may confer delayed preconditioning-like protection in rat hearts subjected to I/R by modulating oxidative stress , activation of matrix metalloproteinase-2 , and expression of Bcl-2 and Bax . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . DB01017 reduces renal microvascular leakage in a rat model of ischemic renal injury . Tetracyclines exhibit significant anti-inflammatory properties , inhibit matrix metalloproteinases ( MMPs ) , and are protective in models of ischemia-reperfusion injury ( IRI ) . Both inflammatory cascades and MMP activation have been demonstrated to modulate microvascular permeability . Because increased microvascular permeability occurs during IRI in a variety of organ systems including the kidney , we hypothesized that minocycline , a semisynthetic tetracycline , would diminish microvascular leakage during renal IRI . To test this hypothesis , we used intravital 2-photon microscopy to examine leakage of fluorescent dextrans from the vasculature in a rodent model of IRI . DB01017 significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Although minocycline diminished leukocyte accumulation in the kidney following ischemia , areas of leukocyte accumulation did not correlate with areas of microvascular permeability in either the saline- or minocycline-pretreated animals . DB01017 diminished the perivascular increase in P08253 and P14780 , as well as the increase in P08253 activity 24 h after ischemia . ABT-518 , a specific inhibitor of P08253 and P14780 , also significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Our results indicate that minocycline mitigates the renal microvascular permeability defect following IRI . This effect is spatially distinct from the effect of minocycline on leukocyte accumulation and may be related to diminished activity of MMPs on the integrity of the perivascular matrix . Polymorphisms of dopamine receptor/transporter genes and risk of non-small cell lung cancer . BACKGROUND : The dopaminergic pathway may be of interest in assessing risk of non-small cell lung cancer ( NSCLC ) . Dopamine receptors are expressed in alveolar epithelial cells and human lung tumours , and dopamine inhibits both cell proliferation in vitro and growth of lung tumour xenografts in nude mice . Moreover , dopamine selectively inhibits the vascular permeability and angiogenic activity of vascular endothelial growth factor ( P15692 / P15692 ) . The bioavailability of dopamine is regulated by dopamine receptors D2 ( P14416 ) , D4 ( P21917 ) and dopamine transporter 1 ( Q01959 / Q01959 ) genes . METHODS : We have analysed 10 single nucleotide polymorphisms in P14416 , P21917 and Q01959 / Q01959 genes in relation to lung cancer risk in a case-control study of smoking subjects . The study subjects were 413 healthy individuals from general population and 335 NSCLC cases . Both cases and controls were Caucasians of Norwegian origin . RESULTS : We demonstrate that P14416 polymorphisms -141Cdel , 3208G > T , TaqIB ; P21917 -521C > T and Q01959 / Q01959 -1476T > G are associated with a two- to five-fold increased NSCLC risk . The variant alleles of P14416 1412A > G and 960C > G had protective effects . CONCLUSION : The dopamine receptor/transport gene polymorphisms are associated with the risk of NSCLC among smokers . The data show that the polymorphisms resulting in lower dopamine bioavailability were associated with increased risk of NSCLC . DB01095 inhibits growth and alters the malignant phenotype of the P13671 glioma cell line . BACKGROUND : DB01095 is a member of the family of P04035 inhibitors ( statins ) extensively used in medical practice . Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development . The aim of the present study was to investigate the anti-cancer potential of fluvastatin in P13671 rat malignant glioma cells . METHODS : First , the effects of fluvastatin on cell viability ( MTT assay ) , proliferation ( BrdU assay ) , cell morphology , and cytoskeleton were examined . Subsequently , its effect on extracellular signal regulated kinase 1 and 2 ( P27361 /2 ) and P45983 and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 and P15692 was determined using a wound-healing assay and ELISA test , respectively . RESULTS : The results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor P13671 cells ( IC(50) = 8.6 μM , 48 h ) , but did not inhibit the growth of normal neuronal cells . The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm , chromatin condensation , and nucleus breakdown . CONCLUSION : The inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p- P27361 /2 expression , upregulation of p- P45983 /2 , and reduction in the P14780 and P15692 concentrations in culture media . The high anticancer ( antiproliferative , proapoptotic , antiinvasive ) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma . DB01017 suppresses experimental autoimmune encephalomyelitis by increasing tissue inhibitors of metalloproteinases . Matrix metalloproteinases ( MMPs ) that are secreted by activated T cells play a significant role in degradation of the extracellular matrix around the blood vessels and facilitate autoimmune neuroinflammation ; however , it remains unclear how MMPs act in lesion formation and whether MMP-targeted therapies are effective in disease suppression . In the present study , we attempted to treat experimental autoimmune encephalomyelitis ( EAE ) by administration of small interfering RNAs ( siRNAs ) for P08253 , P14780 , and minocycline , all of which have MMP-inhibiting functions . DB01017 , but not siRNAs , significantly suppressed disease development . In situ zymography revealed that gelatinase activities were almost completely suppressed in the spinal cords of minocycline-treated animals , while significant gelatinase activities were measured in the EAE lesions of control animals . However , P08253 and P14780 mRNAs and proteins in the spinal cords of treated rats were unexpectedly upregulated . At the same time , mRNA for tissue inhibitors of MMPs ( P01033 ) -1 and -2 were also upregulated . The EnzChek Gelatinase/Collagenase assay using tissue containing native MMPs and TIMPs demonstrated that gelatinase activity levels in the spinal cords of treated rats were suppressed to the same level as those in normal spinal cord tissues . Finally , double immunofluorescent staining demonstrated that P14780 immunoreactivities of treated rats were almost the same as those of control rats and that P14780 and P01033 immunoreactivities were colocalized in the spinal cord . These findings suggest that minocycline administration does not suppress MMPs at mRNA and protein levels but that it suppresses gelatinase activities by upregulating TIMPs . Thus , MMP-targeted therapies should be designed after the mechanisms of candidate drugs have been considered . Cardiac uptake of minocycline and mechanisms for in vivo cardioprotection . OBJECTIVES : The ability of minocycline to be transported into cardiac cells , concentrate in normal and ischemic myocardium , and act as a cardioprotector in vivo was examined . We also determined minocycline 's capacity to act as a reducer of myocardial oxidative stress and matrix metalloproteinase ( MMP ) activity . BACKGROUND : The identification of compounds with the potential to reduce myocardial ischemic injury is of great interest . Tetracyclines are antibiotics with pleiotropic cytoprotective properties that accumulate in normal and diseased tissues . DB01017 is highly lipophilic and has shown promise as a possible cardioprotector . However , minocycline 's potential as an in vivo cardioprotector as well as the means by which this action is attained are not well understood . METHODS : Rats were subjected to 45 min of ischemia and 48 h of reperfusion . Animals were treated 48 h before and 48 h after thoracotomy with either vehicle or 50 mg/kg/day minocycline . Tissue samples were used for biochemical assays and cultured cardiac cells for minocycline uptake experiments . RESULTS : DB01017 significantly reduced infarct size ( approximately 33 % ) , tissue P14780 activity , and oxidative stress . DB01017 was concentrated approximately 24-fold in normal ( 0.5 mmol/l ) and approximately 50-fold in ischemic regions ( 1.1 mmol/l ) versus blood . Neonatal rat cardiac fibroblasts , myocytes , and adult fibroblasts demonstrated a time- and temperature-dependent uptake of minocycline to levels that approximate those of normal myocardium . CONCLUSIONS : Given the high intracellular levels observed and results from the assessment of in vitro antioxidant and MMP inhibitor capacities , it is likely that minocycline acts to limit myocardial ischemic injury via mass action effects . P01375 -α-accelerated degradation of type I collagen in human skin is associated with elevated matrix metalloproteinase ( MMP ) -1 and P08254 ex vivo . P01375 ( P01375 ) -α induces matrix metalloproteinases ( MMPs ) that may disrupt skin integrity . We have investigated the effects and mechanisms of exogenous P01375 -α on collagen degradation by incubating human skin explants in defined serum-free media with or without P01375 -α ( 10ng/ml ) in the absence or presence of the nonselective MMP inhibitor DB02255 for 8 days . The basal culture conditions promoted type I collagen catabolism that was accelerated by P01375 -α ( p < 0.005 ) and accomplished by MMPs ( p < 0.005 ) . Levels of the collagenases P22894 and P45452 were insignificant and neither P08253 nor P50281 were associated with increased collagen degradation . P01375 -α increased secretion of P03956 ( p < 0.01 ) but had no impact on P03956 quantities in the tissue . Immunohistochemical analysis confirmed similar tissue P03956 expression with or without P01375 -α with epidermis being the major source of P03956 . Increased tissue-derived collagenolytic activity with P01375 -α exposure was blocked by neutralizing P03956 monoclonal antibody and was not due to down-regulation of tissue inhibitor of metalloproteinase-1 . P01375 -α increased production ( p < 0.01 ) , tissue levels ( p < 0.005 ) and catalytic activity of the endogenous P03956 activator P08254 . Type I collagen degradation correlated with P08254 tissue levels ( rs=0.68 , p < 0.05 ) and was attenuated with selective P08254 inhibitor . Type I collagen formation was down-regulated in cultured compared with native skin explants but was not reduced further by P01375 -α . P01375 -α had no significant effect on epidermal apoptosis . Our data indicate that P01375 -α augments collagenolytic activity of P03956 , possibly through up-regulation of P08254 leading to gradual loss of type I collagen in human skin . Modulation of P00533 and ROS induced cytochrome c release by combination of photodynamic therapy and carboplatin in human cultured head and neck cancer cells and tumor xenograft in nude mice . Photodynamic therapy in combination with different treatment modalities has been evaluated to study the mechanism of cellular cytotoxicity and apoptosis in various forms of cancer . In the present study , human head and neck cancer cells were treated with radachlorin mediated photodynamic therapy and the chemotherapy drug , carboplatin singly or in combination . Several parameters were studied to check the enhanced cytotoxicity of combination therapy at different time interval . From the cell viability study by MTT assay , a 22 % decrease in cell viability was observed in combination treatment . This enhanced activity of combination treatment was confirmed by cell migration assay and Hoechst PI staining . Generation of reactive oxygen species was observed and found to be higher than that of individual treatments . P99999 was found to be released from mitochondria that also induced the enhance efficacy in combination treatment . The expression of other proteins like P00533 and PARP was also modulated with the time of incubation after treatment . In the tumor xenograft study in nude mouse model , the carboplatin treated group did not show any noticeable changes in tumor volume whereas tumor volume was reduced in PDT and the combination group . Though the difference in the reduction of the tumor size was not significant between PDT and combination group , there was a difference in the expression of P00533 between these two groups . Histologic study of the inhibition in tumor growth was also performed . Therefore , this study may provide an avenue of combating head and neck cancer by a combination of conventional chemotherapy and PDT . DB01017 protects PC12 cells against DB01221 -induced injury via inhibiting P09917 activation . Recently , we have reported that minocycline , a semi-synthetic tetracycline with neuroprotective effects , inhibits the in vitro ischemic-like injury and P09917 ( 5- P28300 ) activation in PC12 cells . In the present study , we further determined whether minocycline protects PC12 cells from excitotoxicity via inhibiting 5- P28300 activation . We used N-methyl-d-aspartate ( DB01221 , 200 microM ) to induce early ( exposure for 6 h ) and delayed ( exposure for 6 h followed by 24 h recovery ) injuries . We found that DB01221 receptor antagonist ketamine , 5- P28300 inhibitor caffeic acid and minocycline concentration dependently attenuated DB01221 -induced early and delayed cell injuries ( viability reduction and cell death ) . However , only ketamine ( 1 microM ) inhibited DB01221 -evoked elevation of intracellular calcium . In addition , immunohistochemical analysis showed that DB01221 induced 5- P28300 translocation to the nuclear membrane after 1- to 6-h exposure which was confirmed by Western blotting , indicating that 5- P28300 was activated . DB01221 , caffeic acid and minocycline ( each at 1 microM ) inhibited 5- P28300 translocation after early injury . After delayed injury , PC12 cells were shrunk , and 5- P28300 was translocated to the nuclei and nuclear membrane ; ketamine , caffeic acid and minocycline inhibited both cell shrinking and 5- P28300 translocation . As a control , P16050 inhibitor baicalein showed a weak effect on cell viability and death , but no effect on 5- P28300 translocation . Therefore , we conclude that the protective effect of minocycline on DB01221 -induced injury is partly mediated by inhibiting 5- P28300 activation . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Effects of minocycline on cocaine sensitization and phosphorylation of GluR1 receptors in P09917 deficient mice . In wild-type ( WT ) mice , the antibiotic minocycline inhibits development of cocaine-induced locomotor sensitization . Some of the actions of minocycline may involve the P09917 ( 5- P28300 ) pathway . We used the model of 5- P28300 -deficient mice to investigate whether 5- P28300 participates in minocycline 's influence on the effects of cocaine . Locomotor sensitization was induced by 4 daily cocaine injections and the phosphorylation status of GluR1 glutamate receptors was assayed in brain samples . DB01017 failed to affect cocaine sensitization in 5- P28300 -deficient mice . In these mice , neither cocaine nor minocycline 4-day treatment altered GluR1 phosphorylation . In WT mice in which minocycline inhibited development of cocaine sensitization , a 4-day cocaine treatment increased GluR1 phosphorylation at both Ser831 and Ser845 sites in the frontal cortex but not the striatum ; further , this effect was prevented by minocycline . Under basal conditions and in response to a single cocaine injection the levels of GluR1 , GluR2 , and GluR3 AMPA receptor subunits did not differ between WT and 5- P28300 -deficient mice , but the response of GluR1 phosphorylation to a single cocaine injection was greater under the 5- P28300 deficiency . Hence , in WT mice GluR1 phosphorylation increased only in the frontal cortex and only at the Ser831 site . In 5- P28300 -deficient mice , acute cocaine injection increased both Ser831 and Ser845 phosphorylation both in the frontal cortex and in the striatum . We suggest that in studying minocycline 's action on cocaine 's effects and/or addiction in humans , it would be important to consider the characterization of the subjects ' 5- P28300 system . This article is part of a Special Issue entitled ' Trends in neuropharmacology : in memory of Erminio Costa ' . DB01017 partially inhibits caspase-3 activation and photoreceptor degeneration after photic injury . PURPOSE : To evaluate the possible role of caspase-3 in retinal photic injury , and to investigate whether minocycline can ameliorate light-induced photoreceptor degeneration . METHODS : Retinal photic injury was induced in rats by exposure to intense light . Expression of caspase-3 was studied using Western blot analysis , immunohistochemical staining and enzyme activity assay . Apoptotic photoreceptor cells were detected by the TdT-dUTP terminal nick-end labeling ( TUNEL ) method . DB01017 ( 15 , 30 or 45 mg/kg ) was administered before or after photic injury in rats randomly assigned to pretreatment and posttreatment groups . DB01017 and vehicle-treated retinas subjected to photic injury were compared with respect to Western blotting , enzyme activity assay , quantitative counts of TUNEL stains , morphometry of the outer nuclear layer ( ONL ) thickness and histopathological examination . RESULTS : After light exposure , active caspase-3 and poly-adenosine diphosphate-ribose-polymerase were upregulated in the retinas and increased caspase-3 immunoreactivity was observed in the ONL . P42574 enzyme activity increased in the retinas that underwent photic injury , and this increase was significantly reduced in minocycline pretreated ( 30 and 45 mg/kg ) and posttreated ( 45 mg/kg ) groups . Intraperitoneal administration of minocycline before or after photic injury in rats also resulted in less TUNEL-positive photoreceptors , as assessed by the quantitative TUNEL counts . The degree of retinal degeneration , measured by the ONL thickness 14 days after photic injury , was significantly improved in minocycline pretreatment ( 45 mg/kg ) rats . CONCLUSIONS : We demonstrate that increased caspase-3 activities localize specifically within the ONL after photic injury , and that minocycline partially inhibits caspase-3 activation and photoreceptor degeneration in this animal model . Chemotactic and cytotoxic effects of minocycline on human retinal pigment epithelial cells . PURPOSE : To reveal the effects of minocycline , an anti-inflammatory and neuroprotective agent , on the viability and physiological properties of retinal pigment epithelial ( Q96AT9 ) cells and to compare the effects with those of triamcinolone acetonide . METHODS : The proliferation of human Q96AT9 cells in vitro was investigated with a bromodeoxyuridine immunoassay ; chemotaxis was examined with a Boyden chamber assay . Cell viability was determined by trypan blue exclusion . The gene expression of growth factors and P14780 was determined with real-time RT-PCR , and the secretion of P15692 was examined with ELISA . The phosphorylation of p38 MAPK and P27361 /2 proteins was determined with Western blot analysis . RESULTS : DB01017 at low concentrations ( 50 nM-20 microM ) stimulated chemotaxis and decreased the proliferation of Q96AT9 cells . DB01017 at high concentrations ( above 5 microM ) decreased the viability of Q96AT9 cells through the induction of cell necrosis . The chemotactic effect of minocycline was mediated by the stimulation of autocrine PDGF signaling and the activation of p38 MAPK . DB01017 promoted the expression of PDGF-B , P14210 , P15692 , and P14780 and increased the amounts of phosphorylated p38 and P27361 /2 proteins in Q96AT9 cells . DB00620 reduced PDGF-evoked chemotaxis and P15692 expression and secretion and had no significant effects on cell viability and proliferation . DB00620 did not reverse the effects of minocycline on cell proliferation , chemotaxis , or viability or the expression of P15692 . CONCLUSIONS : Low-dose minocycline induces the activation of Q96AT9 cells , as indicated by the activation of p38 and P27361 /2 and by enhanced chemotaxis mediated by autocrine PDGF signaling . High-dose minocycline induces Q96AT9 cell degeneration . Inhibitors of arachidonic acid metabolism reduce DNA and nuclear fragmentation induced by P01375 plus cycloheximide in U937 cells . U937 human myeloid leukemia cells are induced to apoptosis by tumour necrosis factor ( P01375 ) plus cycloheximide ( CHX ) . We have analysed the effect of various inhibitors of the arachidonic acid ( AA ) metabolism on several features of this process . The formation of high molecular weight and oligonucleosomal DNA fragments as well as nuclear fragmentation were reduced by inhibitors of P09917 ( BWA4C and BWB70C ) , P09917 activating protein ( MK-886 ) , and cytosolic P04054 ( AACOCF3 ) . None of these agents blocked the morphological changes detected by microscopy or flow cytometry , phosphatidylserine exposure on the cell surface or Caspase 3-like activation . AA also induced nuclear fragmentation at a concentration of 1-20 microM . However , the mechanisms by which these inhibitors act , remain unexplained since there was no P09917 expression in the U937 cells and no AA release followed their stimulation with P01375 plus CHX . DB01017 inhibits angiogenesis in vitro through the translational suppression of HIF-1α . DB01017 was recently found to be effective against cancer . However , the precise molecular mechanisms of minocycline in cancer are poorly understood . Hypoxia-inducible factor-1 ( Q9BYW2 , a heterodimeric transcription factor composed of HIF-1α and β ) activates the transcription of genes that are involved in angiogenesis in cancer . In this study , we found that minocycline significantly inhibits HIF-1α protein expression and suppresses Q9BYW2 transcriptional activity . The tube formation assay showed that minocycline has anti-angiogenic activity and suppresses hypoxia-induced vascular endothelial growth factor ( P15692 ) expression . The metabolic labeling assay showed that minocycline reduces HIF-1α protein translation and global protein synthesis . In addition , minocycline suppresses P42345 signaling and increases the phosphorylation of eIF2α , which is known to be related to the translational regulation of HIF-1α expression . These findings collectively indicate that minocycline is a potential inhibitor of HIF-1α and provide new insight into the discovery of drugs for cancer treatment . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Sex-independent neuroprotection with minocycline after experimental thromboembolic stroke . BACKGROUND : DB01017 provides neurovascular protection reducing acute cerebral injury . However , it is unclear whether minocycline is effective in females . We tested minocycline in both sexes and aged animals using a novel embolic stroke model in mice that closely mimics acute thromboembolic stroke in humans . METHODS : Five groups of mice were subjected to thromboembolic stroke : adult males , aged males , adult females , aged females , and adult ovariectomized females . They were treated with phosphate saline ( vehicle ) or minocycline ( 6 mg/kg ) immediately after stroke onset . Behavioral outcomes , infarct volumes and cerebral blood flow were assessed . The effect of minocycline on expression and activity of P14780 was analyzed . RESULTS : The model resulted in reproducible infarct in the experimental groups . As expected , adult females were significantly more resistant to cerebral ischemic injury than males . This advantage was abolished by aging and ovariectomy . DB01017 significantly reduced the infarct volume ( P < 0.0001 ) and also improved neurologic score ( P < 0.0001 ) in all groups . Moreover , minocycline treatment significantly reduced mortality at 24 hours post stroke ( P = 0.037 ) for aged mice ( 25 % versus 54 % ) . Stroke up-regulated P14780 level in the brain , and acute minocycline treatment reduced its expression in both genders ( P < 0.0001 ) . CONCLUSION : In a thromboembolic stroke model minocycline is neuroprotective irrespective of mouse sex and age . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Delivering minocycline into brain endothelial cells with liposome-based technology . DB01017 has been proposed as a way to blunt neurovascular injury from matrix metalloproteinases ( MMPs ) during stroke . However , recent clinical trials suggest that high levels of minocycline may have deleterious side-effects . Here , we showed that very high minocycline concentrations damage endothelial cells via calpain/caspase pathways . To alleviate this potential cytotoxicity , we encapsulated minocycline in liposomes . Low concentrations of minocycline could not reduce tumor necrosis factor α ( TNFα ) -induced P14780 release from endothelial cells . But low concentrations of minocycline-loaded liposomes significantly reduced TNFα-induced P14780 release . This study provides proof-of-concept that liposomes may be used to deliver lower levels of minocycline for targeting MMPs in cerebral endothelium . Antiarthritic mechanisms of amyrin triterpenes . The triterpenes , alpha-amyrin ( AA ) and its palmitate ( P08697 ) and linoleate esters ( AAL ) , were tested on models of inflammatory and destructive arthritic processes and their effects were compared with the clinical antiarthritic drugs indomethacin ( IN ) and methotrexate ( MTX ) . The triterpenes had no effect on the prostaglandin phase of carrageenin pedal edema in rats , which was reduced 28 % by 100 microM IN . AAL caused a considerable reduction in the synthesis by human neutrophils of P09917 products -- 5-HETE ( IC50 = 70 microM ) , LTB4 , ( 62 microM ) , isomer I ( 30 microM ) and isomer II ( 24 microM ) . Rat osteosarcoma cell growth was inhibited by all triterpenes with IC50 's ( microM ) of < 10 ( P08697 ) , 14 ( AA ) and 27 ( AAL ) and were more effective than IN ( 35 ) . MTX caused 100 % inhibition at a concentration of 10 microM compared with 64 % inhibition by P08697 . Tadpole collagenase digestion of type I ( bone ) native collagen was completely inhibited by all the triterpenes as well as IN and MTX at 100 microM . The results indicate that the principal point of antiarthritic intervention by amyrin triterpenes lies in their local inhibition of joint destruction . Dopamine receptor D2 gene is associated with weight gain in schizophrenic patients under long-term atypical antipsychotic treatment . OBJECTIVE : Schizophrenic patients treated with atypical antipsychotics ( AAPs ) often develop excessive body weight gain ( BWG ) , which may lead to further morbidity and poor treatment compliance . This study examined whether genetic variants in the dopamine receptor D2 ( P14416 ) gene may be associated with body weight change after P08697 treatment . METHODS : The study included 479 schizophrenic patients treated with clozapine ( n=239 ) , olanzapine ( n=70 ) or risperidone ( n=170 ) for an average of 48.2+/-27.8 months . BWG was defined as an increase of more than 7 % of the baseline body weight during P08697 treatment . Thirteen common single nucleotide polymorphisms of the P14416 gene were chosen as tagging single nucleotide polymorphisms . RESULTS : In single-marker-based analysis , the P14416 rs4436578-C homozygous genotype was found to be associated with a significantly increased risk of BWG [ P=0.001 , adjusted odds ratio=3.36 ( 95 % confidence interval=1.62 - 7.00 ) ] . In addition , haplotype analysis further showed that the rs4436578-C-allele-related haplotype was more frequent in those patients with BWG than those without ( P=0.01 - 0.00019 ) . CONCLUSION : Our findings confirm the importance of genetic factors in body weight change induced by long-term P08697 treatment in patients with schizophrenia and indicate a role of P14416 in body weight regulation during long-term P08697 treatment . Involvement of pro- and antinociceptive factors in minocycline analgesia in rat neuropathic pain model . In neuropathic pain the repeated minocycline treatment inhibited the mRNA and protein expression of the microglial markers and metalloproteinase-9 ( P14780 ) . The minocycline diminished the pronociceptive ( P05231 , Q14116 ) , but not antinociceptive ( IL-1alpha , P05112 , P22301 ) cytokines at the spinal cord level . In vitro primary cell culture studies have shown that P14780 , P01033 , IL-1beta , IL-1alpha , P05231 , P22301 , and Q14116 are of microglial origin . DB01017 reduces the production of pronociceptive factors , resulting in a more potent antinociceptive effect . This change in the ratio between pro- and antinociceptive factors , in favour of the latter may be the mechanism of minocycline analgesia in neuropathy . PEG minocycline-liposomes ameliorate CNS autoimmune disease . BACKGROUND : DB01017 is an oral tetracycline derivative with good bioavailability in the central nervous system ( CNS ) . DB01017 , a potent inhibitor of matrix metalloproteinase ( MMP ) -9 , attenuates disease activity in experimental autoimmune encephalomyelitis ( EAE ) , an animal model of multiple sclerosis ( MS ) . Potential adverse effects associated with long-term daily minocycline therapy in human patients are concerning . Here , we investigated whether less frequent treatment with long-circulating polyethylene glycol ( PEG ) minocycline liposomes are effective in treating EAE . FINDINGS : Performing in vitro time kinetic studies of PEG minocycline-liposomes in human peripheral blood mononuclear cells ( PBMCs ) , we determined that PEG minocycline-liposome preparations stabilized with CaCl(2) are effective in diminishing P14780 activity . Intravenous injections of PEG minocycline-liposomes every five days were as effective in ameliorating clinical EAE as daily intraperitoneal injections of minocycline . Treatment of animals with PEG minocycline-liposomes significantly reduced the number of CNS-infiltrating leukocytes , and the overall expression of P14780 in the CNS . There was also a significant suppression of P14780 expression and proteolytic activity in splenocytes of treated animals , but not in CNS-infiltrating leukocytes . Thus , leukocytes gaining access to the brain and spinal cord require the same absolute amount of P14780 in all treatment groups , but minocycline decreases the absolute cell number . CONCLUSIONS : Our data indicate that less frequent injections of PEG minocycline-liposomes are an effective alternative pharmacotherapy to daily minocycline injections for the treatment of CNS autoimmune diseases . Also , inhibition of P14780 remains a promising treatment target in EAE and patients with MS . DB01017 exerts multiple inhibitory effects on vascular endothelial growth factor-induced smooth muscle cell migration : the role of P27361 /2 , PI3K , and matrix metalloproteinases . Widely used tetracycline antibiotics affect many cellular functions relevant to human vascular disease including cell proliferation , migration , and matrix remodeling . We examined whether minocycline inhibited human aortic smooth muscle cell ( HASMC ) migration induced by vascular endothelial growth factor ( P15692 ) . After the establishment of an optimal dose , minocycline treated HASMC were exposed to P15692 . HASMC migration , matrix metalloproteinase ( MMP ) -2 and P14780 activities , mitogen-activated protein kinase ( MAPK ) , and phosphatidylinositol 3-kinase ( PI3K ) phosphorylation were determined by smooth muscle cell ( SMC ) invasion assay , real-time polymerase chain reaction , zymograms , and Western blot analysis , respectively . We demonstrated that P15692 and platelet-derived growth factor ( PDGF ) -induced SMC migration in a dose-dependent manner . P14780 , but not P08253 , mRNA was increased during P15692 stimulation . P14780 activity was increased from 1.5- to 2.5-fold in a dose-dependent manner ( P < 0.05 ) . Both P27361 /2 and PI3K/AKt pathways were activated during P15692 -induced HASMCs migration . We then demonstrated that minocycline can inhibit P15692 -induced HASMC migration ( P < 0.05 ) . The effects may be through the inhibition of P14780 mRNA transcription , protein activities and downregulation of P27361 /2 and PI3K/Akt pathway phosphorylation . Our results indicated that minocycline exerts multiple effects on P15692 -induced SMC migration , including inhibition of P14780 mRNA transcription and protein activities and downregulating P27361 /2 and PI3K signal pathways , suggesting minocycline may be a potentially therapeutic approach to inhibit disease process induced angiogenesis . Protection of minocycline on early brain injury after subarachnoid hemorrhage in rats . DB01017 has been shown to be neuroprotective in cerebral ischemia and in other models of brain injury . Our goal is to observe the protection of minocycline on EBI after Q53FZ2 and the mechanism . 48 adult male SD rats were randomly divided into four groups : the sham-operated group , Q53FZ2 group , vehicle group ( Q53FZ2 +normal sodium ) , and minocycline group ( Q53FZ2 +minocycline ) . The Q53FZ2 model was induced by injecting 300 μl of autologous arterial blood into the prechiasmatic cistern . Expressions of P14780 in the hippocampus were examined at 24 h by western blot and zymography . Western blot and zymography showed that the expression of total and active P14780 increased dramatically at 24 h after Q53FZ2 compared with that of the sham group ( P < 0.01 ) . The clinical assessments got a lower score than that of the sham-operated group . After treated with minocycline , the expression of P14780 decreased significantly ( P < 0.01 vs. vehicle group ) , and the clinical assessments improved . We conclude that minocycline can protect EBI after Q53FZ2 , which may be related to the mechanism of inhibiting the expression of P14780 in the hippocampus . Antitumor activity of NPB001-05 , an orally active inhibitor of Bcr-Abl tyrosine kinase . Scientists are constantly searching for phytochemical compounds with anti-cancer activity . In this study , activity of plant extract NPB001-05 from Piper betle was tested on human chronic myelogenous leukemia ( CML ) xenograft models . NPB001-05 was active when dosed orally ( 500 mg/kg ) once or twice a day in xenograft tumor models . NPB001-05 showed activity to T315I tumor xenograft , where imatinib failed to show antitumor activity . NPB001-05 showed no relevant toxicity in animal models during 2 weeks exposure to drug . Responsive tumor showed inhibition of tyrosine kinase activity with lowered Bcr-Abl protein levels and increased apoptosis . Microarray based transcription profiling studies demonstrated that both imatinib and NPB001-05 dysregulated imatinib- responsive genes . NPB001-05 showed additional genes selectively dysregulated from ER stress , PI3K/AKT , MAPK pathways . Additionally , we tested gene expression of PI3K , P31749 , P05412 , P42574 and P35638 in K562 , BaF3P210( P11274 - P00519 ) and BaF3 P210( P11274 -ABLT315I) cell line treated for 6- and 12- hours with NPB001-05 and imatinib . The data indicates that NPB001-05 mediated cell death in K562 affects the function of ER stress . NPB001-05 shows antitumor activity with favorable toxicity profile . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . P01308 -induced NADPH oxidase activation promotes proliferation and matrix metalloproteinase activation in monocytes/macrophages . P01308 stimulates superoxide ( O(2)(-) ) production in monocytes and macrophages . However , the mechanisms through which insulin induces O(2)(-) production are not completely understood . In this study , we ( a ) characterized the enzyme and the pathways involved in insulin-stimulated O(2)(-) production in human monocytes and murine macrophages , and ( b ) analyzed the consequences of insulin-stimulated O(2)(-) production on the cellular phenotype in these cells . We showed that insulin stimulated O(2)(-) production , and promoted p47(phox) translocation to the plasma membrane . P01308 -induced O(2)(-) production and p47(phox) translocation were prevented in the presence of specific inhibitors of PI3K and PKC . P01308 -mediated NADPH oxidase activation stimulated P14780 activation in monocytes and cell proliferation in macrophages . The effect of insulin on these phenotypic responses was mediated through NFkappaB , p38MAPK , and P29323 1/2 activation . Small-interfering RNA-specific gene silencing targeted specifically against Nox2 reduced the cognate protein expression , decreased insulin-induced O(2)(-) production , inhibited the turn on of NFkappaB , p38MAPK , and P29323 1/2 , and reduced cell proliferation in macrophages . These findings suggest a pivotal role for NADPH oxidase in insulin-induced proliferation and proteolytic activation in monocytes and macrophages , respectively , and identify a pathway that may play a pathological role in hyperinsulinemic states .

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[ Asthma and aspirin ] . DB00945 ( ASA ) is an important cause of asthma so that ASA induced asthma ( AIA ) is considered a disease . Its prevalence is of 0.3-0.6 in the general population but it raises to 21 % in the asthmatic one . Middle aged female are the most affected . AIA generally begins with a non allergic rhinitis , complicated sometimes with polyps , that evolves secondarily in asthma . The disease is often so severe to need oral corticosteroids to be controlled . It persists independently to the intake of ASA . From a pathogenetic point of view the interaction of ASA with the arachidonic acid metabolism seems to be important . The inhibition of cyclo-oxygenase ( P36551 ) induces an activation of lypo-oxygenase with an increased synthesis of leukotrienes . In ASA intolerant patients there is also an activation of LTC4 synthetase , enzyme responsible for the synthesis of leukotrienes . Clinical history is very important to diagnose the disease but to confirm the diagnosis sometimes the provocation test is mandatory . Oral and bronchial challenges can be dangerous , while nasal challenge is safer even if must be better standardized . Patients must not use antiinflammatory drugs with the same mechanism of action of ASA ; P35354 inhibitors are generally well tolerated . Antileukotrienes are useful to treat asthma , in association with steroids . Desensitization can be used in very selected patients . DB00945 -exacerbated respiratory disease : evaluation and management . The clinical syndrome of aspirin-exacerbated respiratory disease ( AERD ) is a condition where inhibition of cyclooxygenase-1 ( P23219 ) induces attacks of upper and lower airway reactions , including rhinorrhea and varying degrees of bronchospasm and laryngospasm . Although the reaction is not IgE-mediated , patients can also present with anaphylactic hypersensitivity reactions , including hypotension , after exposure to P23219 inhibiting drugs . All patients with AERD have underlying nasal polyps and intractable sinus disease which may be difficult to treat with standard medical and surgical interventions . This review article focuses on the management of AERD patients with a particular emphasis on aspirin desensitization and continuous treatment with aspirin . Dual P36551 inhibition and upper gastrointestinal damage . DB00945 and non-aspirin NSAIDs injure the gastrointestinal tract principally as a result of their inhibition of prostaglandin synthesis . This is mediated via abrogation of the secretion of mucus and bicarbonate and by reduction in mucosal blood flow . Topical injury and inhibition of platelet thromboxane may also contribute respectively to damage and ulcer bleeding . Recognition of a second cyclooxygenase , P35354 , enabled drugs to be developed that selectively target this enzyme which is expressed in inflamed joints . These have proved to be effective treatments whilst causing little or no acute gastroduodenal injury and reduced ulcers and their complications . Future strategies may capitalise upon the phenomenon of substrate diversion of lipoxygenase products . Balanced cyclooxygenase/lipoxygenase inhibition maybe less harmful than cyclooxygenase inhibition . Also , nitric oxide can subserve many of the protective effects of prostaglandins and NO-donating NSAIDs are under evaluation . Increased platelet sensitivity among individuals with aspirin resistance - platelet aggregation to submaximal concentration of arachidonic acid predicts response to antiplatelet therapy . DB00945 ' resistance ' ( AR ) is a phenomenon of uncertain etiology describing decreased platelet inhibition by aspirin . We studied whether ( i ) platelets in AR demonstrate increased basal sensitivity to a lower degree of stimulation and ( ii ) platelet aggregation with submaximal stimulation could predict responses to aspirin . Serum thromboxane B(2) ( TxB(2) ) levels and platelet aggregation with light transmission aggregometry ( P01374 ) were measured at baseline and 24 hours after 325 mg aspirin administration in 58 healthy subjects . AR was defined as the upper sixth of P01374 ( > or = 12 % ) to 1.5 mM AA . Baseline platelet aggregation with submaximal concentrations of agonists [ ADP 2 microM , arachidonic acid ( AA ) 0.75 mM , collagen 0.375 and 0.5 microg/ml ] was greater in AR subjects compared with non-AR subjects , but not with higher concentrations ( ADP 5 microM and 20 microM , AA 1.5 mM and collagen 1 microg/ml ) . Post-aspirin platelet aggregation was elevated in AR subjects with both submaximal and maximal stimulation . Baseline and post-aspirin serum TxB(2) were higher in AR subjects and decreased further with ex-vivo P23219 inhibition , suggesting incompletely suppressed P23219 activity . Pre-aspirin platelet aggregation to 0.75 AA demonstrated a dichotomous response with 29/58 subjects having aggregation < or = 15 % and 29/58 subjects having aggregation > or = 75 % . In the high aggregation group 28 % had AR compared to 6 % in the non-AR group ( p = 0.04 ) . In conclusion , platelets in AR subjects demonstrate increased basal sensitivity to submaximal stimulation , which could predict responses to antiplatelet therapy . Effect of non-selective , non-steroidal anti-inflammatory drugs and cyclo-oxygenase-2 selective inhibitors on the PFA-100 closure time . The place of cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drugs ( NSAIDs ) in the peri-operative period remains under discussion . Due to the absence of P35354 in platelets , the risk of bleeding in patients who use selective NSAIDs is thought to be decreased . We studied the influence of aspirin , diclofenac , lornoxicam and rofecoxib on the in vitro bleeding time using the platelet function analyser ( PFA-100 ) . The PFA-100 simulates the process of platelet adhesion and aggregation after vascular injury in vitro . Measurements in 43 volunteers were performed at three time points : before , 3 h , and 12 h after oral ingestion of one of the randomly assigned study medications . DB00945 , diclofenac and lornoxicam had a significant effect on the in vitro closure time , while rofecoxib did not show this effect . This supports the use of P35354 selective drugs in the peri-operative period to minimise the risk of bleeding . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . In vivo effects of meloxicam and aspirin on blood , gastric mucosal , and synovial fluid prostanoid synthesis in dogs . OBJECTIVE : To evaluate in vivo activity in dogs of meloxicam or aspirin , previously shown in vitro to be a selective cyclooxygenase-2 ( P35354 ) inhibitor ( P23219 sparing drug ) , or a nonselective P36551 inhibitor , respectively . ANIMALS : 12 male dogs with unilateral osteoarthritis of the stifle joint . PROCEDURE : Each dog was treated in a crossover design with aspirin or meloxicam for 21 days . DB00917 ( DB00917 ) concentrations were measured at days 0 ( baseline ) , 7 , and 21 of each treatment period in lipopolysaccharide ( LPS ) -stimulated blood , synovial fluid collected by arthrocentesis , and endoscopic gastric mucosal biopsy specimens . Thromboxane B2 ( TXB2 ) was evaluated in blood on days 0 , 7 , and 21 of each treatment period . RESULTS : DB00945 administration significantly suppressed DB00917 concentrations in blood , gastric mucosa , synovial fluid , and suppressed TXB2 concentration in blood at days 7 and 21 . Meloxicam administration significantly suppressed DB00917 concentrations in blood and synovial fluid at days 7 and 21 , but had no effect on concentrations of TXB2 in blood or DB00917 in gastric mucosa . Suppression of LPS-stimulated DB00917 concentrations in blood and synovial fluid by aspirin and meloxicam administration is consistent with activity against the P35354 isoenzyme . Suppression of concentrations of DB00917 in the gastric mucosa and TXB2 in blood by aspirin administration is consistent with activity against P23219 . Meloxicam , in contrast , had a minimal effect on functions mediated by P23219 . CONCLUSIONS AND CLINICAL RELEVANCE : Meloxicam acts in vivo in dogs as a P23219 sparing drug on target tissues by sparing gastric DB00917 synthesis while retaining antiprostaglandin effects within inflamed joints . DB00945 -triggered Lipoxin A4 attenuates mechanical allodynia in association with inhibiting spinal O60674 / P40763 signaling in neuropathic pain in rats . DB00945 -triggered Lipoxin A4 ( ATL ) , as a Lipoxin A4 ( LXA4 ) epimer , is endogenously produced by aspirin-acetylated cycloxygenase-2 ( P35354 ) and plays a vital role in endogenous anti-inflammation via the P25090 ( Q96JZ2 ) . Recent investigations have indicated that spinal neuroinflammation and the activation of the Janus Kinase 2 ( O60674 ) /Signal Transducers and Transcription Activators 3 ( P40763 ) signaling pathway are involved in neuropathic pain states . However , the effect of ATL on neuroinflammation and O60674 / P40763 signaling in chronic constriction injury ( CCI ) -induced neuropathic pain in rats has not been well-studied . The present study demonstrated the anti-inflammatory and analgesic effect of ATL on neuropathic pain and assessed the role of spinal O60674 / P40763 signaling on the effect of ATL . Intrathecal administration of ATL significantly attenuated mechanical allodynia via spinal Q96JZ2 and inhibited the upregulation of pro-inflammatory cytokines ( IL-1β , P05231 , and P01375 -α ) on day 7 of CCI surgery . In addition , ATL markedly suppressed the upregulation of p- P40763 induced by the neuropathic pain . Blockade of O60674 - P40763 signaling with intrathecal administration of the O60674 inhibitor AG490 or the P40763 inhibitor S3I-201 clearly reduced mechanical allodynia and the upregulation of pro-inflammatory cytokines in CCI rats . Interestingly , inhibition of O60674 / P40763 signaling via ATL or the specific signaling inhibitor ( AG49 , S3I-201 ) further promoted the increased expression of suppressor of cytokine signaling 3 ( O14543 ) mRNA in the spinal cord induced by CCI surgery . Taken together , our results suggested that the analgesic effect of ATL was mediated by inhibiting spinal O60674 / P40763 signaling and hence the spinal neuroinflammation in CCI rats . Suppression of inducible cyclooxygenase 2 gene transcription by aspirin and sodium salicylate . The pharmacological action of salicylate can not be explained by its inhibition of cyclooxygenase ( P36551 ) activity . In this report , the effects of aspirin and sodium salicylate on P35354 expressions in human umbilical vein endothelial cells and foreskin fibroblasts were evaluated . DB00945 and sodium salicylate at therapeutic concentrations equipotently blocked P35354 mRNA and protein levels induced by interleukin-1beta and phorbol 12-myristate 13-acetate . The suppressing effect was more pronounced in cultured cells deprived of fetal bovine serum for 24 h , suggesting that it may be cell cycle related . Salicylate inhibited nascent P35354 transcript synthesis but had no effect on P35354 mRNA stability . It inhibited P35354 promoter activity in a concentration-dependent manner . In mice pretreated with aspirin ( 10 and 30 mg/kg ) , followed by challenge with lipopolysaccharide , P35354 mRNA expression in peritoneal macrophages was markedly suppressed . These findings suggest that salicylate exerts its antiinflammatory action in part by suppressing P35354 induction , thereby reducing the synthesis of proinflammatory prostaglandins . Limitations of current therapies to prevent thrombosis : a need for novel strategies . Bleeding limits the benefit of current anti-platelet drugs for preventing heart attacks and stroke . DB00945 and clopidogrel , the two most widely prescribed anti-platelet drugs , are metabolized to active compounds that covalently and irreversibly modify their respective therapeutic targets ( P23219 and Q9H244 ) . The enduring effects of aspirin and clopidogrel are of concern in patients receiving anti-platelet therapy who require emergency surgery as this places them at greater risk of haemorrhage . As clopidogrel must be activated by cytochrome P450 metabolism , recent pharmacogenomic studies have revealed that patients lacking a functional allele of P33261 derive no therapeutic benefit from the drug . Prasugrel , a second generation thienopyridine , whose bioconversion is not affected by CYP genetic polymorphism , demonstrates improved clinical benefit , but with increased bleeding risk . Anti-platelet drugs currently in cardiovascular trials that may have reduced bleeding risk include reversible Q9H244 antagonists ( cangrelor , ticagrelor , and elinogrel ) , a PAR1 antagonist ( P35240 530 348 ) and an EP3 antagonist ( DG-041 ) . The platelet EP3 receptor for prostaglandin E(2) is an attractive therapeutic target as EP3 antagonists may selectively avert thrombosis over atherosclerotic plaques without affecting bleeding risk . Biochemical pharmacology of nonsteroidal anti-inflammatory drugs . DB00945 and conventional nonsteroidal anti-inflammatory drugs are nonselective inhibitors of cyclooxygenase-1 ( P23219 ) and P35354 enzymes . Two classes of selective P35354 inhibitors : ( 1 ) sulfonamides , such as L-745,337 , and ( 2 ) tricyclic methyl sulfone derivatives , such as SC58125 , have been developed . X-ray crystal structures of P23219 and P35354 have provided valuable information regarding the structural basis for their P35354 selectivity . These compounds have less gastrointestinal complications in animal experiments . Their clinical efficacy and side-effects are being evaluated . Salicylate has very weak activity against either P36551 isoform and yet possesses anti-inflammatory actions . Recent studies indicate that it suppresses the expression of genes involved in inflammation . These activities may provide a plausible explanation for the pharmacological dilemma and , furthermore , may represent novel mechanisms for controlling inflammation . Inhibition of P08908 receptor-dependent cell survival by DB02527 /protein kinase A : role of protein phosphatase 2A and Bax . Serotonergic 5-HT(1A) receptor signaling leading to nuclear factor-kappaB ( NF-kappaB ) activation appears to be critical for cell survival . Adenylyl cyclase and protein kinase A ( AC/PKA ) are effectors of the 5-HT(1A) receptor that are inhibited by Galpha(i) subunits . Conversely , Gbetagamma(i) subunits downstream from the 5-HT(1A) receptor participate in the activation of extracellular signal-regulated kinases ( P27361 /2 ) , phosphatidylinositol 3-kinase ( PI3K ) , Akt , and NF-kappaB . To model the contribution of pro- and antiapoptotic signaling cascades downstream of activated 5-HT(1A) receptor in cell survival , Chinese hamster ovarian ( CHO ) cells were employed that exogenously overexpress 5-HT(1A) receptors . Stimulation with the 5-HT(1A) receptor agonist 8-OH-DPAT and pharmacological agonists of AC induced PKA and protein phosphatase 2A ( PP2A ) activity , which in turn inhibited : Akt activity , P25963 degradation , nuclear translocation of NF-kappaB , and expression of P98170 ( P98170 / P98170 ) . Pharmacological inhibition of PP2A with calyculin A potentiated Akt activity while attenuating P27361 /2 signaling via increased inhibitory phosphorylation of Raf ( pSer259 ) . In contrast , increased DB02527 levels enhanced Bax translocation to the mitochondria , resulting in the release of cytochrome c , caspase-3 activation , and apoptosis induction . Our data suggest a central role of DB02527 /PKA-dependent PP2A in shifting the homeostasis of intracellular signaling downstream of activated 5-HT(1A) receptor toward cell death in biological systems linked to neuropsychiatric disorders . P35354 in newborn hyperoxic lung injury . Supraphysiological O2 concentrations , mechanical ventilation , and inflammation significantly contribute to the development of bronchopulmonary dysplasia (BPD).Exposure of newborn mice to hyperoxia causes inflammation and impaired alveolarization similar to that seen in infants with BPD.Previously , we demonstrated that pulmonary cyclooxygenase-2 ( P35354 ) protein expression is increased in hyperoxia-exposed newborn mice.The present studies were designed to define the role of P35354 in newborn hyperoxic lung injury.We tested the hypothesis that attenuation of P35354 activity would reduce hyperoxia-induced inflammation and improve alveolarization.Newborn C3H/HeN micewere injected daily with vehicle , aspirin ( nonselective P35354 inhibitor ) , or celecoxib ( selective P35354 inhibitor ) for the first 7 days of life.Additional studies utilized wild-type ( C57Bl/6 , P35354 (+/+) ) , heterozygous ( P35354 (+/-) ) , and homozygous ( P35354 (-/-) ) transgenic mice.Micewere exposed to room air ( 21 % O2 ) or hyperoxia ( 85 % O2 ) for 14 days. DB00945 -injected and P35354 (-/-) pups had reduced levels of monocyte chemoattractant protein ( P13500 ) in bronchoalveolar lavage fluid (BAL).Both aspirin and celecoxib treatment reduced macrophage numbers in the alveolar walls and air spaces. DB00945 and celecoxib treatment attenuated hyperoxia-induced P36551 activity , including altered levels of prostaglandin (PG)D2 metabolites.Decreased P36551 activity , however , did not prevent hyperoxia-induced lung developmental deficits.Our data suggest thatincreased P35354 activity may contribute to proinflammatory responses , including macrophage chemotaxis , during exposure to hyperoxia.Modulation of P35354 activity may be a useful therapeutic target to limit hyperoxia-induced inflammation in preterm infants at risk of developing BPD . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . Nonsteroidal anti-inflammatory drugs : adverse effects and their prevention . OBJECTIVES : To discuss nonsteroidal anti-inflammatory drugs ( NSAIDs ) , their history , development , mode of action , toxicities , strategies for the prevention of toxicity , and future developments . METHODS : Medline search for articles published up to 2007 , using the keywords acetylsalicylic acid , aspirin , NSAIDs , cyclooxygenase 2 , adverse effects , ulcer , and cardiovascular . RESULTS : NSAIDs are 1 of the oldest , most successful drugs known to modern medicine . They are effective for alleviating pain , fever , and inflammation by inhibiting prostaglandin synthesis . DB00945 , by its irreversible inhibition of blood platelet function , is also effective in the prevention of cardiovascular disease . NSAIDs may cause gastrointestinal ulcers , serious cardiovascular events , hypertension , acute renal failure , and worsening of preexisting heart failure . These adverse effects may be prevented by limiting NSAID dosage and duration and by performing individual risk assessments and treating patients accordingly . Those at risk for gastroduodenal ulcers may be treated with concomitant proton-pump inhibitors , misoprostol and/or P35354 selective NSAIDs . Those at risk for cardiovascular events may be treated with naproxen and a proton-pump inhibitor or misoprostol , but should best avoid NSAID use altogether . CONCLUSIONS : Physicians should always prescribe the lowest effective dose for the shortest possible time and must take into account both the gastrointestinal and the cardiovascular risks of individual patients when prescribing NSAIDs . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Cyclooxygenase isozymes are expressed in human myeloma cells but not involved in anti-proliferative effect of cyclooxygenase inhibitors . Considering possible tumorigenic activity of cyclooxygenase ( P36551 ) isozymes in myeloma , we examined expression levels of P23219 and -2 in seven human myeloma cell lines ( Q5SW96 -77 , IM-9 , RPMI-8226 , HPC , HS-Sultan , TSPC-1 , and U-266 ) . As analyzed by reverse transcriptase-polymerase chain reaction ( RT-PCR ) , all the cell lines constitutively expressed P23219 , while P35354 levels markedly varied among different cell lines . Induction of P35354 by phorbol ester was observed in RPMI-8226 and HPC cells . In contrast , P35354 was constitutively expressed in Q5SW96 -77 and IM-9 cells . Moreover , the high expression level of P35354 protein in Q5SW96 -77 cells was verified by Western blotting . Intact cells of Q5SW96 -77 converted 14C-labeled arachidonic acid to prostaglandin E2 , F2alpha , and D2 , and this activity was dose-dependently inhibited by selective P35354 inhibitors ( SC-58125 and NS-398 ) , a non-selective P36551 inhibitor ( indomethacin ) , and relatively high concentrations of a selective P23219 inhibitor ( SC-560 ) . These P36551 inhibitors also suppressed the proliferation of Q5SW96 -77 cells , but significant suppression was seen only at 100 microM , a much higher concentration than those sufficient for the P36551 inhibition . Moreover , proliferation of the myeloma cells lacking P35354 was also suppressed by 100 microM of SC-58125 . These results suggested that the anti-proliferative effect of the P36551 inhibitors is independent of the inhibition of P35354 . DB00945 -triggered 15-epi-lipoxin A4 predicts cyclooxygenase-2 in the lungs of LPS-treated mice but not in the circulation : implications for a clinical test . Inhibition of cyclooxygenase ( P36551 ) -2 increases cardiovascular deaths . Identifying a biomarker of P35354 is desirable but difficult , since P23219 and P35354 ordinarily catalyze formation of an identical product , prostaglandin H2 . When acetylated by aspirin , however , P35354 ( but not P23219 ) can form 15(R)-HETE , which is metabolized to aspirin-triggered lipoxin ( ATL ) , 15-epi-lipoxin A4 . Here we have used P23219 - and P35354 -knockout mice to establish whether plasma ATL could be used as a biomarker of vascular P35354 in vivo . Vascular P35354 was low but increased by LPS ( 10 mg/kg ; i.p ) . DB00945 ( 10 mg/kg ; i.v. ) inhibited P23219 , measured as blood thromboxane and P35354 , measured as lung DB00917 . DB00945 also increased the levels of ATL in the lungs of LPS-treated wild-type C57Bl6 mice ( vehicle : 25.5±9.3 ng/ml ; 100 mg/kg : 112.0±7.4 ng/ml ; P < 0.05 ) . Despite this , ATL was unchanged in plasma after LPS and aspirin . This was true in wild-type as well as P23219 (-/-) and P35354 (-/-) mice . Thus , in mice in which P35354 has been induced by LPS treatment , aspirin triggers detectable 15-epi-lipoxin A4 in lung tissue , but not in plasma . This important study is the first to demonstrate that while ATL can be measured in tissue , plasma ATL is not a biomarker of vascular P35354 expression . DB00945 and P35354 inhibitor nimesulide potentiate adrenergic contractions of human gastroepiploic artery . BACKGROUND : The aim of the present study was to evaluate the intervention of P23219 - and P35354 -derived prostaglandins in the responses of human gastroepiploic artery to sympathetic stimulation and norepinephrine . METHODS : Rings of human gastroepiploic artery were obtained from 45 patients ( 26 men and 19 women ) undergoing gastrectomy . The rings were suspended in organ baths for isometric recording of tension . We studied the responses to electrical field stimulation , norepinephrine , and acetylcholine , in the absence and presence of P23219 or P35354 inhibition . RESULTS : The P23219 and P35354 inhibitor aspirin at high concentrations ( 10(-6) to 10(-5) mol/L ) and the P35354 inhibitor nimesulide ( 10(-6) mol/L ) potentiated the contractile responses of the arterial rings to sympathetic neurogenic stimulation and norepinephrine . In contrast , lower concentrations of aspirin ( 10(-8) to 10(-7) mol/L ) or the P23219 inhibitor SC-560 ( 3 x10(-8) mol/L ) did not affect these responses . The vascular relaxation induced by acetylcholine was not affected by P23219 and P35354 inhibition . CONCLUSIONS : The results provide functional evidence that vasodilator prostaglandins are active components of the response of human gastroepiploic artery to neurogenic stimulation and norepinephrine . DB00945 at high concentrations and the P35354 selective inhibitor nimesulide potentiated the contractile response of gastroepiploic artery to adrenergic stimulation by inhibiting P35354 -derived P06744 (2) . DB00945 at low concentrations and the P23219 selective inhibitor SC-560 did not modify the contractile responses , possibly due to minor importance of vasoconstrictor prostaglandins ( TXA(2) ) as active components of the response of gastroepiploic artery to adrenergic stimulation . P02751 -induced P35354 mediates P08253 expression and invasiveness of rhabdomyosarcoma . Although accumulating evidence suggests the importance of cyclooxygenase-2 ( P35354 ) and prostaglandin E(2) ( PGE(2) ) in the pathogenesis of many cancers , the mechanism by which this enzyme and its metabolite promote cancer progression is unknown . In this study , we investigated the role of P35354 in fibronectin-induced up-regulation of rhabdomyosarcoma matrix metalloproteinase ( MMP ) -2 activity and cellular invasiveness . We tested three human rhabdomyosarcoma cell lines : RMS559 , RD , and SJRH30 . Cell attachment to fibronectin up-regulated both P35354 expression and PGE(2) production and concomitantly enhanced P08253 activity . Exogenous PGE(2) stimulated P08253 promoter activity , increased P08253 expression , and increased cellular invasiveness . DB00945 and rofecoxib ( non-selective and selective P35354 inhibitor , respectively ) each abolished fibronectin-associated induction of P08253 and induced dose-dependent reductions in cellular invasiveness . These data implicated a role for inducible P35354 and PGE(2) in the regulation of rhabdomyosarcoma cellular invasiveness and P08253 activity . Single-walled carbon nanotubes ( SWCNTs ) enhance DB00761 - , acetylcholine- , and serotonin-induced contractions and evoke oxidative stress on rabbit ileum . We examined the effects of intravenous administration of purified arc-discharge single-walled carbon nanotubes ( SWCNTs ) on rabbit ileum to establish the possibility of using these SWCNTs as cell markers or drug carriers for the treatment of intestinal diseases . The SWCNT purification process eliminated carbonaceous impurities and decreased the amount of metals . SWCNTs increased the contractile responses induced by DB00761 , acetylcholine ( ACh ) , and serotonin ( 5-HT ) in rabbit ileum . Verapamil , apamin , glibenclamide , quinine and charybdotoxin reduced the contractile responses induced by ACh and 5-HT in ileum from rabbits treated with SWCNTs , indicating that voltage-dependent Ca2+ channels and small , intermediate , and large-conductance Ca(2+)-activated , DB00171 -sensitive , and voltage-dependent K+ channels are involved in these effects . Atropine and hexamethonium reduced the ACh response , indicating that muscarinic and nicotinic receptors are involved in this effect . DB00904 and GR 113808 reduced the 5-HT response , indicating that serotonin 5- Q9H205 and Q13639 receptors are involved in this effect . SWCNTs increased the malondialdehyde plus 4-hydroxyalkenals and carbonyl levels in rabbit plasma and ileum , indicating that SWCNTs produce oxidative stress . SWCNTs did not produce relevant histological changes or modify the levels of the inflammatory mediators P35228 and P35354 in the ileum . In conclusion , this study demonstrates that the intravenous administration of SWCNTs can evoke oxidative stress and affect contractility in rabbit ileum . These effects could reduce the possibility of using the arc-discharge SWCNTs as cell markers or drug carriers to treat intestinal diseases . Role of the JAK- P35610 pathway in protection against myocardial ischemia/reperfusion injury . The Janus kinase ( JAK ) -signal transducers and activators of transcription ( P35610 ) pathway is a stress-responsive mechanism that transduces signals from the cell surface to the nucleus , thereby modulating gene expression . Recent studies have demonstrated that myocardial ischemia and reperfusion induce rapid activation of this pathway . Although the functional consequences of this event remain to be elucidated , there is emerging evidence that JAK- P35610 signaling plays an important role in the development of the cardioprotected phenotype associated with ischemic preconditioning . Specifically , brief episodes of myocardial ischemia/reperfusion activate P23458 and O60674 , followed by recruitment of P42224 and P40763 , resulting in transcriptional upregulation of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) , which then mediate the infarct-sparing effects of the late phase of preconditioning . The present review focuses on this novel cardioprotective role of JAK- P35610 signaling and on its potential exploitation for developing therapeutic strategies aimed at limiting ischemia/reperfusion injury . [ Cyclooxygenase ( P36551 ) -2 selective inhibitors : aspirin , a dual P23219 / P35354 inhibitor , to P35354 selective inhibitors ] . DB00945 was developed as a non-steroidal anti-inflammatory drug ( NSAID ) in 1899 . During the century after that , aspirin has been found to show its anti-inflammatory , analgesic and anti-pyretic activities by reducing prostaglandins biosynthesis through inhibition of cyclooxygenase ( P36551 ) ; and then P36551 was found to be constituted of two isoforms , constitutive P23219 and inducible P35354 . Currently , novel NSAIDs , acting through selective inhibition of P35354 , that have efficacy as excellent as aspirin with significantly lower incidence of gastrointestinal adverse effects are available in America and some other countries , but not in Japan . Physiological and pathophysiological roles of P23219 and P35354 have been explained from studies in experimental animals , but there are many differences in species and diseases between animals and humans . Thus , physiological and pathophysiological roles of P35354 were considered from the standpoint of clinical effects of the two latest P35354 selective inhibitors , celecoxib and rofecoxib , on inflammation , pain , fever and colorectal cancer together with their adverse effects on gastrointestinal , renal and platelet functions ; and the usefulness and limits of P35354 -selective inhibitors were discussed with the trends of new NSAIDs development . Mode of action of aspirin as a chemopreventive agent . DB00945 taken for several years at doses of at least 75 mg daily reduced long-term incidence and mortality due to colorectal cancer . The finding of aspirin benefit at low-doses given once daily , used for cardioprevention , locates the antiplatelet effect of aspirin at the center of its antitumor efficacy . In fact , at low-doses , aspirin acts mainly by an irreversible inactivation of platelet cyclooxygenase ( P36551 ) -1 in the presystemic circulation , which translates into a long-lasting inhibition of platelet function . Given the short half-life of aspirin in the human circulation ( approximately 20 min ) and the capacity of nucleated cells to resynthesize the acetylated P36551 -isozyme(s) , it seems unlikely that a nucleated cell could be the target of aspirin chemoprevention . These findings convincingly suggest that colorectal cancer and atherothrombosis may share a common mechanism of disease , i.e. platelet activation in response to epithelial ( in tumorigenesis ) and endothelial ( in tumorigenesis and atherothrombosis ) injury . Activated platelets may also enhance the metastatic potential of cancer cells ( through a direct interaction and/or the release of soluble mediators or exosomes ) at least in part by inducing the overexpression of P35354 . P36551 -independent mechanisms of aspirin , such as the inhibition of NF-kB signaling and Wnt/β-catenin signaling and the acetylation of extra- P36551 proteins , have been suggested to play a role in its chemopreventive effects . However , their relevance remains to be demonstrated in vivo at clinical doses . Acetylation of prostaglandin H2 synthases by aspirin is inhibited by redox cycling of the peroxidase . DB00945 exerts its unique pharmacological effects by irreversibly acetylating a serine residue in the cyclooxygenase site of prostaglandin-H2-synthases ( PGHSs ) . Despite the irreversibility of the inhibition , the potency of aspirin varies remarkably between cell types , suggesting that molecular determinants could contribute to cellular selectivity . Using purified enzymes , we found no evidence that aspirin is selective for either of the two PGHS isoforms , and we showed that hydroperoxide substrates of the PGHS peroxidase inhibited the rate of acetylation of P23219 by 68 % . Using P23219 reconstituted with cobalt protoporphyrin , a heme devoid of peroxidase activity , we demonstrated that reversal by hydroperoxides of the aspirin-mediated acetylation depends upon the catalytic activity of the PGHS peroxidase . We demonstrated that inhibition of P35354 by aspirin in cells in culture is reversed by 12-hydroperoxyeicosatetraenoic acid dose-dependently ( ED50=0.58+/-0.15 microM ) and that in cells with high levels of hydroperoxy-fatty acids ( RAW264.7 ) the efficacy of aspirin is markedly decreased as compared to cells with low levels of hydroperoxides ( A549 ; IC50s=256+/-22 microM and 11.0+/-0.9 microM , respectively ) . Together , these findings indicate that acetylation of the PGHSs by aspirin is regulated by the catalytic activity of the peroxidase , which yields a higher oxidative state of the enzyme . Selective inhibitors of Q02750 /ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells . The nonsteroidal anti-inflammatory drug ( NSAID ) sulindac prevents experimental colon cancer and can regress precancerous polyps in humans . Sulindac sulfide inhibits cyclooxygenase ( P36551 ) -mediated prostaglandin synthesis and retards the growth of cultured colon cell lines primarily by inducing apoptosis . Given the known role of mitogen-activated protein kinase ( MAPK ) in signal transduction and the regulation of cell survival and death , we determined the effect of sulindac sulfide on MAPK activation , P35354 expression , and apoptosis induction in HCA-7 human colon cancer cells . Sulindac sulfide treatment was associated with activation of ERKp44/42 and p38 MAPK in a dosage- and time-dependent manner , and also activated upstream MEK . Similar results were seen in HCT-15 cells and also with the selective P35354 inhibitor NS398 . ERKp44/42 and p38 activation were accompanied by an induction of P35354 protein expression . Selective inhibitors of sulindac sulfide-induced ERKp44/42 ( PD98059 ) and p38 MAPK ( SB203580 ) activation also suppressed the induction of P35354 by this NSAID . Furthermore , both MAPK inhibitors significantly augmented sulindac sulfide-induced apoptosis , as did suppression of constitutive P35354 using antisense oligonucleotides . In conclusion , MEK/ P29323 and p38 MAPK activation mediate P35354 induction by sulindac sulfide . Selective inhibitors of these MAPKs potentiate apoptosis induction by this NSAID , suggesting a novel strategy for the prevention or treatment of colorectal cancer . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Pathogenesis of aspirin-exacerbated respiratory disease . The underlying respiratory disease is activated by unknown mechanism and results in an intense infiltration of mast cells and eosinophils into the entire respiratory mucosa . These cells synthesize leukotrienes ( LTs ) at a very high rate and mast cells also release histamine and tryptase and synthesize P52209 (2) a vasodilator and bronchoconstrictor . Furthermore , AERD patients under synthesize from arachidonic acid ( AA ) a peculiar product called lipoxins , which opposes inflammation generated by leukotrienes . Finally , cysLT1 receptors are over expressed and highly responsive to LTE(4) , further augmenting the underlying inflammatory disease . This inflammatory condition is partly inhibited by synthesis of PGE(2) through P23219 . PGE(2) partially inhibits 5-lipogygenase conversion of AA to P01374 (4) and blocks release of histamine and tryptase from mast cells . When P36551 -l is inhibited by ASA or NSAIDs , PGE(2) synthesis stops and an enormous release of histamine and synthesis of LTs occurs . The upper respiratory reaction is mediated by both histamine and LTs but the bronchospastic reaction is mediated by LTs . The systemic effects of flush , gastric pain and hives are mediated by histamine . DB00945 desensitization can not be explained by disappearance of LT synthesis since urine LTE(4) levels are still elevated at acute ASA desensitization . However , mast cell products such as histamine , tryptase and P52209 (2) are no longer released or synthesized at acute desensitization . It is more likely that a diminution in number or function of cysLT receptors accounts for the diminished inflammatory response found in ASA desensitization . Bioassay-guided isolation of an alkaloid with antiangiogenic and antitumor activities from the extract of Fissistigma cavaleriei root . Fissistigma cavaleriei ( Levl ) Rehd ( Annonaceae ) is used as a folklore medicine for treatment of inflammation , arthritis , and tuberculosis by Miao people in China . In the present study , the antiangiogenic activity of F. cavaleriei was investigated . The chorioallantoic membrane of the fertilized hen 's egg ( P62158 assay ) was used to determine antiangiogenic activity of the plant extract . Compound ( 1 ) , a compound with antiangiogenic activity , was isolated by bioassay-guided fractionation from F. cavaleriei for the first time . The structure of compound ( 1 ) was elucidated on the basis of spectroscopic methods . Colorimetric P36551 ( ovine ) inhibitor screening assay was used to determine its inhibitory effect on P23219 and P35354 . MTT and Sulforhodamine B assays were used to investigate its cytotoxic effects on tumor cell lines . As a result , compound ( 1 ) showed a selectively inhibiting effect on P35354 and could inhibit the growth of tumor cells in vitro . The antitumor activity of compound ( 1 ) was further confirmed by the observation that compound ( 1 ) administration significantly inhibited the growth of S-180 cells in mice . Moreover , compound ( 1 ) was able to enhance the antitumor activity of doxorubicin in the mice bearing with S-180 cells while combined with doxorubicin . In conclusion , compound ( 1 ) is a multi-target molecule and further experimental investigations are needed to determine whether it can be used as a lead molecule for tumor treatment . Mechanisms of aspirin resistance . DB00945 is integral to the secondary prevention of cardiovascular disease and acts to impair the development of platelet-mediated atherothromboembolic events by irreversible inhibition of platelet cyclooxygenase-1 ( P23219 ) . Inhibition of this enzyme prevents the synthesis of the potent pro-aggregatory prostanoid thromboxane A2 . A large number of patients continue to experience atherothromboembolic events despite aspirin therapy , so-called ' aspirin treatment failure ' , and this is multifactorial in aetiology . Approximately 10 % however do not respond appropriately to aspirin in a phenomenon known as ' aspirin resistance ' , which is defined by various laboratory techniques . In this review we discuss the reasons for aspirin resistance in a systematic manner , starting from prescription of the drug and ending at the level of the platelet . Poor medication adherence has been shown to be a cause of apparent aspirin resistance , and may in fact be the largest contributory factor . Also important is high platelet turnover due to underlying inflammatory processes , such as atherosclerosis and its complications , leading to faster regeneration of platelets , and hence of P23219 , at a rate that diminishes the efficacy of once daily dosing . Recent developments include the identification of platelet glycoprotein IIIa as a potential biomarker ( as well as possible underlying mechanism ) for aspirin resistance and the discovery of an anion efflux pump that expels intracellular aspirin from platelets . The absolute as well as relative contributions of such factors to the phenomenon of aspirin resistance are the subject of continuing research . Possible mechanisms of drug-induced aspirin and clopidogrel resistance . DB00945 ( ASA ) and clopidogrel have been identified as standard of care in the prevention of major cardiovascular events . DB00945 irreversibly inhibits the cyclooxygenase-1 ( P23219 ) enzyme , whereas non-steroidal anti-inflammatory drugs ( NSAIDs ) reversibly inhibit the P23219 enzyme . An analysis of the literature revealed a statistically significant decrease in clinical benefit of ASA with concomitant administration of ibuprofen . Another NSAID , diclofenac , showed minimal effect on the inhibition of platelet aggregation when administered with ASA . Furthermore , the selective P35354 inhibitor , rofecoxib , was not shown to influence the effect of ASA . DB00758 is metabolized to an active thiol metabolite by the CYP 3A4 enzyme . Some HMG DB01992 reductase inhibitors have the ability to inhibit the CYP 3A4 enzyme , which can result in a possible interaction if administered concomitantly with clopidogrel . Studies have demonstrated clopidogrel 's platelet inhibition being significantly attenuated by atorvastatin . However in a post-hoc analysis , it was demonstrated that there was no difference in clinical outcomes between patients taking clopidogrel and P04035 inhibitors metabolized by and not metabolized by CYP 3A4 . Data suggest that the interaction observed involving clopidogrel and P04035 inhibitors appears to be significant in-vitro . Therefore , practitioners should advise patients receiving chronic aspirin therapy to limit the use of ibuprofen and may consider concomitant administration of clopidogrel with P04035 inhibitors without regard for the drug interaction . The intent of this paper is to review the literature discussing possible mechanisms of drug-induced aspirin and clopidogrel resistance and discuss whether the interactions translate into clinical effects . Anti-microinflammatory lipid signals generated from dietary N-3 fatty acids via cyclooxygenase-2 and transcellular processing : a novel mechanism for NSAID and N-3 PUFA therapeutic actions . DB00945 therapy inhibits prostaglandin biosynthesis ; yet via acetylation of cyclooxygenase 2 ( P35354 ) it leads to bioactive lipoxins epimeric at carbon 15 ( 15-epi-LX , also termed aspirin-triggered lipoxin or ATL ) . Here , we review our findings indicating that inflammatory exudates from mice treated with omega-3 PUFA and aspirin ( ASA ) generate a novel array of bioactive lipid signals . Also , human endothelial cells , both HUVEC and microvascular , with upregulated P35354 and treated with ASA converted C20:5 omega-3 to 18R-hydroxyeicosapentaenoic acid ( HEPE ) and 15R-HEPE . Human PMN activated with serum treated zymosan ( Q11206 ) utilized each of these R-HEPEs to generate novel classes of trihydroxy-containing mediators including 5-series 15R-LX and 5,12,18R-triHEPE . The novel products were potent inhibitors of human PMN transendothelial migration and infiltration of PMN in dorsal air pouches in vivo . In addition to ASA , both acetaminophen and indomethacin also permitted 18R-HEPE and 15R-HEPE generation with recombinant human P35354 as well as omega-5 and omega-9 oxygenations of other fatty acids that act on leukocytes , platelets and endothelial cells . These findings establish new transcellular routes for producing arrays of lipid mediators via P35354 -NSAIDs and cell-cell interactions that impact microinflammation . Moreover , they provide novel mechanism(s) that could underlie the many reported therapeutic benefits of omega-3 dietary supplementation of interest in inflammation , cancer , and vascular disorders . The Multifaceted Clinical Readouts of Platelet Inhibition by Low-Dose DB00945 . Inactivation of platelet cyclooxygenase ( P36551 ) -1 by low-dose aspirin leads to long-lasting suppression of thromboxane ( TX ) A2 production and TXA2-mediated platelet activation and aggregation . This effect is necessary and sufficient to explain aspirin 's unique ( among other P23219 inhibitors ) effectiveness in preventing atherothrombosis , as well as its shared ( with other antiplatelet agents ) bleeding liability . However , different mechanisms of action have been suggested to explain other beneficial effects of aspirin , such as prevention of venous thromboembolism , chemoprevention of colorectal ( and other ) cancers , and reduced risk of dementia . These mechanisms include acetylation of other proteins in blood coagulation , inhibition of P35354 activity , and other P36551 -independent mechanisms . The intent of this review is to develop the concept that the multifaceted therapeutic effects of low-dose aspirin may reflect pleiotropic consequences of platelet inhibition on pathophysiological tissue repair processes . Furthermore , the clinical implications of this concept will be discussed in terms of current clinical practice and future research . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Investigation of drug-drug interactions between clopidogrel and fluoxetine . Drug-drug interactions may contribute to the variability of the response of clopidogrel . Several hypotheses have been proposed concerning the potential modification of clopidogrel pharmacokinetics and pharmacodynamics by fluoxetine . This open-label crossover study assessed the effect of fluoxetine on the pharmacological activity of clopidogrel in healthy volunteers . Eight healthy male volunteers received a single 600-mg loading dose of clopidogrel followed by 20 mg of fluoxetine on 4 days and then 20 mg of fluoxetine plus 600 mg of clopidogrel on the fifth day . Eleven blood samples were withdrawn after clopidogrel administration to determine plasma concentrations of clopidogrel active metabolite ( P62158 ) and platelet function . Platelet aggregation was measured by light transmittance aggregometry ( P01374 ) and platelet reactivity index by flow cytometric vasodilator-stimulated phosphoprotein ( P50552 ) analysis . The areas under the curve and maximum plasma concentrations of P62158 were , respectively , 20.6 and 25.3 % lower after co-administration of fluoxetine compared with administration of clopidogrel alone . The percentage maximum platelet aggregation values in the presence of 5 μM and 10 μM adenosine diphosphate , measured by P01374 , were , respectively , 13.9 and 22.4 % lower after fluoxetine co-administration . The platelet reactivity index measured by the flow cytometric P50552 method was 36.8 % lower when clopidogrel was administered in conjunction with fluoxetine . Human platelets generate phospholipid-esterified prostaglandins via cyclooxygenase-1 that are inhibited by low dose aspirin supplementation . Oxidized phospholipids ( oxPLs ) generated nonenzymatically display pleiotropic biological actions in inflammation . Their generation by cellular cyclooxygenases ( COXs ) is currently unknown . To determine whether platelets generate prostaglandin ( PG ) -containing oxPLs , then characterize their structures and mechanisms of formation , we applied precursor scanning-tandem mass spectrometry to lipid extracts of agonist-activated human platelets . Thrombin , collagen , or ionophore activation stimulated generation of families of PGs comprising PGE₂ and D₂ attached to four phosphatidylethanolamine ( PE ) phospholipids ( 16:0p/ , 18:1p/ , 18:0p/ , and 18:0a/ ) . They formed within 2 to 5 min of activation in a calcium , phospholipase C , p38 Q96HU1 kinases , Q02750 , cPLA₂ , and src tyrosine kinase-dependent manner ( 28.1 ± 2.3 pg/2 × 10⁸ platelets ) . Unlike free PGs , they remained cell associated , suggesting an autocrine mode of action . Their generation was inhibited by in vivo aspirin supplementation ( 75 mg/day ) or in vitro P23219 blockade . Inhibitors of fatty acyl reesterification blocked generation significantly , while purified P23219 was unable to directly oxidize PE in vitro . This indicates that they form in platelets via rapid esterification of P23219 derived PGE₂/D₂ into PE . In summary , P23219 in human platelets acutely mediates membrane phospholipid oxidation via formation of PG-esterified PLs in response to pathophysiological agonists . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Genetic markers for differentiating aspirin-hypersensitivity . DB00945 -induced asthma ( AIA ) and aspirin-induced urticaria/ angioedema ( AIU ) are two major aspirin-related allergies . We summarize recent findings related to their molecular genetic mechanisms in order to identify genetic susceptibility markers for differentiating AIU and AIA . The overproduction of cysteinyl leukotriene has been suggested as a mechanism in both AIU and AIA . Increased expression of Q9Y271 with CYLSTR1 and Q9NS75 polymorphisms are new findings in AIA , while the P09917 promoter polymorphism has been noted in AIU . An HLA study suggested that DPB1*0301 is a strong genetic marker for AIA , and that HLA Q8IUH3 *1302 and DQB1*0609 are markers for AIU susceptibility . Several single nucleotide polymorphisms ( SNPs ) in the promoters of EP2 , Q9UL17 , P35354 , Fc epsilon RIbeta , and P21731 were associated with AIA , while an Fc epsilon RIalpha promoter polymorphism was associated with AIU . The functional studies of the key genes involved in AIA and AIU are summarized . The identification and functional study of genetic markers for AIA and AIU susceptibility would further elucidate the pathogenic mechanisms and facilitate the development of early diagnostic markers to establish therapeutic targets . Inhibition of P13671 rat glioma proliferation by [ Ru2Cl(Ibp)4 ] depends on changes in P38936 , p27 , Bax/Bcl2 ratio and mitochondrial membrane potential . The ruthenium compound [ Ru(2)Cl(Ibp)(4) ] ( or RuIbp ) has been reported to cause significantly greater inhibition of P13671 glioma cell proliferation than the parent HIbp . The present study determined the effects of 0-72h exposure to RuIbp upon P13671 cell cycle distribution , mitochondrial membrane potential , reactive species generation and mRNA and protein expression of Q01094 , cyclin D1 , c-myc , P06400 , P38936 , p27 , p53 , P12956 , P13010 , Bax , Bcl2 , cyclooxygenase 1 and 2 ( P23219 and P35354 ) . The most significant changes in mRNA and protein expression were seen for the cyclin-dependent kinase inhibitors P38936 and p27 which were both increased ( p < 0.05 ) . The marked decrease in mitochondrial membrane potential ( p < 0.01 ) and modest increase in apoptosis was accompanied by a decrease in anti-apoptotic Bcl2 expression and an increase in pro-apoptotic Bax expression ( p < 0.05 ) . Interestingly , P23219 expression was increased in response to a significant loss of prostaglandin E(2) production ( p < 0.001 ) , most likely due to the intracellular action of Ibp . Future studies will investigate the efficacy of this novel ruthenium-ibuprofen complex in human glioma cell lines in vitro and both rat and human glioma cells growing under orthotopic conditions in vivo . Epstein-Barr virus Zta-induced immunomodulators from nasopharyngeal carcinoma cells upregulate interleukin-10 production from monocytes . During lytic infection with Epstein-Barr virus ( EBV ) , several viral lytic proteins function to evade immune recognition or to actively suppress immune cells . An EBV lytic transactivator , Zta , induces an immunosuppressive cytokine interleukin 10 ( P22301 ) in B cells , but whether it regulates P22301 in the context of epithelial cells is unclear . In this study , we tested nasopharyngeal carcinoma ( NPC ) cell lines and found that Zta did not induce P22301 in these epithelial cells . Interestingly , the conditioned medium of Zta-expressing NPC cells enhanced P22301 production from monocytes . We further revealed that the P22301 -inducing effect involved at least two immunomodulators that were upregulated by Zta and secreted from NPC cells : granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) and prostaglandin E(2) ( PGE(2) ) . Zta was recruited to and activated the GM- P04141 promoter , thus upregulating GM- P04141 expression . Zta also activated the promoter of cyclooxygenase-2 ( P35354 ) , and Zta-induced P35354 increased downstream PGE(2) production . Cotreatment with GM- P04141 and PGE(2) synergistically induced P22301 production from monocytes . The P22301 -inducing effect of the Zta-conditioned medium was reduced when GM- P04141 or the P35354 /PGE(2) pathway was blocked . The conditioned medium of NPC cells with EBV lytic infection showed a similar increase of GM- P04141 and PGE(2) levels as well as the P22301 -inducing effect on monocytes , and knockdown of Zta abolished all the effects . Therefore , through Zta-induced immunomodulators , EBV lytic infection in NPC cells can direct bystander monocytes to produce P22301 , which may be a novel way of EBV to promote local immunosuppression . Effects of non-steroidal anti-inflammatory drugs on cyclo-oxygenase and lipoxygenase activity in whole blood from aspirin-sensitive asthmatics vs healthy donors . 1. Cyclo-oxygenase ( P36551 ) and lipoxygenase ( LO ) share a common substrate , arachidonic acid . DB00945 and related drugs inhibit P36551 activity . In a subset of patients with asthma aspirin induces clinical symptoms associated with increased levels of certain LO products , a phenomenon known as aspirin-sensitive asthma . The pharmacological pathways regulating such responses are not known . 2 . Here P23219 and LO activity were measured respectively by the formation of thromboxane B(2) ( TXB(2) ) or leukotrienes ( LT ) C(4) , D(4) and E(4) in whole blood stimulated with A23187 . P35354 activity was measured by the formation of prostaglandin E(2) ( PGE(2) ) in blood stimulated with lipopolysaccharide ( LPS ) for 18 h . 3 . No differences in the levels of P23219 , P35354 or LO products or the potency of drugs were found in blood from aspirin sensitive vs aspirin tolerant patients . DB00945 , indomethacin and nimesulide inhibited P23219 activity , without altering LO activity . Indomethacin , nimesulide and the P35354 selective inhibitor DB00677 [ 5,5-dimethyl-3-(2-isopropoxy)-4-(4-methanesulfonylphenyl)-2(5H)-furanone ] inhibited P35354 activity . NO-aspirin , like aspirin inhibited P23219 activity in blood from both groups . However , NO-aspirin also reduced LO activity in the blood from both patient groups . Sodium salicylate was an ineffective inhibitor of P23219 , P35354 or LO activity in blood from both aspirin-sensitive and tolerant patients . 4 . Thus , when P36551 activity in the blood of aspirin-sensitive asthmatics is blocked there is no associated increase in LO products . Moreover , NO-aspirin , unlike other NSAIDs tested , inhibited LO activity in the blood from both aspirin sensitive and aspirin tolerant individuals . This suggests that NO-aspirin may be better tolerated than aspirin by aspirin-sensitive asthmatics . Restriction of adenoviral replication to the transcriptional intersection of two different promoters for colorectal and pancreatic cancer treatment . In our current study , we developed oncolytic adenoviruses which preferentially lyse pancreatic and colon cancer cells by replacing viral E1 and/or E4 promoter with the tumor/tissue-specific promoters , cyclooxygenase-2 ( P35354 ) , midkine ( MK ) , or the cell cycle-dependent promoter , Q01094 . We generated three sets of recombinant adenoviral vectors . In the first set , only the native E1A promoter was replaced by the P35354 , MK , or Q01094 promoter , respectively . In the second set , the viral E4 promoter was substituted by these heterologous promoters and the viral E1A promoter was substituted by the ubiquitously active cytomegalovirus-IE promoter . In the third set , we substituted the viral E1A and E4 promoters with the P35354 , MK , or Q01094 promoter , respectively . In our system , transcriptional targeting of solitary viral E1A resulted in 50 % enhanced restricted vector replication when compared with an unrestricted replication-competent adenovirus . Furthermore , a targeted expression of the viral E1A gene products had a greater effect on restricted adenoviral replication than that of the E4 region . With our vectors , Ad. P36551 .MK and Ad.MK. P36551 , using two different heterologous promoters to control E1A and E4 expression , we showed enhanced viral replication specificity when compared with Ad. P36551 . P36551 or Ad.MK.MK , respectively . In a s.c. xenograft tumor model , there was no significant difference in the antineoplastic efficacy of the double heterologous promoter-controlled vectors when compared with our unrestricted replication-competent control adenovirus or vectors with only E1A transcriptionally driven by a heterologous promoter . DB08877 for the treatment of primary myelofibrosis . PURPOSE : The pharmacology , pharmacokinetics , pharmacogenomics , clinical efficacy , and safety profile of ruxolitinib for the treatment of primary myelofibrosis are reviewed . SUMMARY : DB08877 , an oral tyrosine kinase inhibitor that targets the Janus-associated kinases ( JAKs ) 1 and 2 , has been recently approved for the treatment of patients with intermediate- or high-risk myelofibrosis . Unlike previous treatment options for patients with myelofibrosis , ruxolitinib offers a targeted therapy option for these patients who often suffer with severe and debilitating symptoms associated with the disease process . After oral administration , ruxolitinib is rapidly absorbed and can be given without regard to meals . DB08877 is primarily metabolized by the cytochrome P-450 ( CYP ) 3A4 isoenzyme system ; therefore , if concomitant use with a strong P08684 inhibitor is unavoidable , an initial dosage reduction is warranted . Two Phase III randomized trials comparing ruxolitinib to either placebo or best available therapy found a rapid and sustained response in the reduction of spleen size and improvements in constitutional symptoms and quality of life , with one study demonstrating an improvement in overall survival . The most commonly reported serious adverse effects of ruxolitinib are anemia and thrombocytopenia . DB08877 is administered as an oral tablet given twice daily , with the initial starting dosage based on the baseline platelet count . Dosage reductions are based on the development of thrombocytopenia . CONCLUSION : By directly targeting both P23458 and O60674 through small-molecule inhibition , ruxolitinib elicits a reduction in splenomegaly and disease-related symptoms in patients with intermediate- or high-risk myelofibrosis while maintaining an acceptable toxicity profile and a low treatment-discontinuation rate . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity . [ DB00945 : recent advances in cardiovascular prevention ] . More than a century after being launched onto the market , aspirin still remains a fascinating drug , both for its demonstrated antalgic , antipyretic , antiinflammatory and antithrombotic properties and , also , for newer , yet conjectural , applications mentioned in recent publications . The role of aspirin , as an irreversible P23219 inhibitor and antiplatelet agent , is well elucidated and established . Our purpose is to review the value of aspirin for primary and secondary prevention of ischemic cardiovascular events . The clinician constantly has to manage a trade off between the protective effects of aspirin and its possible hemorrhagic , notably gastrointestinal , side-effects . The Task Force of the ESC recommends the use of doses no higher than 75-100 mg/d . New antiplatelet agents ( thienopyridin derivatives ) , which have a totally different mode of action , have been introduced and were compared with aspirin . Although clopidogrel may be slightly superior to the latter , according to the European experts : " the size of any additional benefit is statistically uncertain and the drug has not been granted a claim , of superiority " . Economical considerations reinforce this view . DB00758 is undoubtedly a good alternative when aspirin is contra-indicated , poorly tolerated , or not efficacious . Resistance to aspirin and resistance to clopidogrel have been described . In some high-risk patients , the combined use of aspirin and clopidogrel is deemed justified . DB00945 prevents apoptosis and NF-kappaB activation induced by H2O2 in hela cells . The classical pathway of nuclear factor-kappa B ( NF-kappaB ) activation by several inducers mainly involves the phosphorylation of P25963 by a signalsome complex composed of P25963 kinases ( IKKalpha and IKKbeta ) . However , in some cell types hydrogen peroxide ( H2O2 ) has been shown to activate an alternative pathway that does not involve the classical signalsome activation process . In this study , we demonstrate that H2O2 induced NF-kappaB activation in HeLa cells through phosphorylation and degradation of IkappaB proteins as shown by immunblot analysis . Our studies reveal that a commonly used non-steroid anti-inflammatory drug , acetylsalicylic acid ( aspirin ) prevents H2O2-induced NF-kappaB activation in a dose-dependent manner through inhibition of phosphorylation and degradation of P25963 and Q15653 . Differential staining and DNA fragmentation analysis also show that aspirin preloading of HeLa cells also prevents H2O2-induced apoptosis in a dose-dependent manner with maximum efficiency at 10 mM concentration . Additionally , aspirin effectively prevents caspase-3 and caspase-9 ( cysteinyl aspartate-specific proteases ) activation by H2O2 . These results suggest that NF-kappaB activation is involved in H2O2-induced apoptosis and aspirin may inhibit both processes simultaneously . De novo synthesis of cyclooxygenase-1 counteracts the suppression of platelet thromboxane biosynthesis by aspirin . DB00945 affords cardioprotection through the acetylation of serine529 in human cyclooxygenase-1 ( P23219 ) of anucleated platelets , inducing a permanent defect in thromboxane A2 ( TXA2 ) -dependent platelet function . However , heterogeneity of P23219 suppression by aspirin has been detected in cardiovascular disease and may contribute to failure to prevent clinical events . The recent recognized capacity of platelets to make proteins de novo paves the way to identify new mechanisms involved in the variable response to aspirin . We found that in washed human platelets , the complete suppression of TXA2 biosynthesis by aspirin , in vitro , recovered in response to thrombin and fibrinogen in a time-dependent fashion ( at 0.5 and 24 hours , TXB2 averaged 0.1+/-0.03 and 3+/-0.8 ng/mL ; in the presence of arachidonic acid [ 10 micromol/L ] , it was 2+/-0.7 and 25+/-7 ng/mL , respectively ) , and it was blocked by translational inhibitors , by rapamycin , and by inhibitors of phosphatidylinositol 3-kinase . The results that P23219 mRNA was readily detected in resting platelets and that [ 35S ] -methionine was incorporated into P23219 protein after stimulation strongly support the occurrence of de novo P23219 synthesis in platelets . This process may interfere with the complete and persistent suppression of TXA2 biosynthesis by aspirin necessary for cardioprotection . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . DB00945 inhibits P14780 mRNA expression and release via the PPARalpha/gamma and P35354 /mPGES-1-mediated pathways in macrophages derived from THP-1 cells . In present study , we investigated the effects of aspirin on matrix metalloproteinase ( MMP ) -9 mRNA expression and release and its possible mechanisms in macrophages derived from THP-1 cells . The macrophages were divided into different groups and treated with different drugs , the mRNA expression of P14780 , peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma , cyclooxygenase ( P36551 ) -2 , membranebound prostaglandin E synthase ( mPGES ) -1 in macrophages were examined with reverse-transcription polymerase chain reaction , and the protein expressions of Q07869 alpha and gamma , mPGES-1 were detected by Western-blot , the levels of P14780 and PGE(2) in cultured supernatants were determined with enzyme-linked immunosorbent assay . The results indicated that after the macrophages were incubated with aspirin for 24h , the P14780 mRNA expression and release were decreased , while the Q07869 alpha/gamma mRNA and protein expression was increased , respectively , and Q07869 alpha/gamma agonists could also decrease P14780 mRNA expression and release . Additionally , the P35354 mRNA expression , mPGES-1 mRNA and protein expression in macrophages were all decreased after incubation with aspirin for 24h and the PGE(2) release was also decreased . The macrophages stimulated with PGE(2) for 24h might increase the P14780 mRNA expression and release . When PGE(2) plus Q07869 alpha agonist or Q07869 gamma agonist were simultaneously used , the stimulation of P14780 mRNA expression and release by PGE(2) was significantly decreased . It might be concluded that aspirin could inhibit the P14780 gene expression and release through the PPARalpha/gamma and P35354 /mPGES-1-mediated pathways and the two pathways might be partly overlapped and even be interrelated . Effects of selective cyclooxygenase isoform inhibition on systemic prostacyclin synthesis and on platelet function at rest and after exercise in healthy volunteers . To test the hypothesis that selective inhibition of cyclooxygenase ( P36551 ) -2 would result in exercise-induced platelet activation by causing a shift in the endogenous thromboxane ( TX ) /prostacyclin balance , a double blind , randomized study comparing aspirin ( 300 mg/d ) with rofecoxib ( 25 mg/d ) ( cross-over design , 14 days washout between treatments ) in n = 10 trained healthy volunteers was carried out . Physical exercise resulted only in a minor platelet activation , as reflected by the expression of basal or ADP-stimulated platelet activation markers or basal plasma concentrations of TXB(2) . DB00945 significantly reduced TXB(2) in plasma while rofecoxib significantly increased TXB(2) in urine . Although no increase in systemic prostacyclin concentration was observed , there was a significant exercise-related increase in both platelet DB02527 and cGMP without any drug-related effects . It is concluded that , in trained healthy volunteers , selective inhibition of P23219 ( aspirin ) or P35354 ( rofecoxib ) does not affect systemic prostacyclin synthesis after physical exercise . However , our data do not exclude the possibility that in subjects at risk for atherothrombotic complications ( e.g. patients with advanced atherosclerotic disease ) P35354 inhibitors may result in platelet activation by inhibiting endothelial prostacyclin formation . Identification and absolute configuration of dihydroxy-arachidonic acids formed by oxygenation of 5S-HETE by native and aspirin-acetylated P35354 . Biosynthesis of the prostaglandin endoperoxide by the cyclooxygenase ( P36551 ) enzymes is accompanied by formation of a small amount of 11R-hydroxyeicosatetraenoic acid ( HETE ) , 15R-HETE , and 15S-HETE as by-products . Acetylation of P35354 by aspirin abrogates prostaglandin synthesis and triggers formation of 15R-HETE as the sole product of oxygenation of arachidonic acid . Here , we investigated the formation of by-products of the transformation of 5S-HETE by native P35354 and by aspirin-acetylated P35354 using HPLC-ultraviolet , GC-MS , and LC-MS analysis . 5S,15S- dihydroxy (di)HETE , 5S,15R-diHETE , and 5S,11R-diHETE were identified as by-products of native P35354 , in addition to the previously described di-endoperoxide ( 5S,15S-dihydroxy-9S,11R,8S,12S-diperoxy-6E,13E-eicosadienoic acid ) as the major oxygenation product . 5S,15R-diHETE was the only product formed by aspirin-acetylated P35354 . Both 5,15-diHETE and 5,11-diHETE were detected in Q86TM3 mouse colon carcinoma cells as well as in lipopolysaccharide-activated RAW264.7 cells incubated with 5S-HETE , and their formation was attenuated in the presence of the P35354 specific inhibitor , NS-398 . DB00945 -treated Q86TM3 cells gave 5,15-diHETE as the most prominent product formed from 5S-HETE . 5S,15S-diHETE has been described as a product of the cross-over of P09917 ( 5- P28300 ) and P16050 activities in elicited rat mononuclear cells and human leukocytes , and our studies implicate cross-over of the 5- P28300 and P35354 pathways as an additional biosynthetic route . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Safety of high-dose rofecoxib in patients with aspirin-exacerbated respiratory disease . BACKGROUND : DB00945 -exacerbated respiratory disease ( AERD ) is characterized by progressive sinusitis , nasal polyposis , and asthma that begins and continues in the absence of exposure to aspirin and nonsteroidal anti-inflammatory drugs ( NSAIDs ) . Cross-sensitivity to all NSAIDs that inhibit cyclooxygenase-1 ( P23219 ) occurs in these individuals . Reactions to aspirin and NSAIDs in patients with AERD are largely due to inhibition of P23219 . Despite accumulating data on the safety of P35354 selective inhibitors in AERD , concern still remains that high doses of a P35354 inhibitor may be sufficient to induce a cross-reaction . OBJECTIVE : To determine whether high-dose rofecoxib cross-reacts in patients with AERD and asthma . METHODS : Sixty asthmatic patients underwent blinded placebo-controlled oral challenges with 50 mg of rofecoxib . DB00945 sensitivity was subsequently confirmed in all patients with the use of single-blinded aspirin challenges . RESULTS : None of the 60 patients experienced any symptoms , changes in nasal examination results , or declines in lung function during rofecoxib challenge . All 60 patients experienced respiratory reactions to aspirin challenge , with a mean provoking dose of 57 mg . The exact 1-sided 95 % confidence interval for the underlying probability of 50 mg of rofecoxib inducing respiratory cross-reactions in patients with AERD is 0 to 0.05 , or 0 % to 5 % . CONCLUSIONS : These results confirm the lack of cross-reactivity of aspirin and the highly selective P35354 inhibitors in AERD . We suggest that it is time for the labeling of highly selective P35354 inhibitors to reflect these data and for the warning that patients with AERD in particular and asthmatic patients in general avoid selective P35354 inhibitors to be removed . P35354 -derived lipoxin A4 increases gastric resistance to aspirin-induced damage . BACKGROUND & AIMS : P35354 ( P35354 ) has been implicated as contributing to mucosal defense . Acetylation of P35354 by aspirin can result in production of an antiinflammatory substance , 15(R)-epi-LXA4 . We determined whether aspirin-triggered lipoxin ( LX ) production via P35354 diminishes aspirin-induced damage in the rat stomach . METHODS : Rats were treated with aspirin plus or minus celecoxib or rofecoxib . Gastric generation of LXA4 was measured . Effect of exogenous LXA4 or an P25090 antagonist on gastric resistance to aspirin-induced damage was examined . DB00945 -induced leukocyte adherence in mesenteric venules , and the effects of LXA4 , were examined by intravital microscopy . RESULTS : Celecoxib and rofecoxib significantly increased the severity of aspirin-induced gastric damage . DB00945 rapidly up-regulated P35354 expression in the stomach and caused a significant increase in gastric 15(R)-epi-LXA4 production , which was abolished by celecoxib . LXA4 dose dependently ( 0.25-2.5 microg/kg , intraperitoneally ) reduced the severity of aspirin-induced gastric damage and suppressed aspirin-induced leukocyte adherence , whereas an LXA4 antagonist had the opposite effects . CONCLUSIONS : DB00945 administration results in elevated production of 15(R)-epi-LXA4 via P35354 . LXA4 exerts very potent protective actions on the gastric mucosa . Co-administration of aspirin and a selective P35354 inhibitor results in substantially more severe gastric injury than is produced with either agent alone . [ Chemoprevention of colorectal cancer ] . Although colorectal cancer is one of the most preventable forms of cancer , it remains the second leading cause of cancer death worldwide . Primary prevention involves the identification and elimination of factors , which cause or promote colorectal cancer . The goal of screening is to prevent colorectal cancer mortality through the detection and treatment of premalignant adenomas and curable-stage cancer . Most colorectal cancers are believed to arise from adenomatous polyps . Early identification and removal of adenomas can prevent the development of colorectal cancer . Chemoprevention is the use of specific chemical compounds to prevent , inhibit , or reverse carcinogenesis . Several chemoprevention options have been investigated and confirmed as effective . DB00945 and other nonsteroidal anti-inflammatory drugs are the most widely studied agents , their use has been consistently associated with reduction in the risk of mortality and the incidence of colorectal adenomas and cancers . The selective cyclooxygenase-2 ( P35354 ) inhibitors ( coxibs ) have been demonstrated to decrease the number and the size of polyps in patients with familial adenomatous polyposis syndrome . Because the gastrointestinal toxicity of coxibs is lower , it might be safer than aspirin or other non-selective nonsteroidal anti-inflammatory drugs for long-term use . This review aims to summarize the recent theoretical and practical advances in the chemoprevention of colorectal cancer . Altered expression of beta-catenin , P12830 , cycloxygenase-2 , and p53 protein by ovine intestinal adenocarcinoma cells . Around 1.6 % of sheep in New Zealand develop small-intestinal adenocarcinomas . These neoplasms typically develop widespread metastases . The common development of these neoplasms and their subsequent behavior suggests that sheep could be a useful animal model of human colonic cancer . However , for an animal model of human disease to be relevant , similar genetic mutations should be present . Genetic mutations within human colonic cancers frequently result in expression of cycloxygenase-2 ( P35354 ) , loss of membranous expression of beta-catenin and P12830 , and accumulation of p53 protein within the neoplastic cells . Immunohistochemistry was used to investigate the presence of these 4 proteins within 26 ovine intestinal adenocarcinomas . Loss of membranous beta-catenin reactivity was observed in 14 of 26 ovine intestinal adenocarcinomas ( 54 % ) . The loss of membranous beta-catenin reactivity was accompanied by cytoplasmic and nuclear reactivity in 2 neoplasms . Loss of P12830 was observed within 8 of 26 neoplasms ( 31 % ) . Neoplastic cell expression of P35354 was observed in 12 of 26 neoplasms ( 46 % ) , whereas cells within 3 of 26 neoplasms ( 11 % ) contained visible p53 protein . In conclusion , all 4 proteins that commonly have altered expression in human colonic cancers were also altered in a proportion of the ovine intestinal adenocarcinomas . These results provide additional evidence that sheep could be useful for the study of human colonic cancer . [ Effects of danhong injection on experimental atherosclerosis rabbit model and its mechanism ] . OBJECTIVE : To observe the effects of Danhong injection on experimental atherosclerosis rabbits and explore its possible mechanism . METHODS : Forty New-Zealand male rabbits were randomly divided into four groups : normal control group , high-cholesterol group , danhong injection group and aspirin group , all rabbits were administered for fourteen weeks . The plaque area of aorta in each group was determined by image analysis . The blood lipid level was tested by ELISA method , malonaldehyde ( MDA ) in aorta wall was analyzed with thiobarbituric acid method , the P35228 and P35354 in aorta wall was tested with RT-PCR method . RESULT : There were significant difference between danhong injection group and other groups ( P < 0.5 ) , danhong injection and DB00945 decreased the TG , TC and LDL-C level in plasma of rabbit model and decreased the expression of inflammatory cytokines and the oxidative stress level in aorta wall . CONCLUSION : Danhong injection can inhibit the formation of experimental atherosclerosis in rabbits , which may be related to decreasing blood lipid , plaque inflammation and oxidative stress level . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Anti-inflammatory function of Withangulatin A by targeted inhibiting P35354 expression via MAPK and NF-kappaB pathways . Withangulatin A ( WA ) , an active component isolated from Physalis angulata L. , has been reported to possess anti-tumor and trypanocidal activities in model systems via multiple biochemical mechanisms . The aim of this study is to investigate its anti-inflammatory potential and the possible underlying mechanisms . In the current study , WA significantly suppressed mice T lymphocytes proliferation stimulated with LPS in a dose- and time-dependent manner and inhibited pro-inflammation cytokines ( P60568 , P01579 , and P05231 ) dramatically . Moreover , WA targeted inhibited P35354 expression mediated by MAPKs and NF-kappaB nuclear translocation pathways in mice T lymphocytes , and this result was further confirmed by the P23219 /2 luciferase reporter assay . Intriguingly , administration of WA inhibited the extent of mice ear swelling and decreased pro-inflammatory cytokines production in mice blood serum . Based on these evidences , WA influences the mice T lymphocytes function through targeted inhibiting P35354 expression via MAPKs and NF-kappaB nuclear translocation signaling pathways , and this would make WA a strong candidate for further study as an anti-inflammatory agent .

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interacts_with DB06603?

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Exploiting the TRIP-Br family of cell cycle regulatory proteins as chemotherapeutic drug targets in human cancer . Q9UHV2 and Q14140 are potent cell growth promoting factors that function as components of the Q01094 /DP1 transcription complex to integrate positive growth signals provided by P20941 zinc finger- and/or bromodomain-containing transcription factors . Q9UHV2 has been demonstrated to be an oncogene . We recently reported that antagonism of the TRIP-Br integrator function by synthetic decoy peptides that compete with TRIP-Br for binding to P20941 zinc finger- and/or bromodomain-containing proteins elicit an anti-proliferative effect and induces caspase-3-independent sub-diploidization in cancer cells in vitro . We now demonstrate the chemotherapeutic potential of TRIP-Br decoy peptides for the treatment of cutaneous and intracavitary lesions in vitro as well as in vivo in representative human nasopharyngeal cancer ( CNE2 ) , cervical cancer ( Ca Ski ) and melanoma ( MeWo ) cancer cell lines . In vitro , BrdU incorporation , colony formation assays and cell cycle analysis confirmed that TRIP-Br decoy peptides possess strong anti-proliferative effects and induce nuclear sub-diploidization in cancer cells . In vivo , CNE2 , Ca Ski and MeWo-derived chick embryo chorioallantoic membrane ( P62158 ) tumor xenografts were used to evaluate the effect of topically applied TRIP-Br peptides . Confocal microscopy and flow cytometric analysis demonstrated that cells comprising the tumor xenografts efficiently internalized topically applied FITC-labeled peptides . Fifty muM of Q9UHV2 decoy peptide significantly suppressed the growth of P61916 -derived human nasopharyngeal tumors , while 50 muM of Q14140 decoy peptide significantly inhibited tumor growth in all three P62158 tumor xenograft models . Two hundred muM of Q9UHV2 decoy peptide significantly inhibited MeWo-derived tumors . These results suggest that the TRIP-Br integrator function may represent a novel chemotherapeutic target for the treatment of human cutaneous and intracavitary proliferative lesions . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . Q8NFH3 / Q8TCB0 mitogen-activated protein kinases inhibit atrial natriuretic peptide mRNA transcription in P40189 -mediated hypertrophic ventricular myocytes . OBJECTIVE : To understand the role of P01160 mRNA transcription regulation in P40189 -mediated cardiomyocyte hypertrophy , and the involved mitogen-activated protein kinase kinase ( MEK ) -extracellular signal-regulated kinase ( P29323 , also called Q8NFH3 / Q8TCB0 MAPK ) signaling pathway . METHODS : Isolated neonatal ventricular myocytes were treated with different concentrations of Q16619 ( 10(-9) , 10(-8)and 10(-7)mol/L ) . MTT was used to analyze the viability and RT-PCR was used to detect P01160 mRNA levels in cardiomyocyte . To inhibit Q8NFH3 / Q8TCB0 MAPK activity in hypertrophic cardiomyocytes , the cells were pretreated with a specific Q02750 inhibitor . RESULTS : Q16619 significantly induced P01160 mRNA expression and the viability of cardiomyocytes in a dose- and time-dependent manner . Furthermore , blocking Q8NFH3 / Q8TCB0 MAPK activity by the special Q02750 inhibitor upregulated the P01160 mRNA . CONCLUSIONS : Q8NFH3 / Q8TCB0 MAPK have an important role in suppressing P01160 mRNA transcription and cell activity in P40189 -mediated hypertrophic ventricular myocytes . Comparison of the cytotoxicity of cladribine and clofarabine when combined with fludarabine and busulfan in AML cells : Enhancement of cytotoxicity with epigenetic modulators . Clofarabine ( Clo ) , fludarabine ( Flu ) , and busulfan ( Bu ) combinations are efficacious in hematopoietic stem cell transplantation for myeloid leukemia . We sought to determine whether the more affordable drug cladribine ( Clad ) can provide a viable alternative to Clo , with or without DB06603 ( Pano ) and DB01262 ( P22760 ) . Both Clad+Flu+Bu and Clo+Flu+Bu combinations showed synergistic cytotoxicity in KBM3/Bu250(6) , HL60 , and OCI- Q13950 cell lines . Cell exposure to these drug combinations resulted in 60 % -80 % inhibition of proliferation ; activation of the Q13315 pathway ; increase in histone modifications ; decrease in O15379 , P56524 , Q9UQL6 and SirT7 proteins ; decrease in mitochondrial membrane potential ; activation of apoptosis and stress signaling pathways ; and downregulation of the AKT pathway . These drug combinations activated DNA-damage response and apoptosis in primary cell samples from AML patients . At lower concentrations of Clad/Clo , Flu , and Bu , inclusion of Pano and P22760 enhanced cell killing , increased histone modifications and DNA demethylation , and increased the levels of P16/INK4a , P15/INK4b and P21/Waf1/Cip1 proteins . The observed DNA demethylating activity of Clad and Clo may complement P22760 activity ; increase demethylation of the gene promoters for Q8N474 , Q9UBP4 , and Q9Y5W5 ; and cause degradation of β-catenin in cells exposed to Clad/Clo+Flu+Bu+ P22760 +Pano . The overlapping activities of Clad/Clo+Flu+Bu , Pano , and P22760 in DNA-damage formation and repair , histone modifications , DNA demethylation , and apoptosis may underlie their synergism . Our results provide a basis for supplanting Clo with Clad and for including epigenetic modifiers in the pre-hematopoietic stem cell transplantation conditioning regimen for myeloid leukemia patients . Benzyl isothiocyanate ( BITC ) inhibits migration and invasion of human gastric cancer AGS cells via suppressing P29323 signal pathways . Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases . Benzyl isothiocyanate ( BITC ) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables . BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans . Here , the purpose of this study was to investigate the effects of BITC on cell proliferation , migration , invasion and mitogen-activated protein kinase ( MAPK ) pathways of AGS human gastric cancer cells . Wound healing and Boyden chamber ( migration and invasion ) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells . BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner . Results from Western blotting indicated that BITC exerted an inhibitory effect on the P27361 /2 , Ras , P62993 , Rho A , P35228 , P35354 for causing the inhibitions of P08253 , -7 and -9 then followed by the inhibitions of invasion and migration of AGS cells in vitro . BITC also promoted O14733 , Q99759 , c-jun , P45983 /2 , P15692 , Sos1 , phosphoinositide 3-kinase ( PI3K ) , PKC , nuclear factor-kappaB ( NF-κB ) p65 in AGS cells . Results from real-time polymerized chain reaction ( PCR ) showed that BITC inhibited the gene expressions of P08253 ,-7 -9 , Q05397 , Q13464 and RhoA after BITC treatment for 24 and 48 hours in AGS cells . Taken together , the finding may provide new mechanisms and functions of BITC , which inhibit migration and invasion of human gastric cancer AGS cells . Combined treatment with DB02546 , bortezomib , and clarithromycin for concomitant targeting of aggresome formation and intracellular proteolytic pathways enhances ER stress-mediated cell death in breast cancer cells . The ubiquitin-proteasome pathway and the autophagy-lysosome pathway are two major intracellular protein degradation systems . We previously reported that clarithromycin ( P62158 ) blocks autophagy flux , and that combined treatment with P62158 and proteasome inhibitor bortezomib ( BZ ) enhances ER-stress-mediated apoptosis in breast cancer cells , whereas treatment with P62158 alone results in almost no cytotoxicity . Since Q9UBN7 is involved in aggresome formation , which is recognized as a cytoprotective response serving to sequester misfolded proteins and facilitate their clearance by autophagy , we further investigated the combined effect of vorinostat ( suberoylanilide hydroxamic acid ( DB02546 ) ) , which has a potent inhibitory effect for Q9UBN7 , with P62158 and BZ in breast cancer cell lines . DB02546 exhibited some cytotoxicity along with an increased acetylation level of α-tubulin , a substrate of Q9UBN7 . Combined treatment of DB02546 , P62158 , and BZ potently enhanced the apoptosis-inducing effect compared with treatment using each reagent alone or a combination of two of the three . Expression levels of ER-stress-related genes , including the pro-apoptotic transcription factor P35638 ( P35638 ) , were maximally induced by the simultaneous combination of three reagents . Like breast cancer cell lines , a wild-type murine embryonic fibroblast ( MEF ) cell line exhibited enhanced cytotoxicity and maximally up-regulated Chop after combined treatment with DB02546 , P62158 , and BZ ; however , a Chop knockout MEF cell line almost completely canceled this enhanced effect . The specific Q9UBN7 inhibitor tubacin also exhibited a pronounced cytocidal effect with a combination of P62158 plus BZ . These data suggest that simultaneous targeting of intracellular proteolytic pathways and Q9UBN7 enhances ER-stress-mediated apoptosis in breast cancer cells . Histone deacetylase inhibitors as potential therapeutic approaches for chordoma : an immunohistochemical and functional analysis . Chordomas are rare malignancies of the axial skeleton . Therapy is mainly restricted to surgery . This study investigates histone deacetylase ( HDAC ) inhibitors as potential therapeutics for chordomas . Immunohistochemistry ( IHC ) was performed using the HDAC 1-6 antibodies on 50 chordoma samples ( 34 primary tumors , 16 recurrences ) from 44 patients ( 27 male , 17 female ) . Pan-HDAC-inhibitors DB02546 ( DB02546 ) , DB06603 ( LBH-589 ) , and Belinostat ( PXD101 ) were tested for their efficacy in the chordoma cell line MUG-Chor1 via Western blot , cell cycle analysis , caspase 3/7 activity ( MUG-Chor1 , UCh-1 ) , cleaved caspase-3 , and PARP cleavage . p-Values below 0.05 were considered significant . IHC was negative for Q13547 , positive for Q92769 in most ( n = 36 ; 72 % ) , and for HDACs 3-6 in all specimens available ( n = 43 ; 86 % ) . Q9UBN7 expression was strongest . DB02546 and LBH-589 , but not PXD101 caused a significant increase of G2/M phase cells and of cleaved caspase-3 ( p = 0.0003 , and p = 0.0014 after 72 h , respectively ) , and a peak of caspase 3/7 activity . PARP cleavage confirmed apoptosis . The presented chordoma series expressed HDACs 2-6 with strongest expression of Q9UBN7 . DB02546 and LBH-589 significantly increased apoptosis and changed cell cycle distribution in vitro . HDAC-inhibitors should be further evaluated as therapeutic options for chordoma . Role of acetylation and extracellular location of heat shock protein 90alpha in tumor cell invasion . Heat shock protein ( hsp ) 90 is an DB00171 -dependent molecular chaperone that maintains the active conformation of client oncoproteins in cancer cells . An isoform , hsp90alpha , promotes extracellular maturation of matrix metalloproteinase ( MMP ) -2 , involved in tumor invasion and metastasis . Knockdown of histone deacetylase ( HDAC ) 6 , which deacetylates lysine residues in hsp90 , induces reversible hyperacetylation and attenuates DB00171 binding and chaperone function of hsp90 . Here , using mass spectrometry , we identified seven lysine residues in hsp90alpha that are hyperacetylated after treatment of eukaryotic cells with a pan-HDAC inhibitor that also inhibits Q9UBN7 . Depending on the specific lysine residue in the middle domain involved , although acetylation affects DB00171 , cochaperone , and client protein binding to hsp90alpha , acetylation of all seven lysines increased the binding of hsp90alpha to 17-allyl-amino-demethoxy geldanamycin . Notably , after treatment with the pan-HDAC inhibitor DB06603 ( LBH589 ) , the extracellular hsp90alpha was hyperacetylated and it bound to P08253 , which was associated with increased in vitro tumor cell invasiveness . Treatment with antiacetylated hsp90alpha antibody inhibited in vitro invasion by tumor cells . Thus , reversible hyperacetylation modulates the intracellular and extracellular chaperone function of hsp90 , and targeting extracellular hyperacetylated hsp90alpha may undermine tumor invasion and metastasis . Pressure overload-induced cardiac hypertrophy response requires janus kinase 2-histone deacetylase 2 signaling . Pressure overload induces cardiac hypertrophy through activation of O60674 ( Jak2 ) , however , the underlying mechanisms remain largely unknown . In the current study , we tested whether histone deacetylase 2 ( Q92769 ) was involved in the process . We found that angiotensin II ( Ang-II ) -induced re-expression of fetal genes ( Atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) ) in cultured cardiomyocytes was prevented by the Jak2 inhibitor AG-490 and Q92769 inhibitor Trichostatin-A ( P32119 ) , or by Jak2/ Q92769 siRNA knockdown . On the other hand , myocardial cells with Jak2 or Q92769 over-expression were hyper-sensitive to Ang-II . In vivo , pressure overload by transverse aorta binding ( AB ) induced a significant cardiac hypertrophic response as well as re-expression of P01160 and DB04899 in mice heart , which were markedly reduced by AG-490 and P32119 . Significantly , AG-490 , the Jak2 inhibitor , largely suppressed pressure overload-/Ang-II-induced Q92769 nuclear exportation in vivo and in vitro . Meanwhile , P32119 or Q92769 siRNA knockdown reduced Ang-II-induced P01160 / DB04899 expression in Jak2 over-expressed H9c2 cardiomyocytes . Together , these results suggest that Q92769 might be a downstream effector of Jak2 to mediate cardiac hypertrophic response by pressure overload or Ang-II . Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors . The human HDAC ( histone deacetylase ) family , a well-validated anticancer target , plays a key role in the control of gene expression through regulation of transcription . While HDACs can be subdivided into three main classes , the class I , class II and class III HDACs ( sirtuins ) , it is presently unclear whether inhibiting multiple HDACs using pan-HDAC inhibitors , or targeting specific isoforms that show aberrant levels in tumours , will prove more effective as an anticancer strategy in the clinic . To address the above issues , we have tested a number of clinically relevant HDACis ( HDAC inhibitors ) against a panel of rhHDAC ( recombinant human HDAC ) isoforms . Eight rhHDACs were expressed using a baculoviral system , and a Fluor de Lystrade mark ( Biomol International ) HDAC assay was optimized for each purified isoform . The potency and selectivity of ten HDACs on class I isoforms ( rhHDAC1 , rhHDAC2 , rhHDAC3 and rhHDAC8 ) and class II HDAC isoforms ( rhHDAC4 , rhHDAC6 , rhHDAC7 and rhHDAC9 ) was determined . MS-275 was Q13547 -selective , DB05651 was Q13547 - and Q92769 -selective , apicidin was Q92769 - and O15379 -selective and valproic acid was a specific inhibitor of class I HDACs . The hydroxamic acid-derived compounds ( trichostatin A , DB00238 -LAQ824 , DB06603 , ITF2357 , vorinostat and belinostat ) were potent pan-HDAC inhibitors . The growth-inhibitory effect of the HDACis on HeLa cells showed that both pan-HDAC and class-I-specific inhibitors inhibited cell growth . The results also showed that both pan-HDAC and class-I-specific inhibitor treatment resulted in increased acetylation of histones , but only pan-HDAC inhibitor treatment resulted in increased tubulin acetylation , which is in agreement with their activity towards the Q9UBN7 isoform . Q9UBN7 gates the synaptic action of acute stress in prefrontal cortex . The prefrontal cortex ( P27918 ) , a region responsible for high-order cognitive functions , such as decision-making , attention and working memory , is highly influenced by stress and corticosteroid stress hormones . Recently it has been shown that acute stress affects P27918 functions by potentiating glutamatergic transmission via a mechanism dependent on glucocorticoid receptor ( GR ) and its downstream target , serum and glucocorticoid-inducible kinase ( O00141 ) . To identify the key regulators of stress responses , we examined the role of histone deacetylase 6 ( Q9UBN7 ) , a unique member of the HDAC family that could regulate the GR chaperone protein heat shock protein 90 ( HSP90 ) , in the synaptic action of acute stress in P27918 . We found that Q9UBN7 inhibition or knockdown blocked the enhancement of glutamatergic transmission and glutamate receptor trafficking by acute stress in vivo or corticosterone treatment in vitro . In addition , Q9UBN7 inhibition blocked the up-regulation of O00141 in animals exposed to acute stress . HSP90 inhibition or knockdown produced a similar blockade of the acute stress-induced enhancement of glutamatergic signalling . These findings have identified Q9UBN7 as a key molecule gating the effects of acute stress on synaptic functions in the P27918 . Identification of an amino acid residue important for binding of methiothepin and sumatriptan to the human 5-HT(1B) receptor . Site-directed mutagenesis of the human P28222 receptor was performed to investigate the role of the amino acid residues cysteine 326 and tryptophan 327 in transmembrane region VI and aspartic acid 352 in transmembrane region VII in ligand binding . Binding studies were performed with the antagonist radioligand [3H]GR125743 on mutant and wild-type receptors stably expressed in Chinese hamster ovary cells ( CHO ) - P04264 cells . Substitution of tryptophan 327 by alanine resulted in decreased affinities of all ligands tested . The most prominent changes in affinity were observed for the antagonist methiothepin and the antimigraine drug sumatriptan , which were reduced approximately 300- and 60-fold , respectively . Nevertheless , the affinity of 5-HT remained the same . Replacement of the aspartic acid 352 by alanine reduced high-affinity binding of 5-HT . Substitution of cysteine 326 by alanine had minor effects on ligand binding . Some of these results agree with the results from mutagenesis studies of the corresponding amino acids in other receptors . However , some notable differences also emerge showing that functional roles of individual amino acid residues must be tested experimentally in each receptor subtype . Effects of histone deacetylase inhibitors on amygdaloid histone acetylation and neuropeptide Y expression : a role in anxiety-like and alcohol-drinking behaviours . Recent studies have demonstrated the involvement of epigenetic mechanisms in psychiatric disorders , including alcoholism . Here , we investigated the effects of histone deacetylase ( HDAC ) inhibitor , trichostatin A ( P32119 ) on amygdaloid HDAC-induced histone deacetylation and neuropeptide Y ( P01303 ) expression and on anxiety-like and alcohol-drinking behaviours in alcohol-preferring ( P ) and -non-preferring ( NP ) rats . It was found that P rats displayed higher anxiety-like and alcohol-drinking behaviours , higher amygdaloid nuclear , but not cytosolic , HDAC activity , which was associated with increased Q92769 protein levels and deficits in histone acetylation and P01303 expression in the central ( CeA ) and medial nucleus of amygdala ( MeA ) , as compared to NP rats . P32119 treatment attenuated the anxiety-like and alcohol-drinking behaviours , with concomitant reductions in amygdaloid nuclear , but not cytosolic HDAC activity , and Q92769 , but not P56524 , protein levels in the CeA and MeA of P rats , without effect in NP rats . P32119 treatment also increased global histone acetylation ( H3- P35527 and H4-K8 ) and P01303 expression in the CeA and MeA of P , but not in NP rats . Histone H3 acetylation within the P01303 promoter was also innately lower in the amygdala of P rats compared with NP rats ; which was normalized by P32119 treatment . Voluntary ethanol intake in P , but not NP rats , produced anxiolytic effects and decreased the Q92769 levels and increased histone acetylation in the CeA and MeA . These results suggest that higher Q92769 expression-related deficits in histone acetylation may be involved in lower P01303 expression in the amygdala of P rats , and operative in controlling anxiety-like and alcohol-drinking behaviours . Interactions of alpha- , beta- , and theta-defensins with influenza A virus and surfactant protein D . We have reported that the alpha-defensins human neutrophil peptides ( HNP ) -1 and HNP-2 neutralize and aggregate influenza A virus ( IAV ) and promote uptake of IAV by neutrophils . These alpha-defensins were also shown to bind to surfactant protein ( SP ) -D and reduce its antiviral activity . In this study , we examined retrocyclin (RC)1 and RC2 , humanized versions of the antiviral theta-defensins found in the leukocytes of certain nonhuman primates . Q9HC52 was just as effective as P59665 -3 in neutralizing IAV , and RC2 and RC101 ( an analog of Q9HC52 ) were more effective . In contrast , human beta-defensins ( HBDs ) showed less neutralizing activity . Human defensins 5 and 6 ( mainly produced by intestinal Paneth cells ) had viral neutralizing activity similar to P59665 -3 . Like P59665 -3 , RCs induced viral aggregation and promoted the uptake of IAV by neutrophils . We used surface plasmon resonance to evaluate binding of defensins to P35247 . HBDs , Q9UBN7 , and P12838 bound minimally to P35247 . P59665 -3 and RCs bound P35247 with high affinity ; however , unlike P59665 and HNP-2 , RCs did not inhibit P35247 antiviral activity . HBDs also did not inhibit antiviral activity of P35247 . Given their strong neutralizing activity and compatibility with P35247 , RCs may provide attractive prototypes for designing therapeutics that can prevent or treat respiratory infections caused by IAV . P10912 targeting to lipid rafts requires extracellular subdomain 2 . P10912 ( P10912 ) is a single membrane-spanning glycoprotein dimer that binds GH in its extracellular domain ( O95905 ) . GH activates the P10912 intracellular domain ( ICD ) -associated tyrosine kinase , O60674 , which causes intracellular signaling . We previously found that plasma membrane ( PM ) -associated P10912 was dramatically enriched in the lipid raft ( LR ) component of the membrane and that localization of P10912 within PM regions may regulate GH signaling by influencing the profile of pathway activation . In this study , we examined determinants of LR localization of the P10912 using a reconstitution system which lacks endogenous O60674 and P10912 . By non-detergent extraction and multistep fractionation , we found that P10912 was highly enriched in the LR fraction independent of O60674 expression . Various P10912 mutants were examined in transfectants harboring O60674 . LR concentration was observed for a P10912 in which the native transmembrane domain ( TMD ) is replaced by that of the unrelated P01130 and for a P10912 that lacks its ICD . Thus , LR association requires neither the TMD nor the ICD . Similarly , a P10912 that lacks the O95905 , except for the membrane-proximal O95905 stem region , was only minimally LR-concentrated . Mutants with internal stem deletions in the context of the full-length receptor were LR-concentrated similar to the wild-type . A P10912 lacking O95905 subdomain 1 reached the PM and was LR-concentrated , while one lacking O95905 subdomain 2 , also reached the PM , but was not LR-concentrated . These data suggest LR targeting resides in O95905 subdomain 2 , a region relatively uninvolved in GH binding . c- P14921 facilitates P55008 /S-phase transition by up-regulating cyclin E and P24941 genes and cooperates with hepatitis B virus X protein for their deregulation . Recent studies on the molecular mechanisms responsible for cell cycle deregulation in cancer have puzzled out the role of oncogenes in mediating unscheduled cellular proliferation . This is reminiscence of their activity as proto-oncogenes that drives scheduled cell cycle progression under physiological conditions . Working on the cell cycle regulatory activity of proto-oncogene , we observed that c- P14921 transcriptionally up-regulated both cyclin E and P24941 genes , the master regulators of G(1)/S-phase transition . The process was mediated by kinetic coherence of c- P14921 expression and its recruitment to both promoters during G(1)/S-phase transition . Furthermore , enforced expression of c- P14921 helped G(0)-arrested cells to progress into G(1)/S-phases apparently due to the activation of cyclin E/ P24941 genes . Physiological induction of c- P14921 by P01133 showed the remodeling of mononucleosomes bound to the c- P14921 binding site on both promoters during their activation . The exchange of Q13547 with histone acetyltransferase-p300 was contemporaneous to the chromatin remodeling with consequent increase in histone H3K9 acetylation . Furthermore , the DB00171 -dependent chromatin remodeler hBRM1 recruitment was also associated with nucleosome remodeling and promoter occupancy of phospho-Ser5 RNA polymerase II . Intriguingly , the activity of the HBx viral oncoprotein was dependent on c- P14921 in a hepatotropic manner , which led to the activation of cyclin E/ P24941 genes . Thus , cyclin E and P24941 genes are key physiological effectors of the c- P14921 proto-oncogene . Furthermore , c- P14921 is indispensable for the hepatotropic action of HBx in cell cycle deregulation . Activation of the JAK/ P35610 pathway in vascular smooth muscle by serotonin . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a vasoconstrictor and mitogen whose levels are elevated in diabetes . Previous studies have shown the presence of 5- Q13049 , P41595 , and P28222 receptors in vascular smooth muscle cells ( VSMCs ) . There are currently no data regarding P41595 and P28222 receptor activation of the JAK/ P35610 pathway in VSMCs and resultant potential alterations in 5-HT signaling in diabetes . Therefore , we tested the hypothesis that 5-HT differentially activates the JAK/ P35610 pathway in VSMCs under conditions of normal ( 5 mM ) and high ( 25 mM ) glucose . Treatment of rat VSMCs with 5-HT ( 10(-6) M ) resulted in time-dependent activation ( approximately 2-fold ) of O60674 , P23458 , and P42224 , but not P40763 ( maximal at 5 min , returned to baseline by 30 min ) . The P41595 receptor agonist BW723C86 and the P28222 receptor agonist CGS12066A ( 10(-9)-10(-5) M , 5-min stimulation ) did not activate the JAK/ P35610 pathway . Treatment with the 5- Q13049 receptor antagonist ketanserin ( 10 nM ) inhibited O60674 activation by 5-HT . Treatment of streptozotocin-induced diabetic rats with ketanserin ( 5 mg.kg-1.day-1 ) reduced activation of O60674 and P42224 but not P40763 in endothelium-denuded thoracic aorta in vivo . 5-HT ( 10(-6) M ) treatment resulted in increased cell proliferation and increased DNA synthesis , which were inhibited by the O60674 inhibitor AG490 . Further studies with apocynin , diphenyleneiodonium chloride , catalase , and virally transfected superoxide dismutase had no effect at either glucose concentration on activation of the JAK/ P35610 pathway by 5-HT . Therefore , we conclude that 5-HT activates O60674 , P23458 , and P42224 via the 5- Q13049 receptors in a reactive oxygen species-independent manner under both normal and high glucose conditions . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Histone deacetylase inhibitor treatment dramatically reduces cholesterol accumulation in Niemann-Pick type C1 mutant human fibroblasts . Niemann-Pick type C ( NPC ) disease is predominantly caused by mutations in the O15118 protein that affect intracellular cholesterol trafficking and cause accumulation of unesterified cholesterol and other lipids in lysosomal storage organelles . We report the use of a series of small molecule histone deacetylase ( HDAC ) inhibitors in tissue culture models of NPC human fibroblasts . Some HDAC inhibitors lead to a dramatic correction in the NPC phenotype in cells with either one or two copies of the O15118 (I1061T) mutation , and for several of the inhibitors , correction is associated with increased expression of O15118 protein . Increased O15118 (I1061T) protein levels may partially account for the correction of the phenotype , because this mutant can promote cholesterol efflux if it is delivered to late endosomes and lysosomes . The HDAC inhibitor treatment is ineffective in an P61916 mutant human fibroblast line . Analysis of the isoform selectivity of the compounds used implicates Q13547 and/or Q92769 as likely targets for the observed correction , although other HDACs may also play a role . LBH589 ( DB06603 ) is an orally available HDAC inhibitor that crosses the blood-brain barrier and is currently in phase III clinical trials for several types of cancer . It restores cholesterol homeostasis in cultured O15118 mutant fibroblasts to almost normal levels within 72 h when used at 40 nM . The findings that HDAC inhibitors can correct cholesterol storage defects in human O15118 mutant cells provide the potential basis for treatment options for NPC disease . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Both the ADP receptors P47900 and Q9H244 , play a role in controlling shape change in human platelets . Two types of ADP receptors , P2Y(1) and P2Y(12) are involved in platelet aggregation . The P2X(1) receptor is also present but its role , in terms of platelet function , is not yet defined . The aim of this study was to establish if the ADP receptors , P2Y ( 1 , ) P2Y(12) and P2X(1) play a role in controlling platelet shape change ( PSC ) in human platelets . PSC is an early phase of platelet activation that precedes aggregation . Using a high-resolution channelyzer , PSC was assessed by measuring the median platelet volume ( MPV ) . The P2Y(1) receptor antagonist P59665 2179 ( 1.06 - 10.25 micro mol/l ) blocked ADP-induced PSC ( by 100 % ) . The median IC(50) was 3.16 micro mol/l . P59665 2179 also significantly ( P = 0.01 ) inhibited PSC induced by the combination of ADP + serotonin ( 5HT ) . The P2Y(12) receptor antagonist AR-C69931MX significantly inhibited ( at 10s , P = 0.009 ; 15 s , P = 0.001 and 30 s , P = 0.015 ) ADP-induced PSC . The P2X(1) receptor antagonist TNP- DB00171 had no significant effect on ADP- or ADP + 5HT-induced PSC . We conclude that the IC(50) of a P2Y(1)-blocker can be derived because of the high-resolution and reproducibility of the channelyzer technique . In addition to the P2Y(1) purinoceptor , the P2Y(12)receptor appears to be involved in ADP-induced PSC since this process was significantly inhibited by AR-C69931MX . The channelyzer technique may be more reliable than optical aggregometry to assess PSC . Combination therapy for hepatocellular carcinoma : additive preclinical efficacy of the HDAC inhibitor DB06603 with sorafenib . BACKGROUND & AIMS : Hepatocellular carcinoma ( HCC ) is a heterogeneous cancer in which sorafenib is the only approved systemic therapy . Histone deacetylases ( HDAC ) are commonly dysregulated in cancer and therefore represent promising targets for therapies , however their role in HCC pathogenesis is still unknown . We analyzed the expression of 11 HDACs in human HCCs and assessed the efficacy of the pan-HDAC inhibitor DB06603 alone and in combination with sorafenib in preclinical models of liver cancer . METHODS : Gene expression and copy number changes were analyzed in a cohort of 334 human HCCs , while the effects of DB06603 and sorafenib were evaluated in three liver cancer cell lines and a murine xenograft model . RESULTS : Aberrant HDAC expression was identified and validated in 91 and 243 HCCs , respectively . Upregulation of O15379 and Q9UQL6 mRNAs was significantly correlated with DNA copy number gains . Inhibiting HDACs with DB06603 led to strong anti-tumoral effects in vitro and vivo , enhanced by the addition of sorafenib . Cell viability and proliferation declined , while apoptosis and autophagy increased . DB06603 increased histone H3 and HSP90 acetylation , downregulated O15392 ( survivin ) and upregulated CDH1 . Combination therapy with DB06603 and sorafenib significantly decreased vessel density , and most significantly decreased tumor volume and increased survival in HCC xenografts . CONCLUSIONS : Aberrant expression of several HDACs and copy number gains of O15379 and Q9UQL6 occur in HCC . Treatment with DB06603 combined with sorafenib demonstrated the highest preclinical efficacy in HCC models , providing the rationale for clinical studies with this novel combination . Comparative analyses of gene copy number and mRNA expression in glioblastoma multiforme tumors and xenografts . Development of model systems that recapitulate the molecular heterogeneity observed among glioblastoma multiforme ( GBM ) tumors will expedite the testing of targeted molecular therapeutic strategies for GBM treatment . In this study , we profiled DNA copy number and mRNA expression in 21 independent GBM tumor lines maintained as subcutaneous xenografts ( GBMX ) , and compared GBMX molecular signatures to those observed in GBM clinical specimens derived from the Cancer Genome Atlas ( TCGA ) . The predominant copy number signature in both tumor groups was defined by chromosome-7 gain/chromosome-10 loss , a poor-prognosis genetic signature . We also observed , at frequencies similar to that detected in TCGA GBM tumors , genomic amplification and overexpression of known GBM oncogenes , such as P00533 , Q00987 , Q00534 , and P04198 , and novel genes , including P57740 , Q7Z769 , P03956 , P45452 , and Q92499 . The transcriptional signature of GBMX tumors , which was stable over multiple subcutaneous passages , was defined by overexpression of genes involved in M phase , DNA replication , and chromosome organization ( MRC ) and was highly similar to the poor-prognosis mitosis and cell-cycle module ( P22033 ) in GBM . Assessment of gene expression in TCGA-derived GBMs revealed overexpression of MRC cancer genes Q96GD4 , O15392 , P14635 , O95067 , P06493 , P24941 , and Q08050 , which form a transcriptional network important for G2/M progression and/or checkpoint activation . Our study supports propagation of GBM tumors as subcutaneous xenografts as a useful approach for sustaining key molecular characteristics of patient tumors , and highlights therapeutic opportunities conferred by this GBMX tumor panel for testing targeted therapeutic strategies for GBM treatment . [ Effect of down-regulation of histone deacetylase 2 protein expression on cell proliferation and cell cycle in cervical carcinoma ] . OBJECTIVE : To study the effect of down-regulation of histone deacetylase 2 ( Q92769 ) expression on cell proliferation and cell cycle in cervical carcinoma cell lines HeLa . METHODS : Q92769 siRNA and control siRNA were transfected to HeLa cells . CCK-8 and flow cytometry were used to analyze the changes of cell proliferation and cell cycle , respectively . Western blot was employed to detect the changes of cell proliferation and cell cycle-related proteins . RESULTS : Q92769 siRNA significantly down-regulated the expression of Q92769 protein in HeLa cells , resulting in marked inhibition of cell proliferation . In addition , the percentage of cells in G(0)/G(1) phase in Q92769 siRNA group ( 63.3 % ± 2.0 % ) was significantly higher than that in untreated group ( 29.3 % ± 1.7 % ) or control siRNA group ( 29.4 % ± 1.7 % ) , F = 354.181 , P = 0.000 . Furthermore , Western blot demonstrated that down-regulation of Q92769 expression decreased the expression of cyclin D1 , cyclin E and P24941 proteins but increased the expression of P38936 protein . CONCLUSIONS : Down-regulation of Q92769 expression mediates proliferation inhibition and cell cycle arrest . It is associated with decrease in cyclin D1 , cyclin E and P24941 protein expression and increase in P38936 protein expression . Q9UBN7 sustains growth stimulation by prolonging the activation of P01133 receptor through the inhibition of rabaptin-5-mediated early endosome fusion in gastric cancer . The aberrant regulation of histone deacetylase 6 ( Q9UBN7 ) contributes to malignant progression in various types of cancer , but the mechanism underlying gastric carcinogenesis remains unknown . Aberrant Q9UBN7 overexpression was observed in a subset of human gastric cancer cells . Q9UBN7 knockdown caused the significant inhibition of gastric cancer cell growth without affecting the transition of cell cycles or the processing of cell death . We demonstrate that an increase in epidermal growth factor receptor ( P00533 ) signaling through decreased P00533 degradation was mediated by Q9UBN7 in gastric carcinogenesis . These results establish a molecular mechanism responsible for oncogenic Q9UBN7 , explaining how P00533 signaling induced by the growth factor is sustained during the malignant progression of gastric cancer . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Regulation of P40763 by histone deacetylase-3 in diffuse large B-cell lymphoma : implications for therapy . Diffuse large B-cell lymphoma ( DLBCL ) with an activated B-cell ( DB01048 ) gene-expression profile has been shown to have a poorer prognosis compared with tumors with a germinal center B-cell type . ABC cell lines have constitutive activation of P40763 ; however , the mechanisms regulating P40763 signaling in lymphoma are unknown . In studies of class-I histone deacetylase ( HDAC ) expression , we found overexpression of O15379 in phospho P40763 -positive DLBCL and the O15379 was found to be complexed with P40763 . Inhibition of HDAC activity by DB06603 ( LBH589 ) increased p300-mediated P40763 (Lys685) acetylation with increased nuclear export of P40763 to the cytoplasm . HDAC inhibition abolished P40763 (Tyr705) phosphorylation with minimal effect on P40763 (Ser727) and O60674 tyrosine activity . pSTAT3(Tyr705)-positive DLBCLs were more sensitive to HDAC inhibition with LBH589 compared with pSTAT3(Tyr705)-negative DLBCLs . This cytotoxicity was associated with downregulation of the direct P40763 target Mcl-1 . O15379 knockdown upregulated P40763 (Lys685) acetylation but prevented P40763 (Tyr705) phosphorylation and inhibited survival of pSTAT3-positive DLBCL cells . These studies provide the rationale for targeting P40763 -positive DLBCL tumors with HDAC inhibitors . The HDAC inhibitor LBH589 induces P29323 -dependent prometaphase arrest in prostate cancer via Q9UBN7 inactivation and down-regulation . Histone deacetylase inhibitors ( HDACIs ) have potent anti-cancer activity in a variety of cancer models . Understanding the molecular mechanisms involved in the therapeutic responsiveness of HDACI is needed before its clinical application . This study aimed to determine if a potent HDACI , LBH589 ( DB06603 ) , had differential therapeutic responsiveness towards LNCaP and PC-3 prostate cancer ( PCa ) cells . The former showed prometaphase arrest with subsequent apoptosis upon LBH589 treatment , while the latter was less sensitive and had late G2 arrest . The LBH589 treatment down-regulated Q9UBN7 and sustained P29323 activation , and contributed to prometaphase arrest . Mechanistically , LBH589 inhibited Q9UBN7 activity , caused its dissociation from protein phosphatase PP1α , and increased 14-3-3ζ acetylation . Acetylated 14-3-3ζ released its mask effect on serine 259 of c-Raf and serine 216 of Cdc25C subsequent to de-phosphorylation by PP1α , which contributed to P29323 activation . Enhanced P29323 activity by LBH589 further down-regulated Q9UBN7 protein levels and sustained P29323 activation by free-forward regulation . The sustained Cdc25C and P29323 activation resulted in early M-phase ( prometaphase ) arrest and subsequent apoptosis in the most sensitive LNCaP cells but not in PC-3 cells . This study provides pre-clinical evidence that Q9UBN7 may serve as a sensitive therapeutic target in the treatment of prostate cancer with HDACI LBH589 for clinical translation . This study also posits a novel mechanism of Q9UBN7 participation in regulating the c-Raf- P50391 - P29323 signaling pathway and contributing to M phase cell-cycle transition . Deciphering the molecular and biologic processes that mediate histone deacetylase inhibitor-induced thrombocytopenia . Histone deacetylase inhibitor ( HDACI ) -induced thrombocytopenia ( DB01520 ) is a major dose-limiting toxicity of this new class of drugs . Using preclinical models to study the molecular and biologic events that underpin this effect of HDACI , we found that C57BL/6 mice treated with both the Q13547 /2-selective HDACI romidepsin and the pan-HDACI DB06603 developed significant DB01520 . HDACI-induced DB01520 was not due to myelosuppression or reduced platelet lifespan , but to decreased platelet release from megakaryocytes . Cultured primary murine megakaryocytes showed reductions in proplatelet extensions after HDACI exposure and a dose-dependent increase in the phosphorylation of myosin light chain 2 ( MLC2 ) . Phosphorylation of MLC to phospho-MLC ( pMLC ) and subsequent proplatelet formation in megakaryocytes is regulated by the Rho-GTPase proteins Rac1 , P60953 , and RhoA . Primary mouse megakaryocytes and the human megakaryoblastic cell line Meg-01 showed reductions in Rac1 , P60953 , and RhoA protein levels after treatment with HDACIs . We were able to overcome HDACI-induced DB01520 by administering the mouse-specific thrombopoietin ( P07202 ) mimetic AMP-4 , which improved platelet numbers to levels similar to untreated controls . Our report provides the first detailed account of the molecular and biologic processes involved in HDACI-mediated DB01520 . Moreover , our preclinical studies provide evidence that dose-limiting DB01520 induced by HDACIs may be circumvented using a P07202 mimetic . Glioma cell activation by Alzheimer 's peptide Abeta1-42 , alpha1-antichymotrypsin , and their mixture . We compared the effects ofAlzheimer 's peptide ( Abeta1-42 ) , a,-antichymotrypsin ( ACT ) and an ACT/Abeta1-42 mixture on human glioma DK-MG cells . The solution of Abeta ( 5 microM ) formed by 2-h incubation at room temperature induced tumour necrosis factor-alpha ( P01375 ) and interleukin ( IL ) -6 levels by 55 and 45 % , respectively , and increased gelatinase B activity by 67 % , while exposure of cells to the ACT/Abeta1-42 mixture ( 1:10 molar ratio ACT : Abeta1-42 ) under the same experimental conditions showed no effect on P05231 levels or gelatinase B activity , but strongly induced P01375 ( by 190 % ) , compared to the controls . Stimulation of the cells with Abeta1-42 alone , but not with ACT , increased by about 20 % low-density lipoprotein ( LDL ) uptake and mRNA levels for P01130 and P04035 , while the ACT/Abeta1-42 mixture significantly increased LDL uptake ( by 50 % ) , up-regulated mRNA levels for P01130 and P04035 by 48 and 63 % , respectively , and increased lipid accumulation by about 20-fold . These data suggest a possible new role for Abeta in Alzheimer 's disease through its interaction with the inflammatory reactant , ACT . P05231 ( P05231 ) and/or soluble P05231 receptor down-regulation of human type II collagen gene expression in articular chondrocytes requires a decrease of Sp1.Sp3 ratio and of the binding activity of both factors to the P02458 promoter . Type II collagen is composed of alpha1(II) chains encoded by the P02458 gene . Alteration of this cartilage marker is a common feature of osteoarthritis . P05231 ( P05231 ) is a pro-inflammatory cytokine that needs a soluble form of receptor called sIL-6R to exert its effects in some cellular models . In that case , sIL-6R exerts agonistic action . This mechanism can make up for the partial or total absence of membrane-anchored P05231 receptors in some cell types , such as chondrocytes . Our study shows that P05231 , sIL-6R , or both inhibit type II collagen production by rabbit articular chondrocytes through a transcriptional control . The cytokine and/or sIL-6R repress P02458 transcription by a -63/-35 sequence that binds Sp1.Sp3 . Indeed , P05231 and/or sIL-6R inhibit Sp1 and Sp3 expression and their binding activity to the 63-bp promoter . In chromatin immunoprecipitation experiments , P05231 .sIL-6R induced an increase in Sp3 recruitment to the detriment of Sp1 . Knockdown of Sp1.Sp3 by small interference RNA and decoy strategies were found to prevent the P05231 - and/or sIL-6R-induced inhibition of P02458 transcription , indicating that each of these Sp proteins is required for down-regulation of the target gene and that a heterotypic Sp1.Sp3 complex is involved . Additionally , Sp1 was shown to interact with Sp3 and Q13547 . Indeed , overexpression of a full-length Sp3 cDNA blocked the Sp1 up-regulation of the 63-bp P02458 promoter activity , and by itself , inhibits P02458 transcription . We can conclude that P05231 , sIL-6R , or both in combination decrease both the Sp1.Sp3 ratio and DNA-binding activities , thus inhibiting P02458 transcription . Statins control oxidized LDL-mediated histone modifications and gene expression in cultured human endothelial cells . OBJECTIVE : Activation of the endothelium by oxidized low-density lipoprotein ( oxLDL ) has been implicated in the development of atherosclerosis . Histone modifications impact on the transcriptional activity state of genes . We tested the hypothesis that oxLDL-induced inflammatory gene expression is regulated by histone modifications and experienced the effect of statins on these alterations . METHODS AND RESULTS : OxLDL-related interleukin-8 ( P10145 ) and monocyte-chemoattractant protein-1 ( P13500 ) secretion in endothelial cells was reduced by statins but enhanced by histone deacetylase inhibitors . OxLDL induced lectin-like oxidized P01130 -1 ( P78380 ) and extracellular regulated kinases ( P27361 /2 ) -dependent acetylation of histone H3 and H4 as well as phosphorylation of histone H3 , both globally and on the promoters of il8 and mcp1 . Pretreatment of oxLDL-exposed cells with statins reduced the above mentioned histone modification , as well as recruitment of CREB binding protein ( CBP ) 300 , NF-kappaB , and of RNA polymerase II but prevented loss of binding of histone deacetylase ( HDAC ) -1 and -2 at the il8 and mcp1 gene promoters . OxLDL reduced Q13547 and 2 expression , and statins partly restored global HDAC-activity . Statin-related effects were reverted with mevalonate . In situ experiments indicated decreased expression of Q92769 in endothelial cells in atherosclerotic plaques of human coronary arteries . CONCLUSIONS : Histone modifications seem to play an important role in atherosclerosis . Methylphenidate and atomoxetine enhance prefrontal function through α2-adrenergic and dopamine D1 receptors . OBJECTIVE : This study examined the effects of the attention-deficit/hyperactivity disorder treatments , methylphenidate ( DB00422 ) and atomoxetine ( Q13315 ) , on prefrontal cortex ( P27918 ) function in monkeys and explored the receptor mechanisms underlying enhancement of P27918 function at the behavioral and cellular levels . METHOD : Monkeys performed a working memory task after administration of a wide range of DB00422 or Q13315 doses . The optimal doses were challenged with the α(2)-adrenoceptor antagonist , idazoxan , or the D(1) dopamine receptor antagonist , SCH23390 ( P35240 ) . In a parallel physiology study , neurons were recorded from the dorsolateral P27918 of a monkey performing a working memory task . Q13315 , P35240 , or the α(2) antagonist , yohimbine , were applied to the neurons by iontophoresis . RESULTS : DB00422 and Q13315 generally produced inverted-U dose-response curves , with improvement occurring at moderate doses , but not at higher doses . The beneficial effects of these drugs were blocked by idazoxan or P35240 . At the cellular level , Q13315 produced an inverted-U dose-response effect on memory-related firing , enhancing firing for preferred directions ( increasing " signals " ) and decreasing firing for the nonpreferred directions ( decreasing " noise " ) . The increase in persistent firing for the preferred direction was blocked by yohimbine , whereas the suppression of firing for the nonpreferred directions was blocked by P35240 . CONCLUSIONS : Optimal doses of DB00422 or Q13315 improved P27918 cognitive function in monkeys . These enhancing effects appeared to involve indirect stimulation of α(2) adrenoceptors and D(1) dopamine receptors in the P27918 . These receptor actions likely contribute to their therapeutic effects in the treatment of attention-deficit/hyperactivity disorder . Ontogenesis and regulation of cholesterol metabolism in the central nervous system of the mouse . These studies characterized the ontogenesis and regulation of cholesterol turnover in the central nervous system ( CNS ) of mice . During the first 3 weeks after birth , the CNS grew rapidly and equaled 5 % of body weight . The cholesterol pool in this tissue expanded at a rate of 0.26 mg/day and the CNS synthesized sterol at a rate of 0.28 mg/day . In mature mice between 13 and 26 weeks of age , there was a marked decrease in these parameters including a reduction in the relative size of the CNS to 1.7 % of body weight , a decrease in the rate of sterol accretion to 0.012 mg/day , and a reduction in the rate of cholesterol synthesis to 0.035 mg/day . Deletion of the O15118 and Q9Y6A2 proteins markedly altered cholesterol metabolism in the CNS . However , changes in the plasma cholesterol concentration or loss of function of DB00171 -binding cassette AI transporter ( O95477 ) , scavenger receptor class B , type I ( Q8WTV0 ) , low-density lipoprotein receptor ( P01130 ) , P02649 or APOAI had no effect on sterol turnover in the brain . Thus , during early development , cholesterol comes entirely from local synthesis . In the adult , however , synthesis exceeds the need for structural cholesterol so that there is a constant excretion of sterol from the CNS into the plasma at a rate of about 0.023 mg/day . P35354 regulates the proliferation of glioma stem like cells . Cancer stem-like cells ( CSCs ) possessing features of neural precursor cells ( NPC ) influence initiation , recurrence and chemoresistance of glioblastoma multiforme ( GBM ) . As inflammation is crucial for glioblastoma progression we investigated the effect of chronic IL-1β treatment on CSCs derived from glioblastoma cell line U87MG . Exposure to IL-1β for 10 days increased ( i ) accumulation of 8-OHdG - a key biomarker of oxidative DNA damage ; ( ii ) DNA damage response ( DDR ) indicators γ P16104 , Q13315 and DNA-PK ; ( iii ) nuclear and cytoplasmic p53 and P35354 levels and ( iv ) interaction between P35354 and p53 . Despite upregulating p53 expression IL-1β had no effect on cell cycle progression , apoptosis or self renewal capacity of CSCs . P35354 inhibitor Celecoxib reduced self renewal capacity and increased apoptosis of both control and IL-1β treated CSCs . Therefore the ability of P35354 to regulate proliferation of CSCs irrespective of exposure to IL-1β , warrants further investigation of P35354 as a potential anti-glioma target . The microtubule-associated histone deacetylase 6 ( Q9UBN7 ) regulates epidermal growth factor receptor ( P00533 ) endocytic trafficking and degradation . Q9UBN7 ( Q9UBN7 ) is a microtubule-associated deacetylase with tubulin deacetylase activity , and it binds dynein motors . Recent studies revealed that microtubule acetylation affects the affinity and processivity of microtubule motors . These unique properties implicate a role for Q9UBN7 in intracellular organelle transport . Here , we show that Q9UBN7 associates with the endosomal compartments and controls epidermal growth factor receptor ( P00533 ) trafficking and degradation . We found that loss of Q9UBN7 promoted P00533 degradation . Mechanistically , Q9UBN7 deficiency did not cause aberrant P00533 internalization and recycling . Rather , it resulted in accelerated segregation of P00533 from early endosomes and premature delivery of P00533 to the late endosomal and lysosomal compartments . The deregulated P00533 endocytic trafficking was accompanied by an increase in microtubule-dependent movement of P00533 -bearing vesicles , revealing a novel regulation of P00533 vesicular trafficking and degradation by the microtubule deacetylase Q9UBN7 . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . YAP modifies cancer cell sensitivity to P00533 and survivin inhibitors and is negatively regulated by the non-receptor type protein tyrosine phosphatase 14 . The Yes-associated protein ( YAP ) is a transcriptional factor involved in tissue development and tumorigenesis . Although YAP has been recognized as a key element of the Hippo signaling pathway , the mechanisms that regulate YAP activities remain to be fully characterized . In this study , we demonstrate that the non-receptor type protein tyrosine phosphatase 14 ( Q15678 ) functions as a negative regulator of YAP . We show that YAP forms a protein complex with Q15678 through the WW domains of YAP and the PPXY motifs of Q15678 . In addition , Q15678 inhibits YAP-mediated transcriptional activities . Knockdown of YAP sensitizes cancer cells to various anti-cancer agents , such as cisplatin , the P00533 tyrosine kinase inhibitor erlotinib and the small-molecule antagonist of survivin , P28222 . YAP-targeted modalities may be used in combination with other cancer drugs to achieve maximal therapeutic effects . Mechanistic insights into the events that lead to synergistic induction of interleukin 6 transcription upon activation of the aryl hydrocarbon receptor and inflammatory signaling . The aryl hydrocarbon receptor ( P35869 ) is the ligand-activated transcription factor responsible for mediating the toxicological effects of dioxin and xenobiotic metabolism . However , recent evidence has implicated the P35869 in additional , nonmetabolic physiological processes , including immune regulation . Certain tumor cells are largely nonresponsive to cytokine-mediated induction of the pro-survival cytokine interleukin ( IL ) 6 . We have demonstrated that multiple nonresponsive tumor lines are able to undergo synergistic induction of P05231 following combinatorial treatment with IL1beta and the P35869 agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin . Such data implicate the P35869 in tumor expansion , although the mechanistic basis for the P35869 -dependent synergistic induction of P05231 has not been determined . Here , we demonstrate that ligand-activated P35869 is involved in priming the P05231 promoter through binding to nonconsensus dioxin response elements located upstream of the P05231 start site . Such binding appears to render the promoter more permissive to IL1beta-induced binding of NF-kappaB components . The nature of the P35869 -dependent increases in P05231 promoter transcriptional potential has been shown to involve a reorganization of repressive complexes as exemplified by the presence of Q13547 and O15379 . Dismissal of these HDACs correlates with post-translational modifications of promoter-bound NF-kappaB components in a time-dependent manner . Thus the P35869 plays a role in derepressing the P05231 promoter , leading to synergistic P05231 expression in the presence of inflammatory signals . These observations may explain the association between enhanced expression of P35869 and tumor aggressiveness . It is likely that P35869 -mediated priming is not restricted to the P05231 promoter and may contribute to the expression of a variety of genes , which do not have consensus dioxin response elements . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Treatment with DB06603 induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells . Increased levels of misfolded polypeptides in the endoplasmic reticulum ( ER ) triggers the dissociation of glucose-regulated protein 78 ( P11021 ) from the three transmembrane ER-stress mediators , i.e. , protein kinase RNA-like ER kinase ( Q9NZJ5 ) , activating transcription factor-6 ( P18850 ) , and inositol-requiring enzyme 1alpha , which results in the adaptive unfolded protein response ( UPR ) . In the present studies , we determined that histone deacetylase-6 ( Q9UBN7 ) binds and deacetylates P11021 . Following treatment with the pan-histone deacetylase inhibitor DB06603 ( Novartis Pharmaceuticals ) , or knockdown of Q9UBN7 by short hairpin RNA , P11021 is acetylated in 11 lysine residues , which dissociates P11021 from Q9NZJ5 . This is associated with the activation of a lethal UPR in human breast cancer cells . Coimmunoprecipitation studies showed that binding of Q9UBN7 to P11021 requires the second catalytic and COOH-terminal BUZ domains of Q9UBN7 . Treatment with DB06603 increased the levels of phosphorylated-eukaryotic translation initiation factor ( p-eIF2alpha ) , P18848 , and CAAT/enhancer binding protein homologous protein ( P35638 ) . DB06603 treatment also increased the proapoptotic Q13323 , O43521 , Q07812 , and Q16611 levels , as well as increased the activity of caspase-7 . Knockdown of P11021 sensitized MCF-7 cells to bortezomib and DB06603 -induced UPR and cell death . These findings indicate that enforced acetylation and decreased binding of P11021 to Q9NZJ5 is mechanistically linked to DB06603 -induced UPR and cell death of breast cancer cells . Mol Cancer Ther ; 9(4) ; 942-52 . (c)2010 AACR . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Prefrontal cortical dysfunction after overexpression of histone deacetylase 1 . BACKGROUND : Postmortem brain studies have shown that Q13547 -a lysine deacetylase with broad activity against histones and nonhistone proteins-is frequently expressed at increased levels in prefrontal cortex ( P27918 ) of subjects diagnosed with schizophrenia and related disease . However , it remains unclear whether upregulated expression of Hdac1 in the P27918 could affect cognition and behavior . METHODS : Using adeno-associated virus , an Hdac1 transgene was expressed in young adult mouse P27918 , followed by behavioral assays for working and long-term memory , repetitive activity , and response to novelty . Prefrontal cortex transcriptomes were profiled by microarray . Antipsychotic drug effects were explored in mice treated for 21 days with haloperidol or clozapine . RESULTS : Hdac1 overexpression in P27918 neurons and astrocytes resulted in robust impairments in working memory , increased repetitive behaviors , and abnormal locomotor response profiles in novel environments . Long-term memory remained intact . Over 300 transcripts showed subtle but significant changes in Hdac1-overexpressing P27918 . Major histocompatibility complex class II ( MHC II ) -related transcripts , including HLA-DQA1/H2-Aa , P01920 /H2-Ab1 , and HLA- Q8IUH3 /H2-Eb1 , located in the chromosome 6p21.3-22.1 schizophrenia and bipolar disorder risk locus , were among the subset of genes with a more robust ( > 1.5-fold ) downregulation in expression . Hdac1 levels declined during the course of normal P27918 development . Antipsychotic drug treatment , including the atypical clozapine , did not affect Hdac1 levels in P27918 but induced expression of multiple MHC II transcripts . CONCLUSIONS : Excessive Q13547 activity , due to developmental defects or other factors , is associated with behavioral alterations and dysregulated expression of MHC II and other gene transcripts in the P27918 . DB06603 synergizes with bortezomib to induce endoplasmic reticulum stress and ubiquitinated protein accumulation in renal cancer cells . BACKGROUND : Inducing endoplasmic reticulum ( ER ) stress is a novel strategy used to treat malignancies . Inhibition of histone deacetylase ( HDAC ) 6 by the HDAC inhibitor DB06603 hinders the refolding of unfolded proteins by increasing the acetylation of heat shock protein 90 . We investigated whether combining DB06603 with the proteasome inhibitor bortezomib would kill cancer cells effectively by inhibiting the degradation of these unfolded proteins , thereby causing ubiquitinated proteins to accumulate and induce ER stress . METHODS : Caki-1 , ACHN , and 769-P cells were treated with DB06603 and/or bortezomib . Cell viability , clonogenicity , and induction of apoptosis were evaluated . The in vivo efficacy of the combination was evaluated using a murine subcutaneous xenograft model . The combination-induced ER stress and ubiquitinated protein accumulation were assessed . RESULTS : The combination of DB06603 and bortezomib induced apoptosis and inhibited renal cancer growth synergistically ( combination indexes < 1 ) . It also suppressed colony formation significantly ( p < 0.05 ) . In a murine subcutaneous tumor model , a 10-day treatment was well tolerated and inhibited tumor growth significantly ( p < 0.05 ) . Enhanced acetylation of the Q9UBN7 substrate alpha-tubulin was consistent with the suppression of Q9UBN7 activity by DB06603 , and the combination was shown to induce ER stress and ubiquitinated protein accumulation synergistically . CONCLUSIONS : DB06603 inhibits renal cancer growth by synergizing with bortezomib to induce ER stress and ubiquitinated protein accumulation . The current study provides a basis for testing the combination in patients with advanced renal cancer . Intraplatelet signaling mechanisms of the priming effect of matrix metalloproteinase-2 on platelet aggregation . OBJECTIVE : Platelets contain and release some matrix metalloproteinases ( MMPs ) , enzymes involved in the degradation of extracellular matrix , and one of these ( P08253 ) exerts a proaggregatory effect . We explored the signal transduction mechanisms activated by P08253 in human blood platelets . METHODS AND RESULTS : Recombinant , human P08253 , added before stimulation with subthreshold doses of different agonists , potentiated platelet activation , calcium influx , IP3 formation , and pleckstrin phosphorylation . Wortmannin and LY29400 , two P19957 -K inhibitors , suppressed the potentiating effects of P08253 and preincubation with P08253 enhanced the thrombin-induced association of the p85alpha P19957 -K subunit with the cytoskeleton and increased the phosphorylation of P31749 . Protein tyrosine kinase inhibitors , Q96HU1 kinase inhibitors , P04054 inhibitors , cyclooxygenase inhibitors and antagonists of the P47900 and Q9H244 receptors did not affect the potentiating activity of P08253 on platelets . CONCLUSION : Our data show that P08253 , at a concentration released by activated platelets , facilitates platelet activation acting at the level of a second messenger system common to different agonists and related to the activation of P19957 -K . Platelet-released P08253 may contribute to platelet activation in vivo . Discovery and validation of methylation markers for endometrial cancer . The prognosis of endometrial cancer is strongly associated with stage at diagnosis , suggesting that early detection may reduce mortality . Women who are diagnosed with endometrial carcinoma often have a lengthy history of vaginal bleeding , which offers an opportunity for early diagnosis and curative treatment . We performed DNA methylation profiling on population-based endometrial cancers to identify early detection biomarkers and replicated top candidates in two independent studies . We compared DNA methylation values of 1,500 probes representing 807 genes in 148 population-based endometrial carcinoma samples and 23 benign endometrial tissues . Markers were replicated in another set of 69 carcinomas and 40 benign tissues profiled on the same platform . Further replication was conducted in The Cancer Genome Atlas and in prospectively collected endometrial brushings from women with and without endometrial carcinomas . We identified 114 CpG sites showing methylation differences with p values of ≤ 10(-7) between endometrial carcinoma and normal endometrium . Eight genes ( P18509 , Q99929 , Q9Y278 , P28222 , MME , P01303 and O00570 ) were selected for further replication . Age-adjusted odds ratios for endometrial cancer ranged from 3.44 ( 95 % -CI : 1.33-8.91 ) for Q99929 to 18.61 ( 95 % -CI : 5.50-62.97 ) for P28222 . An area under the curve ( AUC ) of 0.93 was achieved for discriminating carcinoma from benign endometrium . Replication in The Cancer Genome Atlas and in endometrial brushings from an independent study confirmed the candidate markers . This study demonstrates that methylation markers may be used to evaluate women with abnormal vaginal bleeding to distinguish women with endometrial carcinoma from the majority of women without malignancy . O15379 acts as a negative regulator of angiogenesis . Histone deacetylase-3 ( O15379 ) is involved in cellular proliferation , apoptosis and transcriptional repression . However , the role of O15379 in angiogenesis remains unknown . O15379 negatively regulated the expression of angiogenic factors , such as P15692 and plasminogen activator inhibitor-1 ( P05121 ) . O15379 showed binding to promoter sequences of P05121 . O15379 activity was necessary for the expression regulation of P05121 by O15379 . P15692 decreased the expression of O15379 , and the down-regulation of O15379 enhanced endothelial cell tube formation . O15379 negatively regulated tumor-induced angiogenic potential . We show the novel role of O15379 as a negative regulator of angiogenesis . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene : interactive roles of modified histones , histone acetyltransferase , p300 , AND Sp1 . Atrial natriuretic peptide ( P01160 ) binds guanylyl cyclase-A/natriuretic peptide receptor-A ( P16066 /NPRA ) and produces the intracellular second messenger , cGMP , which regulates cardiovascular homeostasis . We sought to determine the function of histone deacetylases ( HDACs ) in regulating Npr1 ( coding for P16066 /NPRA ) gene transcription , using primary mouse mesangial cells treated with class-specific HDAC inhibitors ( HDACi ) . Trichostatin A , a pan inhibitor , and mocetinostat ( DB05651 ) , a class I HDAC inhibitor , significantly enhanced Npr1 promoter activity ( by 8- and 10-fold , respectively ) , mRNA levels ( 4- and 5.3-fold , respectively ) , and NPRA protein ( 2.7- and 3.5-fold , respectively ) . However , MC1568 ( class II HDAC inhibitor ) had no discernible effect . Overexpression of Q13547 and Q92769 significantly attenuated Npr1 promoter activity , whereas O15379 and Q9BY41 had no effect . HDACi-treated cultured cells in vitro and intact animals in vivo showed significantly reduced binding of Q13547 and -2 and increased accumulation of acetylated H3- P35527 /14 and H4-K12 at the Npr1 promoter . Deletional analyses of the Npr1 promoter along with ectopic overexpression and inhibition of Sp1 confirmed that HDACi-induced Npr1 gene transcription is accomplished by Sp1 activation . Furthermore , HDACi attenuated the interaction of Sp1 with Q13547 /2 and promoted Sp1 association with p300 and p300/ DB02527 -binding protein-associated factor ; it also promoted the recruitment of p300 and p300/ DB02527 -binding protein-associated factor to the Npr1 promoter . Our results demonstrate that trichostatin A and DB05651 enhanced Npr1 gene expression through inhibition of Q13547 /2 and increased both acetylation of histones ( H3- P35527 /14 , H4-K12 ) and Sp1 by p300 , and their recruitment to Npr1 promoter . Our findings define a novel epigenetic regulatory mechanism that governs Npr1 gene transcription . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Investigation of the hub genes and related mechanism in ovarian cancer via bioinformatics analysis . BACKGROUND : Ovarian cancer is a cancerous growth arising from the ovary . OBJECTIVE : This study was aimed to explore the molecular mechanism of the development and progression of the ovarian cancer . METHODS : We first identified the differentially expressed genes ( DEGs ) between the ovarian cancer samples and the healthy controls by analyzing the GSE14407 affymetrix microarray data , and then the functional enrichments of the DEGs were investigated . Furthermore , we constructed the protein-protein interaction network of the DEGs using the STRING online tools to find the genes which might play important roles in the progression of ovarian cancer . In addition , we performed the enrichment analysis to the PPI network . RESULTS : Our study screened 659 DEGs , including 77 up- and 582 down-regulated genes . These DEGs were enriched in pathways such as Cell cycle , p53 signaling pathway , Pathways in cancer and Drug metabolism . P24864 , O95067 and P20815 were the significant genes identified from these pathways . Protein-protein interaction ( PPI ) network was constructed and network Module A was found closely associated with ovarian cancer . Hub nodes such as P15692 , P62158 , O15392 and P28340 were found in the PPI network . Module A was related to biological processes such as mitotic cell cycle , cell cycle , nuclear division , and pathways namely Cell cycle , Oocyte meiosis and p53 signaling pathway . CONCLUSIONS : It indicated that ovarian cancer was closely associated to the dysregulation of p53 signaling pathway , drug metabolism , tyrosine metabolism and cell cycle . Besides , we also predicted genes such as P24864 , O95067 , P20815 and P15692 might be target genes for diagnosing the ovarian cancer . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . R306465 is a novel potent inhibitor of class I histone deacetylases with broad-spectrum antitumoral activity against solid and haematological malignancies . R306465 is a novel hydroxamate-based histone deacetylase ( HDAC ) inhibitor with broad-spectrum antitumour activity against solid and haematological malignancies in preclinical models . R306465 was found to be a potent inhibitor of Q13547 and -8 ( class I ) in vitro . It rapidly induced histone 3 ( H3 ) acetylation and strongly upregulated expression of p21waf1,cip1 , a downstream component of Q13547 signalling , in A2780 ovarian carcinoma cells . R306465 showed class I HDAC isotype selectivity as evidenced by poor inhibition of Q9UBN7 ( class IIb ) confirmed by the absence of downregulation of Hsp90 chaperone c-raf protein expression and tubulin acetylation . This distinguished it from other HDAC inhibitors currently in clinical development that were either more potent towards Q9UBN7 ( e.g. vorinostat ) or had a broader HDAC inhibition spectrum ( e.g. DB06603 ) . R306465 potently inhibited cell proliferation of all main solid tumour indications , including ovarian , lung , colon , breast and prostate cancer cell lines , with IC50 values ranging from 30 to 300 nM . Haematological cell lines , including acute lymphoblastic leukaemia , acute myeloid leukaemia , chronic lymphoblastic leukaemia , chronic myeloid leukaemia , lymphoma and myeloma , were potently inhibited at a similar concentration range . R306465 induced apoptosis and inhibited angiogenesis in cell-based assays and had potent oral in vivo antitumoral activity in xenograft models . Once-daily oral administration of R306465 at well-tolerated doses inhibited the growth of A2780 ovarian , H460 lung and HCT116 colon carcinomas in immunodeficient mice . The high activity of R306465 in cell-based assays and in vivo after oral administration makes R306465 a promising novel antitumoral agent with potential applicability in a broad spectrum of human malignancies . Q8IXB1 is the ER reductase that catalyzes the removal of non-native disulfides and correct folding of the P01130 . Q8IXB1 is a member of the protein disulfide isomerase family of proteins localized to the endoplasmic reticulum ( ER ) of mammalian cells . To date , only a limited number of substrates for Q8IXB1 are known . Here we identify a number of endogenous substrates that form mixed disulfides with Q8IXB1 , greatly expanding its client repertoire . Q8IXB1 previously had been thought to exclusively reduce disulfides in proteins destined for dislocation to the cytosol for degradation . However , we demonstrate here that for one of the identified substrates , the low-density lipoprotein receptor ( P01130 ) , Q8IXB1 is required not for degradation , but rather for efficient folding . Our results demonstrate that the crucial role of Q8IXB1 is to reduce non-native disulfides formed during productive folding and that this requirement is dependent on its interaction with P11021 . Hence , Q8IXB1 acts as the ER reductase , both preparing misfolded proteins for degradation and catalyzing the folding of proteins that form obligatory non-native disulfides . Epigenetics , an early event in the modulation of gene expression by inositol hexaphosphate in ethylnitrosourea exposed mouse lungs . Mechanisms of anticancer effects of inositol hexaphosphate are not fully understood . Epigenetic changes are the early changes in tumorigenesis . DNA methyl transferases , methyl CpG binding proteins , methyl CpG DNA binding domain protein , and histone deacetylases are the major molecules involved in epigenetics . We have shown the effects of IP6 at the molecular level in mouse lungs before the tumor is developed . After 3 mo of ENU exposure , there was no tumor formation , but there was hyperplasia and lymphocytic infiltration in the lungs . Inflammation and DNA damage repair enzymes P35354 and P40692 appear to be upregulated , whereas tumor suppressor gene p16 was downregulated by ENU . On the other hand , ENU exposure more or less upregulated the epigenetic events such as the expressions of P26358 , MeCP2 , Q9UIS9 , and Q13547 . This alteration was reduced by IP6 administration . Results were supported by modulation of global DNA methylation and the modulation of promoter CpG methylation of p16 , P40692 , and P35354 genes . Hence , this study indicates the possible role of epigenetics at the early stage of tumor development and in the regulation of gene expression by IP6 before the onset of ENU-induced lung tumors . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Molecular analysis of mutated thyroid peroxidase detected in patients with total iodide organification defects . Wild-type and mutant thyroid peroxidase ( P07202 ) was expressed in a Semliki Forest Virus ( SFV ) -based transient expression system in Chinese hamster ovary- P04264 cells . Twenty four hours after transfection proteins immunoreactive with P07202 antibodies could be detected on a Western blot . Peroxidase activity was assayed using both the guaiacol and the I3- assay . Addition of hematin was necessary to obtain enzymatic active P07202 . P07202 complementary DNA constructs containing mutations originally found in patients with hereditary congenital hypothyroidism caused by total iodide organification defects were analyzed using these techniques . In all cases P07202 was expressed as shown by Western blotting and immunostaining . Enzymatic activity ( measured by guaiacol and iodide oxidation assay ) was below the detection level in four out of five mutants . The only mutant yielding P07202 with enzymatic activity was G 1858 A ( DB00145 590 DB00133 ) . However , the mutation could affect splicing of P07202 messenger RNA , leading to inactive P07202 , because it is located at the exon 10/intron 10 border .

MH_train_1091

MH_train_1091

MH_train_1091

interacts_with DB00013?

multiple_choice

Korean Red DB01404 Suppresses Metastasis of Human Hepatoma SK-Hep1 Cells by Inhibiting Matrix Metalloproteinase-2/-9 and DB00013 P00747 Activator . Korean red ginseng and ginsenosides have been claimed to possess wide spectrum of medicinal effects , of which anticancer effect is one . The present study was undertaken to investigate the antimetastatic effect of Korean red ginseng on human hepatoma as well as possible mechanisms . The inhibitory effect of the water extract of Korean red ginseng ( WKRG ) on the invasion and motility of SK-Hep1 cells was evaluated by the Boyden chamber assay in vitro . Without causing cytotoxicity , WKRG exerted a dose-dependent inhibitory effect on the invasion and motility , but not adhesion , of highly metastatic SK-Hep1 cells . Zymography analyses revealed significant downregulating effects on P08253 , P14780 , and uPA activities in SK-Hep1 cells . Western blot analyses also showed that WKRG treatment caused dose-dependent decreases in P08253 and P14780 protein expressions . Moreover , WKRG increased the levels of P01033 , P16035 , and P05121 . The present study not only demonstrated that invasion and motility of cancer cells were inhibited by WKRG , but also indicated that such effects were likely associated with the decrease in P08253 /-9 and uPA expressions of SK-Hep1 cells . P45983 determines the oncogenic or tumor-suppressive activity of the integrin-linked kinase in human rhabdomyosarcoma . Although most reports describe the protein kinase integrin-linked kinase ( Q13418 ) as a proto-oncogene , occasional studies detail opposing functions in the regulation of normal and transformed cell proliferation , differentiation , and apoptosis . Here , we demonstrated that Q13418 functions as an oncogene in the highly aggressive pediatric sarcoma alveolar rhabdomyosarcoma ( Q9ULH0 ) and as a tumor suppressor in the related embryonal rhabdomyosarcoma ( ERMS ) . These opposing functions hinge on signaling through a noncanonical Q13418 target , P45983 , to the proto-oncogene c-Jun . RNAi-mediated depletion of Q13418 induced activation of JNK and its target , c-Jun , resulting in growth of ERMS cells , whereas in Q9ULH0 cells , it led to loss of JNK/c-Jun signaling and suppression of growth both in vitro and in vivo . Ectopic expression of the fusion gene characteristic of Q9ULH0 ( paired box 3-forkhead homolog in rhabdomyosarcoma [ P23760 - Q12778 ] ) in ERMS cells was sufficient to convert them to an Q9ULH0 signaling phenotype and render Q13418 activity oncogenic . Furthermore , restoration of P45983 in Q9ULH0 reestablished a tumor-suppressive function for Q13418 . These findings indicate what we believe to be a novel effector pathway regulated by Q13418 , provide a mechanism for interconversion of oncogenic and tumor-suppressor functions of a single regulatory protein based on the genetic background of the tumor cells , and suggest a rationale for tailored therapy of rhabdomyosarcoma based on the different activities of Q13418 . Significant association of urokinase plasminogen activator Pro141Leu with serum lipid profiles in a Japanese population . DB00013 plasminogen activator ( uPA ) plays important physiological and pathological roles in fibrinolysis , cancer metastasis , and atherosclerosis . One study suggested that uPA also has a major role in cholesterol biosynthesis in humans via its receptor Q03405 . Thus , we investigated the associations of functional uPA polymorphism ( plasminogen activator , urokinase ; P00749 Pro141Leu , rs2227564 ) with serum lipid profiles in a Japanese cohort . The study included 5152 participants ( 1465 male , 3687 female ; age range , 35-69 years ) of the Daiko Study , a part of the Japan Multi-Institutional Collaborative Cohort Study ( J-MICC Study ) . Subjects were enrolled at the Daiko Medical Center from June 2008 to May 2010 . Low-density lipoprotein cholesterol ( LDL-C ) and non-HDL-C ( subtraction of high-density lipoprotein cholesterol from total cholesterol ) in fasting blood of participants were each classified into two groups , < or ≥ 140 mg/dL , and < or ≥ 170 mg/dL , respectively . Genotype frequencies of P00749 Pro141Leu ( rs2227564 ) were 59.1 % for ProPro , 35.6 % for ProLeu , and 5.3 % for LeuLeu , and were in Hardy-Weinberg equilibrium ( p=0.789 ) . The allele frequencies were 0.769 for Pro and 0.231 for DB00149 . The multivariate-adjusted odd ratios ( ORs ) and 95 % confidence intervals ( CIs ) for high LDL-C and non-HDL-C were 1.11 ( 95 % CI ; 1.00-1.23 ) and 1.16 ( 95 % CI ; 1.03-1.30 ) for those with DB00149 allele relative to ProPro . This study suggested that P00749 Pro141Leu ( rs2227564 ) is significantly associated with serum lipid levels in a Japanese population . Changes in the phenotype of polymorphic plasma proteins after liver transplantation - new data and medico-legal consequences . The genetically inherited polymorphic plasma protein types have always been considered stable for lifetime in humans . Most of these proteins are synthetised in the liver . Phenotypes for 14 plasma proteins in donors and recipients of liver transplants prior to and after transplantation were determined in 15 patients who had undergone liver transplantation at the university hospitals Charité and Rudolf Virchow in Berlin . The plasma proteins investigated were HP , TF , GC , PI , P02763 , ITI , A2HS , P00747 , FXIIIB , BF , P01024 , P13671 , Q99618 and FH . Evidence was provided of irreversible change from the recipient type to the donor type in at least one patient for all the systems investigated . This is the first time such data have been obtained for ITI , A2HS , Q99618 and FH . These results clearly support the point that the dogma of life-long stability of genetically determined protein phenotypes is merely of limited validity . Against the background of good long-term results of liver transplantation , there are consequences for the practice of legal medicine in the particular context of certification of parentage , identification and stain analysis . Protein Tyrosine Phosphatase 1B ( P18031 ) deficiency accelerates hepatic regeneration in mice . Protein tyrosine phosphatase 1B ( P18031 ) is a key regulator of metabolism and cell growth by its ability to dephosphorylate tyrosine kinase receptors and modulate the intensity of their signaling cascades . Because liver regeneration involves tyrosine phosphorylation-mediated signaling , we investigated the role of P18031 in this process by performing partial hepatectomy in wild-type ( P18031 (+/+) ) and P18031 -deficient ( P18031 (-/-) ) mice . The expression of P12004 and cyclins D1 and E ( cell proliferation markers ) was enhanced in P18031 (-/-) regenerating livers , in parallel with 5'-bromo-2'-deoxyuridine incorporation . Phosphorylation of P45983 /2 and P40763 , early triggers of hepatic regeneration in response to P01375 -α and P05231 , was accelerated in P18031 (-/-) mice compared with P18031 (+/+) mice . These phosphorylations were increased in P18031 (-/-) hepatocytes or by silencing P18031 in wild-type cells and decreased further after the addition of recombinant P18031 . Enhanced P01133 - and P08581 -mediated signaling was observed in regenerating livers lacking P18031 and in P01133 - or P14210 -stimulated P18031 (-/-) hepatocytes . Moreover , P18031 (-/-) mice displayed a more rapid increase in intrahepatic lipid accumulation than P18031 (+/+) control mice . Late responses to partial hepatectomy revealed additional divergences because stress-mediated signaling was attenuated at 24 to 96 hours in P18031 (-/-) mice compared with P18031 (+/+) mice . Finally , P18031 deficiency also improves hepatic regeneration in mice fed a high-fat diet . These results suggest that pharmacological inhibition of P18031 would improve liver regeneration in patients with acute or chronic liver injury . Mediators of chronic inflammation in polycystic ovarian syndrome . Polycystic ovarian syndrome ( PCOS ) is an endocrine disorder affecting 5-10 % of reproductive-age women . Hyperandrogenemia , which characterizes the syndrome , stimulates the maturation of adipocytes and favors central obesity . The linking hub between obesity and other metabolic manifestations of the syndrome seems to be chronic low-grade inflammation . We discuss the most reliable current data regarding the role of inflammatory mediators in PCOS , with particular focus on the genetic mechanisms implicated . P02741 levels are 96 % higher in PCOS patients than in healthy controls . Patients with the -308A polymorphism of the tumor necrosis factor-α gene have elevated androgens in comparison with carriers of the -308G . Interleukin 18 ( Q14116 ) is elevated in lean patients , with a further rise in the presence of obesity and insulin resistance . Polymorphisms of the IL-1a , IL-1b and P05231 genes have also been associated with PCOS . P00747 activator inhibitor-1 levels are positively associated with the syndrome , and carriers of the 4G allele of the 4G/5G polymorphism are at risk of developing PCOS . Other mediators discussed include adhesion molecules , osteoprotegerin , asymmetric dimethylarginine , homocysteine and advanced glycation end-products . The elucidation of the pathogenetic mechanisms implicated in PCOS and their connection with low-grade inflammation may in the future offer the opportunity for the formulation of novel therapeutic strategies and individualized therapy for these patients . P00747 activation by airway smooth muscle is regulated by type I collagen . Plasmin , the activated protease product of plasminogen , is involved in collagen remodeling , and is strongly implicated in asthma pathophysiology by recent genome-wide association studies . This study examines plasminogen " activation " by airway smooth muscle cells , and its regulation in a fibrotic environment created by culture on type I collagen and incubation with transforming growth factor ( TGF ) -β . DB00013 plasminogen activator ( uPA ) activity was detected in the supernatants of human airway smooth muscle cell cultures maintained in serum-free conditions . Incubation with plasminogen ( 1.5-50.0 μg/ml , 24 h ) increased plasmin activity in a concentration-dependent manner ( P < 0.001 ) . uPA activity was higher in cultures maintained on fibrillar type I collagen substrata than in those on plastic , as was plasmin activity after incubation with plasminogen ( 20 μg/ml ) . Pretreatment with TGF-β ( 100 pM ) for 18 hours inhibited plasminogen activation by airway smooth muscle cells maintained on plastic , but not on collagen . TGF-β stimulated an increase in the level of uPA mRNA in airway smooth muscle cells grown on collagen , but not on plastic . Reducing the levels of β1-integrin collagen receptor , using interference RNA , attenuated plasmin formation by airway smooth muscle cells grown on collagen , and restored the inhibitory effect of TGF-β . This study shows that airway smooth muscle activation of plasminogen by uPA is accelerated in a collagen-rich environment in which the inhibitory effect of TGF-β is attenuated in association with greater uPA expression induced via β1-integrin signaling . These findings suggest that the plasminogen-activation system involving uPA has the potential to contribute to airway wall remodeling in asthma . DB00169 modulation of plasminogen activator inhibitor type-1 in human breast carcinomas under organ culture . DB00013 plasminogen activator ( uPA ) , its cell-bound receptor ( Q03405 ) and its main inhibitor plasminogen activator type 1 ( P05121 ) are present primarily in stromal cells in invasive breast carcinoma . The purpose of this study was to investigate the regulation by 1,25 dihydroxyvitamin-D3 ( VD3 ) of these invasion-associated markers expressed in breast cancer tumors under organ culture , which preserves the interacting network of tumor and stromal cells . Breast carcinoma slices ( 30 cases ) , obtained using the Krumdieck tissue slicer , cultured for 48 h in the presence or absence of 100 nM vitamin D3 , were embedded in formalin-fixed paraffin . uPA , Q03405 , P05121 and VD3 receptor ( P11473 ) were analyzed by immunohistochemistry , and their expression , detected in tumor cells and fibroblasts of the specimens , was not statistically changed by culture conditions . The proportion of cases expressing uPA , Q03405 and P05121 was not affected by VD3 in epithelial cells , but the fraction of cases displaying strong P05121 reactivity in fibroblasts was reduced ( P=0.016 ) compared with control slices . Fibroblasts isolated from invasive ductal carcinomas and from normal breast tissues expressed higher P11473 mRNA levels than epithelial cells . In cultured tumor fibroblasts , P05121 immunostaining and mRNA levels were reduced by VD3-limiting fibroblast contribution to invasion . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . DB00013 mediates fibrinolysis in the pulmonary microvasculature . The role of urokinase-type plasminogen activator ( uPA ) and its receptor ( Q03405 ) in fibrinolysis remains unsettled . The contribution of uPA may depend on the vascular location , the physical properties of the clot , and its impact on tissue function . To study the contribution of urokinase within the pulmonary microvasculature , a model of pulmonary microembolism in the mouse was developed . Iodine 125 ( (125)I ) -labeled fibrin microparticles injected intravenously through the tail vein lodged preferentially in the lung , distributing homogeneously throughout the lobes . Clearance of (125)I-microemboli in wild type mice was rapid and essentially complete by 5 hours . In contrast , uPA(-/-) and tissue-type plasminogen activator tPA(-/-) mice , but not Q03405 (-/-) mice , showed a marked impairment in pulmonary fibrinolysis throughout the experimental period . The phenotype in the uPA(-/-) mouse was rescued completely by infusion of single chain uPA ( scuPA ) . The increment in clot lysis was 4-fold greater in uPA(-/-) mice infused with the same concentration of scuPA complexed with soluble recombinant Q03405 . These data indicate that uPA contributes to endogenous fibrinolysis in the pulmonary vasculature to the same extent as tPA in this model system . Binding of scuPA to its receptor promotes fibrinolytic activity in vivo as well as in vitro . The physical properties of fibrin clots , including size , age , and cellular composition , as well as heterogeneity in endothelial cell function , may modify the participation of uPA in endogenous fibrinolysis . ( Blood. 2000;96:1820-1826 ) Eupolyphaga sinensis walker displays inhibition on hepatocellular carcinoma through regulating cell growth and metastasis signaling . Tumor growth and metastasis are responsible for most cancer patients ' deaths . Here , we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis . Compared with proliferation , colony formation , transwell assay and transplantable tumor in nude mouse in vitro and vivo , eupolyphaga sinensis walker extract ( ESWE ) showed good inhibition on the SMMC-7721 cell growth and metastasis . Using genome-wide microarray analysis , we found the down-regulated growth and metastasis factors , and selected down-regulated genes were confirmed by real-time PCR . Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE . Moreover , our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ , Akt , m-TOR , Erk1/2 , MEK-2 , Raf and JNK-1 , but also increasing cyclin D1 protein levels and decreasing amount of cyclin E , cyclin B1 and cdc2 of the cycle proteins . At the same time , ESWE reduced P08253 , P14780 and P61073 , P00747 , NFκB and P04637 activities . Overall , our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC , AKT , MAPK signaling and related metastasis signaling , having potential in cancer therapy . [ Index of fibrinolysis with new fibrin plate ( author 's transl ) ] . P00747 -free fibrin plate ( fP ) which was made from treated commerical bovine fibrinogen with DB00123 -Sepharose was developed in our laboratory . This new fibrin plate showed the following specificities . a ) This new fibrin plate did not show any lysis with high amount of streptokinase and DB00013 ( 10,000 u/ml and 500 u/ml ) . b ) The concentrations of its substrate was the same as standard plate ( SP ) and its substrate was not denatured compared with heated plate (HP). c ) The activity of plasmin can be measured quantitatively on fP and linear correlation between plasmin units and lysis area was showen . d ) This procedure of new fibrin plate was easy and simple and could be applicable to the materials of other species , i.e. , human , rabbit and porcine . With the use of two kinds of bovine fibrin plate ( SP and fP ) , activation of fibrinolysis of human plasma , euglobulin and plasminogen induced by SK and UK was investigated and each correlation ship between sample and activator was studied statistically . From these results , " Index of fibrinolysis " meaning of fibrinolytic components such as plasmin , plasminogen , activator , proactivator , anti-activator and anti-plasmin were indicated . Indeed , these index of fibrinolysis were calculated from the lysis area of plasma+SK , Eug.+SK and Eug.+UK by each formula and index obtained from some physiological and pathological condition showed us many new information about fibrinolysis . DB00013 plasminogen activator receptor promotes macrophage infiltration into the vascular wall of ApoE deficient mice . The urokinase plasminogen activator receptor ( Q03405 ) regulates macrophage adhesion and migration by binding directly to matrix proteins and signaling through integrin complexes . In this study , we examined the role of Q03405 on macrophage infiltration into the vascular wall . Stable murine macrophage ( Raw264.7 ) cell lines expressing high levels of human Q03405 , human urokinase plasminogen activator ( uPA ) , or both were established using expression vectors driven by the human P34810 promoter . Stimulation with human uPA specifically induced phosphorylation of early response regulated kinase ( P29323 ) in cells expressing human Q03405 but not in sham transfected cells . The human Q03405 expressing Raw264.7 cells showed increased adhesion to both human uPA and vitronectin ( Vn ) . Raw264.7 cells expressing human Q03405 or both human Q03405 and uPA , but not uPA alone , were detected in the aortic wall of ApoE(-/-) mice , and no cells were detected in that of age-matched C57BL/6J mice after intravenous infusion of the cells . Blocking of Mac-1/ P05362 interaction by anti-alphaM antibody ( M1/70 ) significantly reduced the infiltration of huPAR-expressing Raw264.1 cells into aorta of ApoE(-/-) mice . Treatment of C57BL/6J mice with angiotensin II resulted in infiltration of Raw264.7 cells expressing human Q03405 . These data demonstrate that Q03405 plays a key role in promoting macrophage infiltration into the arterial wall of ApoE(-/-) mice . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Co-expression of Q03405 and P61073 promotes tumor growth and metastasis in small cell lung cancer . P00749 receptor ( Q03405 ) and C-X-C-chemokine receptor-4 ( P61073 ) are considered as key molecules in invasion and metastasis of several cancers via extracellular matrix degeneration and assist tumor metastasis to specific sites by chemotaxis . However , the combined effect of Q03405 and P61073 on small cell lung cancer ( SCLC ) , the most aggressive type of lung cancer , is not clear . In this study , we detected the expression of Q03405 and P61073 in SCLC tissue samples ( n = 50 ) by immunohistochemistry . The tumors with high expression of both Q03405 and P61073 ( 12/50 ) had larger size , higher lymph node ( LN ) metastasis and worse prognosis of patients than those with low expression of Q03405 and P61073 ( 38/50 ) ( P < 0.05 ) . We further identified and isolated the both Q03405 and P61073 positive expression subpopulation cells ( Q03405 (+) P61073 (+) cells ) from the SCLC cell line H446 by flow cytometry . The Q03405 (+) P61073 (+) cancer cells showed a higher invasive and migrating capacity in the transwell and wound healing assays compared with other subpopulation cells ( P < 0.05 ) . Q03405 (+) P61073 (+) cells injected subcutaneously in nude mice markedly increased tumor growth and induced lung metastasis , while other subpopulation cells did not . In conclusion , these data suggest that Q03405 and P61073 co-expression predicts worse prognosis of SCLC patients . Q03405 (+) P61073 (+) cells promote the tumor growth and play a potential role in metastasis of SCLC . Components of the plasminogen activation system in uveal melanoma -- a clinico-pathological study . In tumour development , proteases such as plasminogen activators ( PAs ) play a role in degradation of the extracellular matrix and other tissue barriers . Recently , we demonstrated that plasminogen activators , their inhibitors , and urokinase receptor emerge in late stages of cutaneous melanocytic tumour progression . In this study we investigated the expression and distribution of the various components of the PA system and the presence of PA enzyme activity in 45 freshly frozen primary uveal melanoma with known follow-up ( 14 spindle and 31 non-spindle type ) and in metastases ( n = 5 ) . Tissue-type PA ( t-PA ) was found in endothelium of blood vessels and in tumour cells in almost all lesions , and was markedly present at the invasive front ( towards the sclera and Bruch 's membrane ) , but no correlation with tumour-related death could be established . DB00013 PA ( u-PA ) was expressed focally , by only five non-spindle cell melanomas but in all metastases . u-PA expression correlated with occurrence of metastasis . u-PA receptor ( u-PAR ) was present in one-third of all the tumours examined . P00747 activator inhibitors ( P05121 and P05120 ) were found only focally in approximately 10 per cent of the lesions . Staining of t-PA , u-PA , and P05121 was observed in all the metastases . We conclude that in uveal melanoma , u-PA expression may be associated with metastatic disease and accordingly with a poor prognosis . Further research on a larger group of tumours with known follow-up is needed to establish whether u-PA positivity is of additional prognostic value in uveal melanoma . Modulation of the gene network connected to interferon-gamma in liver regeneration from oval cells . Suppression subtractive hybridization was used to clone genes associated with proliferation of oval cells in rat liver regenerating after a 70 % partial hepatectomy combined with the feeding of 2-acetylaminofluorene . A subset of the identified genes comprised interferon-gamma receptor alpha subunit ( IFN-gammaRalpha ) , gp91phox , interleukin-1beta ( IL-1beta ) , lymphocyte function-associated molecule-1alpha ( LFA-1 ) , eukaryotic initiation factor-2-associated 67-kd protein ( eIF-2-associated 67-kd protein ) , and alpha-fetoprotein , which constitute part of the cellular program modulated by P01579 . Therefore , expression analysis performed by Northern blotting and immunohistochemistry were extended to include P01579 , the P01579 receptor beta subunit ( IFN-gammaRbeta ) , three secondary response genes induced by interaction of P01579 with P01579 receptor complexes , ie , IL-1beta-converting enzyme ( ICE ) , intercellular adhesion molecule-1 ( P05362 ) , and urokinase-type plasminogen activator receptor ( Q03405 ) , and a cytokine inducing P01579 expression , ie , interleukin-18 ( Q14116 ) . The Northern blot analysis showed that all examined genes were modulated when progenitor-like oval cells were activated and recruited for liver regeneration . Immunohistochemistry localized the subunits of the P01579 receptor complex , IFN-gammaRalpha and IFN-gammaRbeta , the secondary response genes Q03405 and P05362 , the Q14116 Q14116 , and ICE to the ductular structures of oval cells . In contrast , during liver regeneration after a 70 % partial hepatectomy , only modulation of IL-1beta and ICE was observed . Our results , therefore , indicate that P01579 -mediated events may be particularly important when cells in the bile ductules must respond to liver damage by production of ductular oval cells . DB00013 -type P00747 Activator ( uPA ) is Inhibited with QLT0267 a Small Molecule Targeting Integrin-linked Kinase ( Q13418 ) . P00749 ( uPA ) is associated with cancer recurrence where the most evidence comes from studies in breast cancer . According to the European Organization for Research and Treatment of Cancer , uPA is considered one of the most prominent biomarkers for cancer recurrence and therefore new agents are needed to inhibit it . Whether uPA is also expressed in pediatric cancers is yet unknown . If it is then uPA inhibitors might also help children with recurrent cancers . In this study , we addressed whether the integrin-linked kinase inhibitor ( Q13418 ) , QLT0267 , could suppress uPA . We previously showed that uPA expression is maximally inhibited when both the Akt and Q96HU1 kinase pathways were blocked which we anticipated can be achieved via QLT0267 . In MDA-MB-231 breast cancer cells , QLT0267 blocked signaling through Akt and Q96HU1 kinase with a correlative decrease in uPA protein and mRNA , which corresponded to an inhibition of c-Jun phosphorylation . Consistent with these findings , cellular invasion was inhibited with either QLT0267 or with small interfering RNA against Q13418 . We then questioned whether uPA was commonly expressed in childhood sarcomas and if QLT0267 might be effective in this setting . We determined for the first time that uPA was highly expressed in rhabdomyosarcomas ( RMS ) , but not Ewings sarcomas by screening cell lines ( n = 31 ) and patient samples ( n = 200 ) using Affymetrix microarrays . In alveolar RMS ( Q9ULH0 ) cell lines , QLT0267 blocked cell signaling , uPA production , invasion and ultimately survival . We concluded that QLT0267 blocks the production of uPA providing a new target for the management of recurrent cancers . Molecular imaging of pancreatic cancer in an animal model using targeted multifunctional nanoparticles . BACKGROUND & AIMS : Identification of a ligand/receptor system that enables functionalized nanoparticles to efficiently target pancreatic cancer holds great promise for the development of novel approaches for the detection and treatment of pancreatic cancer . DB00013 plasminogen activator receptor ( Q03405 ) , a cellular receptor that is highly expressed in pancreatic cancer and tumor stromal cells , is an excellent surface molecule for receptor-targeted imaging of pancreatic cancer using multifunctional nanoparticles . METHODS : The Q03405 -targeted dual-modality molecular imaging nanoparticle probe is designed and prepared by conjugating a near-infrared dye-labeled amino-terminal fragment of the receptor binding domain of urokinase plasminogen activator to the surface of functionalized magnetic iron oxide nanoparticles . RESULTS : We have shown that the systemic delivery of Q03405 -targeted nanoparticles leads to their selective accumulation within tumors of orthotopically xenografted human pancreatic cancer in nude mice . The Q03405 -targeted nanoparticle probe binds to and is subsequently internalized by Q03405 -expressing tumor cells and tumor-associated stromal cells , which facilitates the intratumoral distribution of the nanoparticles and increases the amount and retention of the nanoparticles in a tumor mass . Imaging properties of the nanoparticles enable in vivo optical and magnetic resonance imaging of Q03405 -elevated pancreatic cancer lesions . CONCLUSIONS : Targeting Q03405 using biodegradable multifunctional nanoparticles allows for the selective delivery of the nanoparticles into primary and metastatic pancreatic cancer lesions . This novel receptor-targeted nanoparticle is a potential molecular imaging agent for the detection of pancreatic cancer . Semi-quantitation of urokinase plasminogen activator and its receptor in breast carcinomas by immunocytochemistry . DB00013 plasminogen activator ( uPA ) is a serine protease involved in cancer invasion and metastasis . uPA acts in vivo by binding to a membrane receptor known as Q03405 . In this study , uPA and Q03405 levels were semiquantitated by immunocytochemistry in 36 primary breast carcinomas . Using monoclonal antibody HD-UK 1 , uPA was detected both in stromal and in malignant cells . However , the predominant location was in the stromal cells . Using double-staining , cells containing uPA were also found to coexpress either cytokeratin ( an epithelial cell marker ) or more frequently KP1 ( a macrophage/monocyte cell marker ) . With monoclonal antibody HD- Q03405 13.1 , Q03405 was localized principally to spindle- or macrophage-like stromal cells , especially when these cells surrounded invasive breast cancer . In contrast , Q03405 was only rarely detected in cancer cells and was not detected in normal epithelia surrounding tumour or in areas of adenosis . uPA levels in both stromal and epithelial cells were significantly correlated with those for Q03405 . We conclude that both uPA and its receptor are mostly present in stromal cells in invasive breast carcinomas . These results suggest that stromal cells collaborate with malignant cells to mediate metastasis . Differential transmission of Q13233 morphogenetic signals by P45983 and P45984 . P45983 and P45984 are two ubiquitously expressed isoforms that exert redundant roles in many physiological processes , but the extent of their relative contributions to these processes has not been well characterized . We show that both JNK isoforms transmit Q13233 ( Q13233 ) -mediated morphogenetic signals during mouse embryonic eyelid closure . However , P45983 and P45984 are not synonymous , because Q13233 is haploinsufficient for normal eyelid closure in Jnk1-null mice , but is haplosufficient in Jnk2-null mice . In the Mekk1 heterozygous background , a more efficient phosphorylation of P45983 than P45984 leads to differential downstream reactions , such as c-Jun phosphorylation and P05121 expression in the developing eyelid epithelium . Differences in efficiency of phosphorylation are attributed to P45983 Gly177 and Ser179 -- residues that are absent in P45984 -- which promote a less ordered structural conformation . This leads to more favorable JNK phosphorylation by activin B morphogenetic signals mediated by the Q13233 - P45985 pathway . Interestingly , Mekk1-Jnk1-Jnk2 triple hemizygotes display a partial eye-open phenotype at birth , suggesting that all three genes dose-dependently contribute to morphogenetic eyelid closure . We propose that a Q13233 - P45983 /2 axis governs the JNK activation levels to control downstream transcriptional events and eyelid morphogenesis and that reduction of upstream Q13233 signals uncovers analogous but differential roles of P45983 and P45984 in a biological process . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . The pivotal role of astrocytes in an in vitro stroke model of the blood-brain barrier . Stabilization of the blood-brain barrier during and after stroke can lead to less adverse outcome . For elucidation of underlying mechanisms and development of novel therapeutic strategies validated in vitro disease models of the blood-brain barrier could be very helpful . To mimic in vitro stroke conditions we have established a blood-brain barrier in vitro model based on mouse cell line cerebEND and applied oxygen/glucose deprivation ( OGD ) . The role of astrocytes in this disease model was investigated by using cell line P13671 . Transwell studies pointed out that addition of astrocytes during OGD increased the barrier damage significantly in comparison to the endothelial monoculture shown by changes of transendothelial electrical resistance as well as fluorescein permeability data . Analysis on mRNA and protein levels by qPCR , western blotting and immunofluorescence microscopy of tight junction molecules claudin-3,-5,-12 , occludin and ZO-1 revealed that their regulation and localisation is associated with the functional barrier breakdown . Furthermore , soluble factors of astrocytes , OGD and their combination were able to induce changes of functionality and expression of ABC-transporters Abcb1a ( P-gp ) , Abcg2 ( bcrp ) , and Abcc4 ( mrp4 ) . Moreover , the expression of proteases ( matrixmetalloproteinases P08253 , P08254 , P14780 , and t-PA ) as well as of their endogenous inhibitors ( P01033 , P35625 , P05121 ) was altered by astrocyte factors and OGD which resulted in significant changes of total MMP and t-PA activity . Morphological rearrangements induced by OGD and treatment with astrocyte factors were confirmed at a nanometer scale using atomic force microscopy . In conclusion , astrocytes play a major role in blood-brain barrier breakdown during OGD in vitro . DB00013 , tissue-type plasminogen activator and plasminogen activator inhibitor-1 expression in severely stenosed and occluded vein grafts with thrombosis . Intimal hyperplasia and subsequent thrombotic occlusions limit the success of vascular reconstructive procedures . P00747 activation in situ may be an important factor affecting re-stenosis of the graft . Tissue specimens from eight patients with failing or failed infra-inguinal vein bypasses and three specimens from normal veins were harvested to study urokinase-type plasminogen activator ( u-PA ) , tissue-type plasminogen activator ( t-PA ) and plasminogen activator inhibitor-1 ( P05121 ) by in situ hybridization and immunohistochemistry . The possible presence of thrombi was monitored by platelet and fibrin-specific stainings . In occluded grafts , platelet endothelial cell adhesion molecule ( P16284 ) antibody stained the thrombi but not the endothelial area , indicating the absence of endothelium . Platelet glycoprotein ( GP ) IIb/IIIa co-localized with P16284 and , furthermore , GP IIb/IIIa staining was positive on the vein walls with thrombi and to some extent in the grafts without thrombi . P05121 and u-PA were uniformly upregulated in intimal thickening in grafts without thrombus . In organized thrombi , enhanced u-PA , t-PA and P05121 reactivity was detected in the ingrowing subendothelium . In non-occluded grafts with small thrombi , u-PA expression was enriched beneath microthrombi co-localizing with the graft wall injury , while P05121 was scattered in the (sub)endothelium . We conclude that fibrinolytic system is upregulated at sites of graft stenosis , and local proteolytic degradation of the graft wall associates with thrombus formation . The plasminogen-activating system in hepatic stellate cells . DB00013 plasminogen activator ( uPA ) generates plasmin , a process inhibited by plasminogen-activator inhibitor ( P05121 ) -1 and localized to the cell surface by binding of uPA to a specific receptor . Plasmin degrades extracellular matrix ( Q13201 ) both directly and by activation of matrix metalloproteinases ( MMPs ) . Because stellate cells play a central role in the pathogenesis of liver fibrosis both via production of Q13201 proteins and through secretion of MMPs , their contribution to plasmin generation was assessed . Stellate cells were prepared from rat liver and cultured on plastic . Northern analysis showed cellular expression of messenger RNA ( mRNA ) for P05121 , uPA , and uPA receptor . Zymography/reverse zymography identified cell-surface-associated uPA activity and uPA and P05121 in culture media . Net uPA activity in culture media was maximal after 7 days in culture and then declined , whereas P05121 antigen levels remained consistently elevated between 7 and 21 days in culture . Stellate cell-mediated plasmin generation was also seen in in vitro cultures supplemented with plasminogen . Because hepatic stellate cells ( HSCs ) contain retinoids and release them on activation , the effect of retinoic acid on the plasminogen-activating system was also assessed . Treatment of cultured HSCs with retinoic acid ( 1 micromol/L ) increased uPA secretion 2.6-fold but did not alter P05121 . We conclude that stellate cells synthesize key components of the plasminogen-activating system and generate plasmin and therefore have the ability to regulate MMP activation . Upregulation of uPA synthesis by retinoic acid may have implications in matrix remodeling in sites of stellate cell activation in which high concentrations of retinoids may be achieved . DB00013 expression and binding activity associated with the transforming growth factor beta1-induced migratory and invasive phenotype of mouse epidermal keratinocytes . Transforming growth factor beta1(TGF-beta1) is a stimulator of malignant progression in mouse skin carcinogenesis . TGF-beta1 exerts a differential effect on cultured nontumorigenic ( MCA3D cell line ) and transformed ( PDV cell line ) keratinocytes . Whereas MCA3D cells are growth arrested and committed to die in the presence of the factor , it induces a reversible epithelial-fibroblastic conversion in PDV cells . This conversion is associated in vivo with a squamous-spindle cell carcinoma transition . Here we have investigated the role of urokinase ( uPA ) during malignant progression of transformed epidermal keratinocytes . We show that the levels of uPA expression/secretion , and the uPA binding activity to the cell surface , correlate with the invasive and malignant potentials of mouse epidermal cell lines . TGF-beta1 enhanced uPA production , the number of uPA cell surface binding sites , and the expression of the plasminogen activator inhibitor P05121 , in transformed PDV cells , but had no major effect on nontumorigenic MCA3D keratinocytes . Increased uPA production depended on the presence of the factor in the culture medium and occurred concomitantly to the stimulation of the migratory and invasive abilities of PDV cells . Synthetic peptides containing the amino terminal sequence of the mature mouse uPA inhibited the binding of uPA to the cell surface and decreased TGF-beta1-induced cell motility and invasiveness . These results demonstrate that the uPA system mediates at least part of the migratory and invasive phenotype induced by TGF-beta1 in transformed keratinocytes , and suggest a role for uPA on the changes that lead to the appearance of spindle carcinomas . DB00013 plasminogen activator induces angiogenesis and tumor vessel invasion in breast cancer . DB00013 plasminogen activator ( uPA ) is a proteolytic enzyme implicated in cancer invasion and tumor progression . DB00013 PA and its inhibitor ( P05121 ) appear to be new and independent prognostic markers in breast cancer . To investigate how uPA- and P05121 -levels correlate with angiogenesis and tumor vessel invasion , we counted microvessels and their tumor invasion and determined the uPA- and P05121 levels in 42 primary invasive breast carcinomas . 20 Patients had no lymph node metastasis at the time of surgery , while 22 patients had positive nodes . Using light microscopy , we highlighted the vessels by staining their endothelial cells immunocytochemically for CD31 and Factor VIII . After gaining tumor tissue extracts , we determined the uPA- and P05121 -levels by ELISA . A positive correlation between microvessel density , angioinvasion and uPA- and P05121 -levels was found . We speculate that high uPA levels may induce tumor neovascularisation , angioinvasion and may cause tumor progression and metastasis . The degradation of the vessel wall by uPA causes a leak . This wall defect may , on the one hand , be the stimulus for endothelial cell proliferation and formation of new blood vessels and , on the other hand , it may be the place of tumor cell entry . DB00013 receptor , P03956 and P14780 are markers to differentiate prognosis , adenoma and carcinoma in thyroid malignancies . The identification of high-risk patients with thyroid cancer and the preoperative differentiation between follicular adenoma and carcinoma remain clinically challenging . Our study was conducted to analyze whether the quantification of matrix metalloproteinases ( MMPs ) and urokinase-type plasminogen activator receptor ( u-PAR ) and transcription factor binding to the u-PAR promoter improve prognostic predictability and differential diagnosis of thyroid tumors . Tumor/normal tissue was collected from 69 prospectively followed patients with thyroid carcinomas ( papillary , medullary , follicular and anaplastic , PTC , P04629 , FTC and ATC ) or follicular adenomas . Q03405 , P03956 , P09237 and P14780 amounts were determined by ELISA , and transcription factor binding was determined by electrophoretic mobility shift assay . Binding of transcription factors to the u-PAR promoter was observed , but not associated with u-PAR expression . Carcinomas except P04629 expressed significantly more u-PAR/MMPs than adenomas/normal tissues , this being associated with advanced pT- or M-stages . P03956 and P14780 were significantly higher in follicular carcinomas than in adenomas . In carcinomas , high u-PAR-gene expression correlated significantly with high P14780 , the latter being associated with P09237 in normal tissues . Poor survival in differentiated tumors was associated in trend ( p = 0.07 ) ; poor survival of all patients ( p = 0.043 ) and especially of patients with carcinomas of follicular origin ( including ATC ) , but not medullary carcinomas , were significantly associated with high u-PAR-protein ( p = 0.015 ) . Quantification of u-PAR is of prognostic relevance in thyroid carcinomas of non-c-cell origin , and u-PAR in part may be regulated nontranscriptionally in thyroid cancers . This is the first study to suggest P03956 /-9 as significant differentiation markers between follicular adenoma and follicular carcinoma . DB00013 and plasminogen activator-inhibitor ( P05121 ) status in primary ovarian carcinomas and ovarian metastases compared to benign ovarian tumors as a function of histopathological parameters . Ninety-eight patients with histologically confirmed ovarian tumors ( 77 primary ovarian carcinomas of stages T1 to DB00279 according to the postoperative histopathological classification pTNM classification , 14 ovarian metastases of various origins and seven benign ovarian tumors ) were investigated with regard to the concentration of urokinase-type plasminogen activator ( uPA ) and plasminogen activator inhibitor ( P05121 ) in membrane extracts of tumors . The results were correlated with the clinical course and with histopathological findings . With more advanced stage of primary ovarian carcinomas , there was a highly significant rise in the membrane concentrations of both uPA and P05121 . However , increasing dedifferentiation of the tumors correlated only with uPA , but not with P05121 . There was no correlation between the number of steroid receptors for estradiol and progesterone and the content of uPA or P05121 in the primary ovarian carcinomas . In the 14 ovarian metastases of different origins incluced in the study , the contents of uPA and P05121 were comparable to those of primary ovarian carcinomas . Compared with the malignant ovarian tumors , the median uPA and P05121 concentrations in the membrane fraction were 2.5-6 fold lower ( highly significant ) in the group of seven benign tumors . A cut-off value of 4.8ng/mg pellet protein for a prognostically favorable ( < 4.8 ) or unfavorable course ( > 4.8 ) could be determined for uPA ( p = 0.0392 ) but not for P05121 on the basis of the Kaplan and Meier survival curves in the malignant primary ovarian carcinomas . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . DB01016 exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803 . DB01016 , a blocker of DB00171 -sensitive potassium ( K( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC-803 cells expressed the K( DB00171 ) channels composed of Kir6.2 and Q09428 subunits . DB01016 induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC-803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion ( O2- ) generation , and caused mitochondrial respiration dysfunction in MGC-803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2-terminal kinase ( JNK ) in MGC-803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 -/- or P45984 -/- MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer . Rac1 modulates acute and subacute genotoxin-induced hepatic stress responses , fibrosis and liver aging . To investigate the importance of the Ras-homologous GTPase Rac1 for the hepatic response to genotoxic insults and liver aging , rac1 was deleted in liver of mice by Mx1-Cre-based recombination . Knockout of rac1 caused complex changes in basal as well as doxorubicin and ionizing radiation-induced mRNA expression of various genotoxic stress response-related genes , including hspa1b , rad51 , wrn and xpc . Rac1 deletion protected the liver from acute toxicity following doxorubicin treatment . Moreover , the level of S139 phosphorylated histone P16104 ( γ P16104 ) , which is indicative of DNA damage , and mRNA expression of pro-inflammatory ( P05231 ) and pro-fibrotic ( P29279 , TGFβ , αSMA ) factors were mitigated in rac1 knockout animals . By contrast , lack of rac1 promoted subacute hepatotoxicity , which was determined 3 weeks after injection of multiple low doses of doxorubicin by assaying the γ P16104 level , mitotic index and pro-fibrotic gene expression . Regarding ionizing radiation , rac1 deficiency had no major effects on DNA damage induction or acute pro-inflammatory and pro-fibrotic stress responses . Mice lacking hepatic rac1 for extended period of time ( 15 months ) revealed increased mRNA expression of fibrosis-related factors ( P29279 , TGFβ , collagen , P03956 ) and fibrotic tissue remodeling . In addition , protein expression of the senescence marker p16 was enhanced in the absence of rac1 . Taken together , the data provide evidence that Rac1 is required for doxorubicin-induced DNA damage induction . It is also involved in both the acute and delayed inflammatory and fibrotic stress response in the liver following doxorubicin , but not ionizing radiation , treatment and , furthermore , protects against endogenous liver aging . DB00013 type plasminogen activator and its receptor regulate the invasive potential of gastric cancer cell lines . To assess the role of urokinase-type plasminogen activator ( uPA ) and uPA receptor ( Q03405 ) on the invasive potential of cancer cells , in vitro experiments were performed using two human gastric cancer cell lines , NUGC-3 and MKN-28 . NUGC-3 cells secreted a higher level of uPA than MKN-28 cells , while the Q03405 expression of NUGC-3 cells was lower than that of MKN-28 cells . Both cancer cell lines expressed DB00134 protein and did not express hepatocyte growth factor ( P14210 ) . In Matrigel invasion assay , MKN-28 cells demonstrated significantly lower invasion index than NUGC-3 cells . The addition of exogenous uPA significantly increased the invasive activity of MKN-28 cells . The uPA expression in NUGC-3 cells was enhanced by adding conditioned media of fibroblast cells or P14210 . These results suggest that uPA promotes the invasive capacity of the Q03405 -positive cancer cells , and that stromal cells may play an important role in cancer cell invasion by supplying uPA and/or promoting uPA production . DB00013 -deficient and urokinase receptor-deficient mice have impaired neutrophil antimicrobial activation in vitro . Leukocytes express both urokinase-type plasminogen activator ( uPA ) and the urokinase receptor ( Q03405 , CD87 ) . We have shown that neutrophil recruitment to the lung during P. aeruginosa pneumonia is impaired in Q03405 -deficient ( Q03405 -/- ) mice but is normal in uPA-/- mice . However , both uPA-/- mice and Q03405 -/- mice have impaired lung clearance of P. aeruginosa compared with wild-type ( WT ) mice . To determine the role of uPA and Q03405 in antibacterial host defense , we compared neutrophil bacterial-phagocytosis , respiratory burst , and degranulation among uPA-/- , Q03405 -/- , and WT mice . Neutrophil phagocytosis was significantly diminished comparing uPA-/- and Q03405 -/- mice with WT mice at all time points . The generation of superoxide by both uPA-/- and Q03405 -/- neutrophils was about half of that seen in WT neutrophils . Degranulation of azurophilic granules was significantly diminished in uPA-/- neutrophils compared with either Q03405 -/- or WT neutrophils . By contrast , agonist-stimulated release of specific granules was not diminished in either uPA-/- or Q03405 -/- mice compared with WT . We conclude that the uPA/ Q03405 system modulates several of the crucial steps in neutrophil activation that result in bacterial killing and effective innate host defense . DB00013 -dependent human vascular smooth muscle cell adhesion requires selective vitronectin phosphorylation by ectoprotein kinase CK2 . DB00013 ( uPA ) - and urokinase receptor ( Q03405 ) -dependent cell adhesion to the extracellular matrix protein vitronectin ( Vn ) is an important event in wound healing , tissue remodeling , immune response , and cancer . We recently demonstrated that in human vascular smooth muscle cells ( VSMC ) uPA/ Q03405 are functionally associated with the ectoprotein kinase casein kinase-2 ( CK2 ) . We now asked whether CK2 regulates uPA-dependent cell adhesion to Vn , since the latter is a natural CK2 substrate . We found that Vn is indeed selectively phosphorylated by CK2 and that this phosphorylation is uPA-regulated in VSMC . Vn induces release of ecto-CK2 from the cell surface via a process termed as " shedding. " CK2-mediated Vn phosphorylation was decisive for the uPA-dependent VSMC adhesion . Specific inhibition of CK2 completely abolished the uPA-induced cell adhesion to Vn . This effect was specific for cell adhesion to Vn and required participation of both Q03405 and alpha(v)beta(3) integrins as adhesion receptors . CK2 localization at the cell surface was highly dynamic ; Vn induced formation of clusters where CK2 colocalized with Q03405 and alpha(v)beta(3) integrins . These results indicate that the uPA-dependent VSMC adhesion is a function of selective Vn phosphorylation by the ectoprotein kinase CK2 and suggest a regulatory role for Vn phosphorylation in the uPA-directed adhesive process . DB00013 plasminogen activator receptor ( CD87 ) expression of tumor-associated macrophages in ductal carcinoma in situ , breast cancer , and resident macrophages of normal breast tissue . Macrophages concentrate urokinase-type plasminogen activator ( uPA ) at the cell surface by expressing urokinase receptors ( Q03405 ) in order to focus the pericellular space plasminogen-dependent proteolysis important in matrix remodeling and cell movement . This study examines the Q03405 levels of tumor-associated macrophages ( TAM ) of invasive breast carcinomas , of TAMs from ductal carcinoma in situ ( DCIS ) and of macrophages derived from normal ( non-tumor ) breast tissue . TAMs from invasive breast carcinomas ( n = 30 ) , from DCIS ( n = 12 ) , and macrophages from normal breast tissue ( n = 30 ) were cultured and immunocytochemically phenotyped by using a panel of antibodies . DB00013 receptor levels were determined by Western blot analysis and in cell-free supernatants by enzyme-linked immunosorbent assay . DB00013 receptor cell surface fluorescence intensity was determined by FACS and by confocal laser scan microscopy . DB00013 -receptor mRNA was detected by in situ hybridization . TAMs of invasive breast carcinomas and of DCIS possess significantly elevated Q03405 levels compared with macrophages derived from normal breast tissue . CONCLUSIONS : activated macrophages with elevated Q03405 levels belong to inflammatory areas in close vicinity of infiltrating and non-infiltrating ( DCIS ) tumor cells . Blood monocytes that possess elevated Q03405 -levels may be selectively recruited from the bloodstream to inflammatory sites close to carcinoma cells , and/or breast cancer and precursor lesions may induce elevated Q03405 -levels in TAMs by paracrine interactions . DB00013 receptor up-regulation in head and neck squamous cell carcinoma . BACKGROUND : P00749 is important for matrix degradation and motility of cancer cells . For effective invasion , urokinase has to be associated with its cell surface receptor.(1) METHODS : We analyzed 33 head and neck squamous cell carcinomas ( hnSCC ) and 14 mucosal tissue samples for the expression of urokinase receptor using Northern hybridization and correlated expression levels to clinical and histopathologic data . DB00013 expression was determined by fibrin zymography . RESULTS : The expression of urokinase receptor is significantly increased in hnSCC compared with adjacent mucosa . Expression levels in primary tumors show no statistically significant correlations to T staging , metastasis , recurrence , or differentiation stage of the resected tumors . Furthermore , there was no correlation between urokinase and urokinase receptor expression levels in SCC samples . CONCLUSIONS : DB00013 receptor expression is increased in hnSCC , but it is not useful as a prognostic marker for the metastatic behavior of primary tumors . Comparison of our data with previously published reports is discussed . DB00013 -receptor-mediated phenotypic changes in vascular smooth muscle cells require the involvement of membrane rafts . The cholesterol-enriched membrane microdomains lipid rafts play a key role in cell activation by recruiting and excluding specific signalling components of cell-surface receptors upon receptor engagement . Our previous studies have demonstrated that the P06744 (glycosylphosphatidylinositol)-linked Q03405 [ uPA ( urokinase-type plasminogen activator ) receptor ] , which can be found in lipid rafts and in non-raft fractions , can mediate the differentiation of VSMCs ( vascular smooth muscle cells ) towards a pathophysiological de-differentiated phenotype . However , the mechanism by which Q03405 and its ligand uPA regulate VSMC phenotypic changes is not known . In the present study , we provide evidence that the molecular machinery of Q03405 -mediated VSMC differentiation employs lipid rafts . We show that the disruption of rafts in VSMCs by membrane cholesterol depletion using O95822 ( methyl-beta-cyclodextrin ) or filipin leads to the up-regulation of Q03405 and cell de-differentiation . Q03405 silencing by means of interfering RNA resulted in an increased expression of contractile proteins . Consequently , disruption of lipid rafts impaired the expression of these proteins and transcriptional activity of related genes . We provide evidence that this effect was mediated by Q03405 . Similar effects were observed in VSMCs isolated from Cav1Z(-/-) ( caveolin-1-deficient ) mice . Despite the level of Q03405 being significantly higher after the disruption of the rafts , uPA/ Q03405 -dependent cell migration was impaired . However , caveolin-1 deficiency impaired only Q03405 -dependent cell proliferation , whereas cell migration was strongly up-regulated in these cells . Our results provide evidence that rafts are required in the regulation of Q03405 -mediated VSMC phenotypic modulations . These findings suggest further that , in the context of uPA/ Q03405 -dependent processes , caveolae-associated and non-associated rafts represent different signalling membrane domains . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Adult patients with congenital adrenal hyperplasia have elevated blood pressure but otherwise a normal cardiovascular risk profile . OBJECTIVE : Treatment with glucocorticoids and mineralocorticoids has changed congenital adrenal hyperplasia ( CAH ) from a fatal to a chronic lifelong disease . Long-term treatment , in particular the chronic (over-)treatment with glucocorticoids , may have an adverse effect on the cardiovascular risk profile in adult CAH patients . The objective of this study was to evaluate the cardiovascular risk profile of adult CAH patients . DESIGN : Case-control study . PATIENTS AND MEASUREMENTS : In this case-control study the cardiovascular risk profile of 27 adult CAH patients and 27 controls , matched for age , sex and body mass index was evaluated by measuring ambulatory 24-hour blood pressure , insulin sensitivity ( HOMA-IR ) , lipid profiles , albuminuria and circulating cardiovascular risk markers ( P05121 , tPA , uPA , tPA/ P05121 complex , hsCRP , adiponectin , P05231 , Q14116 and leptin ) . RESULTS : 24-Hour systolic ( 126.3 mmHg±15.5 vs 124.8 mmHg±15.1 in controls , P = 0.019 ) and diastolic ( 76.4 mmHg±12.7 vs 73.5 mmHg±12.4 in controls , P < 0.001 ) blood pressure was significantly elevated in CAH patients compared to the control population . CAH patients had higher HDL cholesterol levels ( P < 0.01 ) , lower hsCRP levels ( P = 0.03 ) and there was a trend toward elevated adiponectin levels compared to controls . Other cardiovascular risk factors were similar in both groups . CONCLUSION : Adult CAH patients have higher ambulatory blood pressure compared to healthy matched controls . Other cardiovascular risk markers did not differ , while HDL-cholesterol , hsCRP and adiponectin levels tended to be more favorable . Tissue plasminogen activator and urokinase mediate the binding of DB00142 -plasminogen to plasma fibrin I . Evidence for new binding sites in plasmin-degraded fibrin I . The effect of tissue plasminogen activator ( TPA ) or urokinase on the specific binding of human DB00142 -plasminogen to fibrin I formed in plasma by clotting with Reptilase was studied using 125I-plasminogen and 131I-fibrinogen . In the absence of TPA , small amounts of plasminogen were bound to fibrin I . TPA induced binding of plasminogen to plasma fibrin I that was dependent upon the concentrations of TPA and plasminogen as well as upon the time of incubation . P00747 binding occurred in association with fibrin clot lysis and the formation in the clot supernatant of alpha 2-plasmin inhibitor-plasmin complexes . DB00013 also induced binding of plasminogen to plasma fibrin I that was concentration- and time-dependent . The molecular form of plasminogen bound to the fibrin I plasma clot was identified as DB00142 -plasminogen by dodecyl sulfate-polyacrylamide gel electrophoresis and by fast performance liquid chromatography . Further studies demonstrated that fibrin I formed from fibrinogen that had been progressively degraded by plasmin-bound DB00142 -plasminogen . The mole ratio of plasminogen bound increased with the time of plasmin digestion . DB00142 -plasminogen did not bind to fibrin I formed from fibrinogen progressively digested by human leukocyte elastase , thereby demonstrating the specificity of plasmin . These studies demonstrate that plasminogen activators regulate the binding of DB00142 -plasminogen to fibrin I by catalyzing plasmin-mediated modifications in the fibrin substrate . Anti-metastatic activities of Antrodia camphorata against human breast cancer cells mediated through suppression of the MAPK signaling pathway . The fermented culture broth of Antrodia camphorata ( A. camphorata ) has been shown to promote cell cycle arrest and apoptosis of human estrogen-nonresponsive MDA-MB-231 cells . Herein , we demonstrate that non-cytotoxic concentrations ( 20-80 μg/mL ) of A. camphorata markedly inhibited the invasion/migration of highly metastatic MDA-MB-231 cells as shown by an in vitro transwell and a wound-healing repair assay . The results of a gelatin zymography assay showed that A. camphorata suppressed the activity of matrix metalloproteinase ( MMP ) -9 and urokinase plasminogen activator ( uPA ) . Western blot results demonstrated that treatment with A. camphorata decreased the expression of P14780 , P08253 , uPA , uPA receptor ( Q03405 ) and vascular endothelial growth factor ( P15692 ) ; while the expression of the endogenous inhibitors of these proteins , i.e. , tissue inhibitors of MMP ( P01033 and P16035 ) , and plasminogen activator inhibitor ( P05121 ) -1 , increased . Further investigation revealed that A. camphorata suppressed the phosphorylation of P27361 /2 , p38 , and P45983 /2 . A. camphorata treatment also led to a dose-dependent inhibition on NF-κB binding and activation . This is the first report confirming the anti-metastatic activity of this potentially beneficial mushroom against human breast cancer . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . DB00013 induces survival or pro-apoptotic signals in human mesangial cells depending on the apoptotic stimulus . Deregulated apoptosis of MCs ( mesangial cells ) is associated with a number of kidney diseases including end-stage diabetic nephropathy . Cell death by apoptosis is a tightly orchestrated event , whose mechanisms are not completely defined . In the present study we show that the uPA ( urokinase-type plasminogen activator ) / Q03405 ( uPA receptor ) system can initiate both cell survival and pro-apoptotic signals in human MCs in response to different apoptotic stimuli . uPA abrogated MC apoptosis induced by serum withdrawal conditions and enhanced apoptosis initiated in MCs by high glucose . Effects of uPA were independent of its proteolytic activity and required Q03405 for both pro- and anti-apoptotic effects . Studies on the Q03405 interactome provide evidence that the opposing effects of uPA were directed via different Q03405 -interacting transmembrane partners . Exposure of MCs to RGD ( DB00125 - DB00145 - DB00128 ) peptide led to abrogation of the anti-apoptotic effect of uPA , which implies involvement of integrins in this process . A pro-apoptotic effect of uPA under high-glucose conditions was mediated via association of Q03405 and the cation-independent M6P ( mannose-6-phosphate ) / P11717 ( insulin-like growth factor 2 receptor ) . Both receptors were co-precipitated and co-localized in MCs . Studies on the underlying signalling indicate that the P27361 /2 ( extracellular-signal-regulated kinase 1/2 ) , Akt and Q92934 ( Bcl-2/Bcl-X(L)-antagonist , causing cell death ) protein were involved in regulation of apoptosis by uPA in MCs . P11717 mediated Q92934 perinuclear localization during apoptosis initiated by uPA and high glucose . In conclusion , we provide evidence that , in MCs , the uPA/ Q03405 system regulates survival/apoptosis processes in a stimulus-specific fashion via a mitochondria-dependent mechanism and that Q92934 protein serves as a downstream molecule . P00747 assay by means of the lysis time method . The lysis time method for the determination of plasminogen has been investigated using plasminogen-free thrombin and fibrinogen preparations . The experiments have shown that the lysis of a fibrin clot is the result of two consecutive reactions : the formation of fibrin which proceeds as a first order reaction and the degradation of fibrin which proceeds as a zero order reaction . P00747 is activated in a separate reaction . If the rate of the fibrin formation is much greater than the rate of degradation , the lysis of the fibrin clot is approximately of zero order in fibrin . The lysis time will then be inversely proportional to the plasmin concentration and proportional to the fibrinogen concentration . In a double logaritmic system the correlation between lysis time and plasmin activity is a straight line with a slope of 135 degrees . P00747 is rapidly activated with streptokinase . Maximal activation is obtained only with a certain streptokinase concentration . Higher concentrations inactivate plasmin and with lower concentrations , the maximal activity is never reached . A spontaneous inactivation is seen after about 30 minutes . With urokinase , a higher maximal plasminogen activity is obtained than with streptokinase . DB00013 in higher concentrations does not inactivate plasmin . A standard assay for determination of plasminogen by the lysis time method has been worked out and is based on these results . DB00013 plasminogen activator and its inhibitor , P05121 , as prognostic markers in breast cancer : from pilot to level 1 evidence studies . BACKGROUND : For optimum management of patients with cancer , accurate assessment of prognosis is essential . The primary determinant of outcome in malignancy is the formation of distant metastases . DB00013 plasminogen activator ( uPA ) is a serine protease causally involved in invasion and metastasis . CONTENT : Data from model systems show that uPA is unequivocally involved in cancer dissemination . Consistent with its role in metastasis , multiple independent groups have shown that high uPA concentrations in primary breast cancers correlate with poor prognosis . For determining outcome , the prognostic impact of uPA was both independent of traditionally used factors and prognostic in patients with axillary node-negative disease . Paradoxically , high concentrations of plasminogen activator inhibitor ( P05121 ) , an endogenous inhibitor of uPA , also correlate with poor prognosis in patients with breast cancer , including the subgroup with node-negative disease . The prognostic value of uPA/ P05121 in axillary node-negative breast cancer patients was recently confirmed in both a prospective randomized trial and a pooled analysis , i.e. , two different level 1 evidence ( LOE-1 ) studies . CONCLUSIONS : uPA and P05121 are among the first biological prognostic factors to have their clinical value validated using LOE-1 evidence studies . Determination of these analytes may help identify low-risk node-negative breast cancer patients for whom adjuvant chemotherapy is unnecessary . Association of polymorphisms in the genes of the urokinase plasminogen activation system with susceptibility to and severity of non-small cell lung cancer . BACKGROUND : DB00013 plasminogen activating ( uPA ) system is implicated in neoplastic progression . High tissue levels of uPA system components correlate with a poor prognosis in lung cancer . The present study examined the single nucleotide polymorphisms ( SNPs ) of uPA and the corresponding receptor , Q03405 , for exploring their roles in non-small cell lung cancer ( NSCLC ) . METHODS : The allele frequencies and genotype distributions of uPA rs4065 C/T and Q03405 rs344781 ( -516 T/C ) among 375 NSCLC cases and 380 healthy controls were examined using polymerase chain reaction-restriction fragment-length polymorphism ( PCR-RFLP ) analysis . Putative association between the above SNPs and clinicopathological characteristics of NSCLC were also analyzed . RESULTS : The genotype frequencies of the variant homozygotes of uPA and Q03405 were significantly different between NSCLC and control subjects . Significant association was also observed between the examined genotypes and disease stage of NSCLC . Logistic regression analysis revealed that individuals with uPA rs4065 TT genotype have higher odds ratios ( ORs ) for lung cancer . Whereas , subjects with Q03405 -344781 CC genotype have lower ORs for lung cancer . The patients carrying a homozygous TT genotype at uPA rs4065 , or at least a T allele at Q03405 -344781 ( -516 ) , had a tendency to develop advanced disease . CONCLUSIONS : Our results revealed that genetic polymorphisms of the uPA rs4065 C/T and Q03405 rs344781 ( -516 T/C ) were associated with the susceptibility and severity of NSCLC . An anti-urokinase plasminogen activator receptor antibody ( ATN-658 ) blocks prostate cancer invasion , migration , growth , and experimental skeletal metastasis in vitro and in vivo . DB00013 plasminogen activator receptor ( Q03405 ) is a multidomain protein that plays important roles in the growth , invasion , and metastasis of a number of cancers . In the present study , we examined the effects of administration of a monoclonal anti- Q03405 antibody ( ATN-658 ) on prostate cancer progression in vitro and in vivo . We examined the effect of treatment of ATN-658 on human prostate cancer cell invasion , migration , proliferation , and regulation of intracellular signaling pathways . For in vivo studies , PC-3 cells ( 1 x 10(6) ) were inoculated into the right flank of male Balb C nu/nu mice through subcutaneous or through intratibial route ( 2 x 10(5) ) of male Fox Chase severe combined immunodeficient mice to monitor the effect on tumor growth and skeletal metastasis . Treatment with ATN-658 resulted in a significant dose-dependent decrease in PC-3 cell invasion and migration without affecting cell doubling time . Western blot analysis showed that ATN-658 treatment decreased the phosphorylation of serine/threonine protein kinase B ( AKT ) , mitogen-activated protein kinase ( MAPK ) , and focal adhesion kinase ( Q05397 ) without affecting AKT , MAPK , and Q05397 total protein expression . In in vivo studies , ATN-658 caused a significant decrease in tumor volume and a marked reduction in skeletal lesions as determined by Faxitron x-ray and micro-computed tomography . Immunohistochemical analysis of subcutaneous and tibial tumors showed a marked decrease in the levels of expression of pAKT , pMAPK , and pFAK , consistent with the in vitro observations . Results from these studies provide compelling evidence for the continued development of ATN-658 as a potential therapeutic agent for the treatment of prostate and other cancers expressing Q03405 . DB00013 -induced activation of the P40189 /Tyk2/Stat3 pathway mediates a pro-inflammatory effect in human mesangial cells via expression of the anaphylatoxin C5a receptor . Glomerular mesangial cells ( MCs ) are central to the pathogenesis of progressive glomeruli-associated renal diseases . However , molecular mechanisms underlying changes in MC functions still remain poorly understood . Here , we show that in MCs , the urokinase-type plasminogen activator ( uPA ) induces , via its specific receptor ( Q03405 , CD87 ) , upregulated expression of the complement anaphylatoxin C5a receptor ( P21730 , CD88 ) , and modulates C5a-dependent functional responses . This effect is mediated via the interaction of the uPA-specific receptor ( Q03405 , CD87 ) and P40189 , a signal transducing subunit of the receptor complexes for the P05231 cytokine family . The Janus kinase Tyk2 and the transcription factor Stat3 serve as downstream components in the signaling cascade resulting in upregulation of P21730 expression . In vivo , expression of P21730 and Q03405 was increased in the mesangium of wild-type mice in a lipopolysaccharide ( LPS ) -induced model of inflammation , whereas in Q03405 (-/-) animals P21730 expression remained unchanged . This is the first demonstration in vitro and in vivo that uPA acts in MCs as a modulator of immune responses via control of immune-competent receptors . The data suggest a novel role for uPA/ Q03405 in glomeruli-associated renal failure via a signaling cross-talk between the fibrinolytic and immune systems . DB00013 binding and catabolism by Hep G2 cells is plasminogen activator inhibitor-1 dependent , analogous to interactions of tissue-type plasminogen activator with these cells . The adherent human hepatoma cell line Hep G2 exhibits receptor mediated endocytosis and catabolism of tissue-type plasminogen activator.plasminogen activator inhibitor type-1 ( t-PA. P05121 ) complexes formed when exogenous t-PA combines with endogenous P05121 in the extracellular matrix . To determine whether the other major PA , urokinase ( u-PA ) , which also complexes with P05121 , is metabolised via the same mechanism , 125I-labelled high ( hmw ) and low ( lmw ) molecular weight forms of u-PA were incubated with Hep G2 cells at 4 degrees C for 2 hr in the absence and presence of a 100-fold excess of unlabelled ligand in order to detect specific binding . Both hmw and lmw 125I-u-PA formed complexes with P05121 and these bound specifically and with high affinity ( apparent Kd 3.9 and 4.1 nM , with Bmax 78 x 10(3) and 83 x 10(3) binding sites/cell respectively ) . Binding by each form of radiolabelled u-PA was inhibited in a dose-dependent fashion by unlabelled t-PA , hmw-u-PA , lmw-u-PA , and by monoclonal anti- P05121 antibody . At 37 degrees C , bound hmw and lmw 125I-u-PA. P05121 complexes were internalised and degraded rapidly . These findings indicate that the specificity of the previously described receptor which mediates P05121 dependent catabolism of t-PA by Hep G2 cells extends to complexes of u-PA with this inhibitor . Q03405 and cathepsin B inhibition enhanced radiation-induced apoptosis in gliomainitiating cells . Glioblastomas present as diffuse tumors with invasion into normal brain tissue and frequently recur or progress after radiation as focal masses because of glioma-initiating cells . The role of the urokinase-type plasminogen activator receptor ( Q03405 ) and cathepsin B in stem-like phenotype has been extensively studied in several solid tumors . In the present study , we demonstrated that selection of glioma-initiating cells using CD133 expression leads to a specific enrichment of CD133(+) cells in both U87 and 4910 cells . In addition , CD133(+) cells exhibited a considerable amount of other stem cell markers , such as P48681 and Sox-2 . Radiation treatment significantly enhanced Q03405 and cathepsin B levels in glioma-initiating cells . To downregulate radiation-induced Q03405 and cathepsin B expression , we used a bicistronic shRNA construct that simultaneously targets both Q03405 and cathepsin B ( pCU ) . Downregulation of Q03405 and cathepsin B using pCU decreased radiation-enhanced Q03405 and cathepsin B levels and caused DNA damage-induced apoptosis in glioma cell lines and glioma-initiating cells . The most striking finding of this study is that knockdown of Q03405 and cathepsin B inhibited ongoing transcription by suppressing BrUTP incorporation at γ P16104 foci . In addition , Q03405 and cathepsin B gene silencing inversely regulated survivin and P16104 expression in both glioma cells and glioma-initiating cells . Pretreatment with pCU reduced radiation-enhanced expression of Q03405 , cathepsin B , and survivin and enhanced DNA damage in pre-established glioma in nude mice . Taken together , our in vitro and in vivo findings suggest that Q03405 and cathepsin B inhibition might serve as an adjunct to radiation therapy to target glioma-initiating cells and , therefore , for the treatment of glioma . G protein-coupled receptor kinase-3-deficient mice exhibit WHIM syndrome features and attenuated inflammatory responses . Chemokine receptor interactions coordinate leukocyte migration in inflammation . Chemokine receptors are GPCRs that when activated , are phosphorylated by GRKs to turn off G protein-mediated signaling yet recruit additional signaling machinery . Recently , P35626 was identified as a negative regulator of P48061 / P61073 signaling that is defective in human WHIM syndrome . Here , we report that P35626 -/- mice exhibit numerous features of human WHIM , such as impaired P48061 -mediated desensitization , enhanced P61073 signaling to P29323 activation , altered granulocyte migration , and a mild myelokathexis . Moreover , P35626 -/- protects mice from two acute models of inflammatory arthritis ( K/BxN serum transfer and CAIA ) . In these granulocyte-dependent disease models , protection of P35626 -/- mice is mediated by retention of cells in the marrow , fewer circulating granulocytes in the peripheral blood , and reduced granulocytes in the joints during active inflammation . In contrast to WHIM , P35626 -/- mice have minimal hypogammaglobulinemia and a peripheral leukocytosis with increased lymphocytes and absent neutropenia . Thus , we conclude that the loss of P35626 -mediated regulation of P48061 / P61073 signaling contributes to some , but not all , of the complete WHIM phenotype and that P35626 inhibition may be beneficial in the treatment of inflammatory arthritis . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . P00747 potentiates thrombin cytotoxicity and contributes to pathology of intracerebral hemorrhage in rats . Thrombin and plasmin are serine proteases involved in blood coagulation and fibrinolysis , whose precursors are circulating in blood stream . These blood-derived proteases might play important roles in the pathogenesis of intracerebral hemorrhage by acting on brain parenchymal cells . We previously reported that thrombin induced delayed neuronal injury through extracellular signal-regulated kinase ( P29323 ) -dependent pathways . Here , we investigated potential cytotoxic actions of plasminogen , a precursor protein of plasmin , using slice cultures prepared from neonatal rat brain and intracortical microinjection model in adult rats . Although plasminogen alone did not evoke prominent neuronal injury , plasminogen caused significant neuronal injury when combined with a moderate concentration of thrombin ( 30 U/mL ) in the cerebral cortex of slice cultures . The cortical injury was prevented by tranexamic acid and aprotinin . The combined neurotoxicity of thrombin and plasminogen was also prevented by PD98059 , an inhibitor of P29323 pathway , as well as by other agents that have been shown to prevent cortical injury induced by a higher concentration ( 100 U/mL ) of thrombin alone . Extracellular signal-regulated kinase phosphorylation after plasminogen exposure was localized in cortical astrocytes . Moreover , microinjection of plasminogen in vivo potentiated thrombin-induced cortical injury , and inhibition of plasmin ameliorated hemorrhage-induced neuronal loss in the cerebral cortex . These results suggest that plasminogen/plasmin system augmenting thrombin neurotoxicity participates in hemorrhagic cortical injury . Epithelial and stromal cell urokinase plasminogen activator receptor expression differentially correlates with survival in rectal cancer stages B and C patients . DB00013 plasminogen activator receptor ( Q03405 ) has been proposed as a potential prognostic factor for colorectal cancer ( CRC ) patient survival . However , CRC Q03405 expression remains controversial , especially regarding cell types where Q03405 is overexpressed ( e.g. , epithelium ( uPARE ) or stroma-associated cells ( uPARS ) ) and associated prognostic relevance . In this study , two epitope-specific anti- Q03405 monoclonal antibodies ( MAbs ) could discriminate expression of uPARE from uPARS and were used to examine this association with survival of stages B and C rectal cancer ( RC ) patients . Using immunohistochemistry , MAbs # 3937 and R4 were used to discriminate uPARE from uPARS respectively in the central and invasive frontal regions of 170 stage B and 179 stage C RC specimens . Kaplan-Meier and Cox regression analyses were used to determine association with survival . Q03405 expression occurred in both epithelial and stromal compartments with differential expression observed in many cases , indicating uPARE and uPARS have different cellular roles . In the central and invasive frontal regions , uPARE was adversely associated with overall stage B survival ( HR = 1.9 ; p = 0.014 and HR = 1.5 ; p = 0.031 , respectively ) reproducing results from previous studies . uPARS at the invasive front was associated with longer stage C survival ( HR = 0.6 ; p = 0.007 ) , reflecting studies demonstrating that macrophage peritumoural accumulation is associated with longer survival . This study demonstrates that different Q03405 epitopes should be considered as being expressed on different cell types during tumour progression and at different stages in RC . Understanding how uPARE and uPARS expression affects survival is anticipated to be a useful clinical prognostic marker of stages B and C RC . Fibrinolytic effects of urokinase and heparin in acute pulmonary embolism : a randomized clinical trial . Dissolution of pulmonary emboli with heparin and urokinase is ascribed , respectively , to anticoagulation and fibrinolysis . Since truly independent assessment of these effects in man is lacking , we administered each drug alone . DB09222 and plasminogen plasma levels and the resolution of pulmonary emboli were measured in three randomized groups of 10 patients each : groups A and C infused with small repeated doses of urokinase and a large single dose of urokinase , respectively , and group B who received heparin . After 6 h of treatment , fibrinogen fell in all the groups , while , after 12 h , remained equally reduced in groups A and B and declined further in group C . P00747 behaved similarly . Up to 60 h , statistical analysis showed that these effects were related to timing and amounts of urokinase and heparin infusion . These observations suggest that heparin may induce a lytic state . As to signs of pulmonary emboli resolution , no differences between groups were found in lung perfusion and gas exchange recovery at any time ( from 1 day to 1 year ) and in pulmonary artery pressure reduction at 1 week . The greater angiographic and scintigraphic recovery observed with urokinase , versus heparin alone , after 1 day of treatment in the DB00013 Pulmonary Embolism Trial may be ascribed to a synergistic effect with urokinase of heparin administered during the diagnostic work-out . The indications of heparin and urokinase should be evaluated in the light of these results . DB11320 modulates multiple functional activities of monocyte-derived dendritic cell subsets via histamine receptor 2 . Expression of CD1a proteins in human monocyte-derived dendritic cells ( DCs ) specifies functionally distinct subsets with different inflammatory properties . DB11320 is recognized as an inflammatory mediator released by various cell types including DCs . The diverse biological effects of histamine are mediated by G-protein-coupled histamine receptors ( HRs ) , which are able to modulate the functional activities of DC subsets . The goal of the present study was to compare the expression and activity of HRs in the CD1a(-) and CD1a(+) monocyte-derived DC subsets and to test the effects of histamine on the differentiation , activation and functional activities of these subsets . We show that P25021 is present at high levels in both DC subsets , whereas P35367 and Q9H3N8 are expressed in a subset-specific manner . DB11320 shifts DC differentiation to the development of CD1a(-) DCs and modulates DC activation through its inhibitory effect on CD1a(+) DC differentiation . DB11320 -induced reduction of CD1a(+) DCs is associated with increased secretion of P05231 and P22301 , up-regulation of a typical combination of chemokines , expression C5aR1 by the CD1a(-) DC subset and enhanced migration of both activated DC subsets supported by the production of P14780 and P39900 enzymes . All these effects were shown to be mediated in a P25021 -specific manner as revealed by the specific antagonist of the receptor . As P25021 is expressed at high levels in both DC subsets , we propose that it may dominate the regulation of multiple DC functions . In contrast , P35367 and Q9H3N8 with opposing subset-related expression may have a regulatory or fine-tuning role in histamine-induced functional activities . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Treatment of thoracic multiloculated empyemas with intracavitary urokinase : a prospective study . The authors prospectively treated 10 consecutive patients with multiloculated empyemas with intracavitary instillation of urokinase via a percutaneous drainage catheter . DB00013 ( 100,000 IU ) in 100 mL of 5 % dextrose in water was instilled into the pleural cavity via a percutaneous drainage catheter . After overnight clamping , the catheter was opened and the empyema drained with use of negative suction ( 20 cm H2O ) . Intermittent irrigation of the catheter with normal saline was performed to prevent clogging of the catheter . Complete drainage of multiloculated empyemas was accomplished in nine patients by means of intracavitary instillation of urokinase via a single 8-F catheter . One patient showed complete drainage of multiloculated empyema , but recurrent empyema appeared in the site of a previous tube thoracostomy . A total of 100,000-700,000 IU ( mean , 400,000 IU ) of urokinase were needed for complete drainage in all patients . P00747 and fibrin degradation product levels in empyema fluid were determined before instillation of urokinase to demonstrate any fibrinolytic action . No complications occurred .

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interacts_with DB08895?

multiple_choice

DB08895 in kidney transplantation . INTRODUCTION : This review will discuss the mechanism of action and important kidney transplant clinical trial data for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . AREAS COVERED : Successful kidney transplantation requires adequate immunosuppression . Current maintenance immunosuppressive protocols which rely on calcineurin inhibitors have long-term nephrotoxicity and negative impact on cardiometabolic risk factors . JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared to control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . EXPERT OPINION : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . Hyperecho-turbo spin-echo sequences at 3T : clinical application in neuroradiology . BACKGROUND AND PURPOSE : Hyperecho-turbo spin-echo ( hyperTSE ) sequences were developed to reduce the specific absorption rate ( SAR ) , especially at high fields such as 3T and above . The purpose of this study was to quantitatively and qualitatively assess the detection of neuroradiologic pathologies by hyperTSE in comparison with standard turbo spin-echo ( TSE180 degrees ) sequences . MATERIALS AND METHODS : TSE180 degrees and hyperTSE images with parameters adapted for equal P24752 contrast were acquired on a 3T whole-body system in 51 patients with 54 cerebral pathologies . Region-of-interest analysis was performed of signal intensities of pathologies , normal white and gray matter , P04141 , and the SD of noise . Signal intensity-to-noise ratios ( SNRs ) and contrast-to-noise ratios ( CNRs ) for healthy tissues and pathologies were determined . A qualitative rating concerning artifacts , lesion conspicuity , and image quality was performed by 2 experienced neuroradiologists . RESULTS : HyperTSE sequences were equivalent to standard TSE180 degrees sequences for the P21554 of pathologies and of the contrast between gray and white matter . The SNR of gray and white matter and P04141 were also the same . The CNRs of the pathologies in hyperTSE and TSE180 degrees images were strongly correlated with each other ( r = 0.93 , P = .001 ) . The visual rating of images revealed no significant differences between hyperTSE and TSE180 degrees . CONCLUSION : HyperTSE sequences proved to be qualitatively and quantitatively equivalent to TSE180 degrees sequences in the detection of high- and low-signal-intensity lesions . They provide equal P21554 of pathologies and of gray minus white matter and reduce the imaging restrictions of conventional TSE180 degrees imposed by SAR limitations at 3T . The p65 ( RelA ) subunit of NF-kappaB interacts with the histone deacetylase ( HDAC ) corepressors Q13547 and Q92769 to negatively regulate gene expression . Regulation of NF-kappaB transactivation function is controlled at several levels , including interactions with coactivator proteins . Here we show that the transactivation function of NF-kappaB is also regulated through interaction of the p65 ( RelA ) subunit with histone deacetylase ( HDAC ) corepressor proteins . Our results show that inhibition of HDAC activity with trichostatin A ( P32119 ) results in an increase in both basal and induced expression of an integrated NF-kappaB-dependent reporter gene . Chromatin immunoprecipitation ( ChIP ) assays show that P32119 treatment causes hyperacetylation of the wild-type integrated NF-kappaB-dependent reporter but not of a mutant version in which the NF-kappaB binding sites were mutated . Expression of Q13547 and Q92769 repressed tumor necrosis factor ( P01375 ) -induced NF-kappaB-dependent gene expression . Consistent with this , we show that Q13547 and Q92769 target NF-kappaB through a direct association of Q13547 with the Rel homology domain of p65 . Q92769 does not interact with NF-kappaB directly but can regulate NF-kappaB activity through its association with Q13547 . Finally , we show that inhibition of HDAC activity with P32119 causes an increase in both basal and P01375 -induced expression of the NF-kappaB-regulated interleukin-8 ( P10145 ) gene . Similar to the wild-type integrated NF-kappaB-dependent reporter , ChIP assays showed that P32119 treatment resulted in hyperacetylation of the P10145 promoter . These data indicate that the transactivation function of NF-kappaB is regulated in part through its association with HDAC corepressor proteins . Moreover , it suggests that the association of NF-kappaB with the Q13547 and Q92769 corepressor proteins functions to repress expression of NF-kappaB-regulated genes as well as to control the induced level of expression of these genes . Involvement of P50406 receptors in nigro-striatal function in rodents . 4-Amino-N- ( 2,4 bis-methylamino-pyrimidin-4-yl ) benzene sulphonamide ( Ro 04-6790 ) is a potent , selective and competitive antagonist for the P50406 receptor which can be detected in the cerebro-spinal fluid ( P04141 ) of rats following intraperitoneal administration . Since P50406 receptor mRNA and P50406 receptor-like immunoreactivity have been shown to be present in the striatum , the purpose of the present study was to evaluate the effect of P50406 receptor antagonism on haloperidol- and P35240 23390-induced catalepsy in mice and on the turning behaviour of rats with unilateral 6-hydroxydopamine ( 6-OHDA ) lesions of the medial forebrain bundle . Ro 04-6790 ( 3 , 10 and 30 mg kg(-1) i.p. ) did not induce catalepsy and had no effect on catalepsy induced by either haloperidol or P35240 23390 . Ro 04-6790 ( 3 , 10 and 30 mg kg(-1) i.p. ) did not itself induce rotational behaviour in rats with unilateral 6-hydroxydopamine ( 6-OHDA ) lesions of the medial forebrain bundle nor did it affect the rotational behaviour induced by either L-Dopa or amphetamine . P50406 receptor antagonism inhibited the rotational behaviour of 6-OHDA lesioned rats induced by treatment with the muscarinic antagonists scopolamine and atropine . The data support earlier conclusions from experiments with antisense oligonucleotides that the P50406 receptor is involved in the control of acetylcholine neurotransmission in the rat brain . Kaempferol inhibits P05112 -induced P42226 activation by specifically targeting P52333 . P05112 is involved in several human diseases including allergies , autoimmunity , and cancer . Its effects are mainly mediated through the transcription factor P42226 . Therefore , investigation of compounds that regulate P42226 activation is of great interest for these diseases . Natural polyphenols are compounds reported to have therapeutic properties in diseases involving P05112 and P42226 . The aim of this study was to investigate the effect of these compounds in the activation of this transcription factor . We found that in hemopoietic cells from human and mouse origin , some flavonoids were able to inhibit the activation of P42226 by P05112 . To identify molecular mechanisms , we focused on kaempferol , the compound that showed the greatest inhibitory effect with the lowest cell toxicity . Treatment of cells with kaempferol did not affect activation of Src kinase by P05112 but did prevent the phosphorylation of P23458 and P52333 . Further enzymatic analysis demonstrated that kaempferol blocked the in vitro phosphorylation activity of P52333 without affecting P23458 , suggesting that it specifically targeted P52333 activity . Accordingly , kaempferol had no effect on P42226 activation in nonhemopoietic cell lines lacking P52333 , supporting its selective inhibition of P05112 responses through type I receptors expressing P52333 but not type II lacking this kinase . The inhibitory effect of kaempferol was also observed in P60568 but not P08700 -mediated responses and correlated with the inhibition of MLC proliferation . These findings reveal the potential use of kaempferol as a tool for selectively controlling cell responses to P05112 and , in general , P52333 -dependent responses . Astroglial P21554 cannabinoid receptors regulate leptin signaling in mouse brain astrocytes . Type-1 cannabinoid ( P21554 ) and leptin ( ObR ) receptors regulate metabolic and astroglial functions , but the potential links between the two systems in astrocytes were not investigated so far . Genetic and pharmacological manipulations of P21554 receptor expression and activity in cultured cortical and hypothalamic astrocytes demonstrated that cannabinoid signaling controls the levels of ObR expression . Lack of P21554 receptors also markedly impaired leptin-mediated activation of signal transducers and activators of transcription 3 and 5 ( P40763 and P42229 ) in astrocytes . In particular , P21554 deletion determined a basal overactivation of P42229 , thereby leading to the downregulation of ObR expression , and leptin failed to regulate P42229 -dependent glycogen storage in the absence of P21554 receptors . These results show that P21554 receptors directly interfere with leptin signaling and its ability to regulate glycogen storage , thereby representing a novel mechanism linking endocannabinoid and leptin signaling in the regulation of brain energy storage and neuronal functions . Structure-based design of novel anticancer agents . Recently identified agents that interact with cytoskeletal elements such as tubulin include synthetic spiroketal pyrans ( SPIKET ) and monotetrahydrofuran compounds ( COBRA compounds ) . SPIKET compounds target the spongistatin binding site of beta-tubulin and COBRA compounds target a unique binding cavity on alpha-tubulin . At nanomolar concentrations , the SPIKET compound SPIKET-P causes tubulin depolymerization and exhibits potent cytotoxic activity against cancer cells . COBRA-1 inhibits GTP-induced tubulin polymerization . Treatment of human breast cancer and brain tumor cells with COBRA-1 caused destruction of microtubule organization and apoptosis . Other studies have identified some promising protein tyrosine kinase inhibitors as anti-cancer agents . These include P00533 inhibitors such as the quinazoline derivative WHI-P97 and the leflunomide metabolite analog LFM-A12 . Both LFM-A12 and WHI-P97 inhibit the in vitro invasiveness of P00533 positive human breast cancer cells at micromolar concentrations and induce apoptotic cell death . Dimethoxyquinazoline compounds WHI-P131 and WHI-P154 inhibit tyrosine kinase P52333 in leukemia cells . Of particular interest is WHI-P131 , which inhibits P52333 but not P23458 , O60674 , P43405 , Q06187 , P07948 , or IRK at concentrations as high as 350 microM . Studies of Q06187 inhibitors showed that the leflunomide metabolite analog LFM-A13 inhibited Q06187 in leukemia and lymphoma cells . Consistent with the anti-apoptotic function of Q06187 , treatment of leukemic cells with LFM-A13 enhanced their sensitivity to chemotherapy-induced apoptosis . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Inhibitors of P11274 signalling interrupt the survival signal mediated by the micro-environment in mantle cell lymphoma . Several studies provide evidences for mantle cell lymphoma ( Q8WXI8 ) cell survival relying on B-cell receptor ( P11274 ) -mediated signalling pathways , whereas the nature of this activation is unknown . Significant progress in Q8WXI8 treatment is achieved through therapies targeting P11274 -associated kinases , i.e. , DB09053 and Fostamatinib , inhibitors of Q06187 and P43405 , respectively . Our study addresses survival signals emanating from the P11274 or the tumour environment and how inhibiting P11274 signalling effectors might impact these survival signals . We found that Q06187 was constitutively activated and that P43405 phosphorylation was highly increased and sustained upon P11274 activation of primary Q8WXI8 cells . Moreover , Q8WXI8 cells from leukaemic patients secreted high amount of IL-1β , P05231 , P10145 and P13501 . Activation of the P11274 induced ( i ) cell survival , ( ii ) P40763 activation and ( iii ) increased autocrine secretion of IL-1β , P05231 , P10145 , P13501 , P22301 , TNFα and P15692 . Specific inhibition of Q06187 by DB09053 or P43405 by Fostamatinib ( R406 ) reversed these protective effects and decreased both basal and P11274 -induced autocrine cytokine secretions associated with P40763 phosphorylation . Interestingly , targeting Q06187 and P43405 prevented and inhibited P11274 -induced Q8WXI8 cell adhesion to human bone marrow stromal cells ( HMSCs ) in short- and long-term co-culture . We demonstrated that P11274 -induced survival relies on autocrine secretion of IL-1β , TNFα and P13501 that might facilitate adhesion of Q8WXI8 cells to HMSC . Treatment with DB09053 or Fostamatinib blocked the chemotactic signal thus increasing apoptosis . Janus kinase inhibition with tofacitinib : changing the face of inflammatory bowel disease treatment . The advent of anti-Tumor Necrosis Factor ( P01375 ) therapy has changed the way of treating inflammatory bowel disease ( Q9UKU7 ) . However , primary and secondary failure are relatively frequent with all anti- P01375 agents , which are available only as parenteral agents . DB08895 is an oral janus kinase ( JAK ) inhibitor that inhibits JAK family kinase members , in particular P23458 and P52333 , achieving a broad limitation of inflammation by interfering with several cytokine receptors . It first proved its efficacy as an immunosuppressive regimen after renal transplantation , and was recently approved by the FDA for rheumatoid arthritis . First data in Q9UKU7 are promising , especially in ulcerative colitis . Ongoing clinical trials in both UC and Crohn 's disease ( CD ) are needed to further explore its efficacy in CD and to better assess its safety profile . Mechanism of inflammation in murine eosinophilic myocarditis produced by adoptive transfer with ovalbumin challenge . BACKGROUND : Interleukin ( IL ) -5 , RANTES and CC chemokine receptor 3 ( P51677 ) are essential for induction of eosinophil recruitment in organs , but the precise pathogenesis of eosinophilic myocarditis is still unclear . We investigated the relationships between these cytokines and receptors in the development of inflammation in murine myocarditis produced by adoptive transfer , with reference to eosinophil infiltration and signal transduction . METHODS : The splenocytes from male donor DBA/2 mice were separated after ovalbumin ( OVA ) sensitization . These cells had a P01730 /CD8 ratio of approximately 3.0 . Cells ( 2.0 x 10(7) ) were individually transfused to recipient adoptive male DBA/2 mice , and OVA challenge was performed serially . The heart and spleen of the recipient were analyzed to determine the kinetics of P05113 , RANTES , P51677 and eosinophil production with simultaneous determination of P52333 ( P52333 ) mRNA . RESULTS : Approximately 85 % of recipient mice developed myocarditis ; 35 % had recognizable cell infiltration in the left ventricular endocardium , an effect which was absent in control mice . Eosinophilic myocarditis was usually associated with animals having several degenerative changes in myocardial cells , and P05113 , RANTES and P51677 expressions were usually present in these eosinophils ( p < 0.05 ) . P51677 and P52333 mRNAs were detected in the spleens and hearts of recipient animals providing histological evidence for kinetics related to eosinophil infiltration . CONCLUSION : The murine model of adoptive transferred myocarditis is suitable for studying the mechanism of eosinophilic myocarditis . A unique pathogenesis of this disorder may be controlled by the synergism of P01730 dominancy in the donor and JAK- P35610 signaling in the recipient , which may cause recruitment of eosinophils into heart lesions . DB08895 . DB08895 ( CP-690,550 ; CP-690550 ; CP690550 ) , an orally active immunosuppressant , is being developed by Pfizer for the treatment of rheumatoid arthritis , inflammatory bowel disease , dry eyes , ankylosing spondylitis , psoriasis , psoriatic arthritis , and for the prevention of transplant rejection . DB08895 specifically inhibits Janus activated kinase 3 ( P52333 ) , which has a pivotal role in cytokine signal transduction that governs lymphocyte survival , proliferation , differentiation , and apoptosis . This review discusses the key development milestones and therapeutic trials of this drug . Synthesis and biological evaluation of novel pyrrolidine-2,5-dione derivatives as potential antidepressant agents . Part 1 . A series of 3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by (1)H NMR , (13)C NMR and P19957 -HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5- Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists . P04035 inhibitors deplete circulating classical and non-classical monocytes following human heart transplantation . BACKGROUND : Monocytes mediate immune responses following solid organ transplantation via cytokine secretion and differentiation to macrophage/dendritic cell lineages . To date , the pleiotropic immunomodulatory effect of statins on human monocytes following human heart transplantation has yet to be elucidated . This study was designed to assess the effects of statin administration on the monocyte repertoire . METHODS : 108 patients were recruited into the study . Clinical data were collected from patients ' notes . Peripheral blood immunophenotype was determined via flow cytometry ( using CD11c , P08571 , CD16 , CD49d , CD64 , P33681 and CD195 ) . RESULTS : There were fewer circulating classical ( p=0.0001 ) and non-classical ( p=0.0013 ) monocytes in patients treated with a statin . CD64 expression was down-regulated ( p=0.011 and p=0.049 ) whereas CD49d expression was up-regulated ( p=0.004 and p=0.022 ) on classical and non-classical monocytes in this group . Patients receiving DB01076 had fewer circulating classical monocytes ( p=0.001 ) compared to patients administered DB00175 . Patients receiving DB00175 had fewer circulating non-classical monocytes ( p=0.029 ) compared to patients administered DB01076 . DISCUSSION : Statin administration alters the circulating monocyte repertoire following heart transplantation , including population size , FcgammaRI and VLA-4 adhesion molecule expression . Furthermore , different statin treatments are associated with a selective depletion of macrophage or DC (re)generating monocytes . JAK inhibitors : treatment efficacy and safety profile in patients with psoriasis . Janus kinase ( JAK ) pathways are key mediators in the immunopathogenesis of psoriasis . Psoriasis treatment has evolved with the advent of targeted therapies , which inhibit specific components of the psoriasis proinflammatory cascade . JAK inhibitors have been studied in early phase trials for psoriasis patients , and the data are promising for these agents as potential treatment options . DB08895 , an oral or topically administered P23458 and P52333 inhibitor , and ruxolitinib , a topical P23458 and O60674 inhibitor , have been most extensively studied in psoriasis , and both improved clinical symptoms of psoriasis . Additional P23458 or P52333 inhibitors are being studied in clinical trials . In phase III trials for rheumatoid arthritis , tofacitinib was efficacious in patients with inadequate responses to tumor necrosis factor inhibitors , methotrexate monotherapy , or disease-modifying antirheumatic drugs . The results of phase III trials are pending for these therapies in psoriasis , and these agents may represent important alternatives for patients with inadequate responses to currently available agents . Further investigations with long-term clinical trials are necessary to verify their utility in psoriasis treatment and assess their safety in this patient population . Acute effects of sarpogrelate , a 5- Q13049 receptor antagonist on cytokine production in endotoxin shock model of rats . Serotonin ( 5-HT ) (2A) receptors are involved in cytokine production in infection or sepsis . Therefore , 5-HT(2A) receptor antagonist might be useful to treat sepsis . The present study investigates the effects of a 5-HT(2A) receptor antagonist , sarpogrelate on endotoxin shock . Catheters were inserted into the femoral artery and vein of Sprague-Dawley rats . First , sarpogrelate 0 ( control ) , 3 , or 10 mg/kg dissolved in 0.5 ml of distilled water has been given , followed by endotoxin 10 mg/kg in saline 0.5 ml 5 min later . Blood pressure , pulse rate and survival rate were monitored in 20 rats per dose . Blood gas and plasma cytokine concentrations were measured in 8 rats per dose . In four rats each of sarpogrelate 0 , 3 , or 10 mg/kg , and sham operation , the lung histology was examined . Zero , 15 , and 12 rats survived for 8 h in the control , 3 mg/kg , and 10 mg/kg groups , respectively . The control group had the lowest blood pressure , pulse rate , pH and arterial oxygen tension , and the highest arterial carbon dioxide tension and plasma IL-1beta concentration . The increase of P01375 was significantly lower in 3 mg/kg group than in the control group . Pathological changes of the lung were inhibited in 3 and 10 mg/kg groups . In conclusion , sarpogrelate might be effective to decrease production of pro-inflammatory cytokines , to keep hemodynamics , to inhibit lung damage , and to decrease mortality in endotoxin shock . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Clinical utility of the oral JAK inhibitor tofacitinib in the treatment of rheumatoid arthritis . Immune/inflammatory cells act in rheumatoid arthritis ( RA ) -affected patients by synthesizing several inflammatory mediators , including cytokines that initiate intracellular signaling . Recently , small molecule inhibitors of transduction and transcription signals that influence the intracellular pathways ( such as the Janus kinase [ JAK ] family of tyrosine kinases ) have been tested for RA treatment . Four members of the JAK family are known : P23458 , O60674 , P52333 , and TyK2 . P23458 / P52333 constitutively binds to the cytoplasmic portion of the cytokine receptor - the common gamma chain - that represents a common subunit of several cytokines involved in T-cell and natural killer cell development , as well as in B-cell activation . DB08895 is an oral JAK inhibitor that is now available and effective in RA treatment , as shown in multiple Phase II and Phase III clinical trials . However , long-term safety data and comparisons with other disease-modifying antirheumatic drugs and small molecule inhibitors are necessary to better determine the role of tofacitinib in RA . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . The deubiquitinating enzyme Q9Y5K5 interacts with Smads and regulates TGF-beta signalling . Disruption of components in the transforming growth factor-beta ( TGF-beta ) signalling cascade is a common occurrence in human cancers . TGF-beta pathway activation is accomplished via serine/threonine kinase receptors and intracellular Smad transcription factors . A key regulatory step involves specific ubiquitination by Smurfs that mediate the proteasomal degradation of Smads and/or receptors . Here , we report a novel interaction between Smads and ubiquitin C-terminal hydrolase Q9Y5K5 , a deubiquitinating enzyme that could potentially reverse Smurf-mediated ubiquitination . In Q86UG4 pull down experiments , Q9Y5K5 bound weakly to Q15796 and P84022 , and bound very strongly to O15105 in a region that is distinct from the -PY- motif in O15105 that interacts with Smurf ubiquitin ligases . Endogenous O15105 and Q9Y5K5 formed a stable complex in U4A/ P23458 cells , and FLAG- O15105 co-immunoprecipitated with HA- Q9Y5K5 in transfected P29320 -293 cells . In addition , we show that Q9Y5K5 can deubiquitinate and stabilize the type I TGF-beta receptor . Furthermore , overexpression of Q9Y5K5 upregulates TGF-beta-dependent transcription , and this effect is reversed in cells subject to RNAi-mediated knockdown of endogenous Q9Y5K5 . These findings support a new role for deubiquitinating enzymes in the control of the TGF-beta signalling pathway , and provide a novel molecular target for the design of inhibitors with therapeutic potential in cancer . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . N-arachidonoyl-L-serine is neuroprotective after traumatic brain injury by reducing apoptosis . N-arachidonoyl-L-serine ( AraS ) is a brain component structurally related to the endocannabinoid family . We investigated the neuroprotective effects of AraS following closed head injury induced by weight drop onto the exposed fronto-parietal skull and the mechanisms involved . A single injection of AraS following injury led to a significant improvement in functional outcome , and to reduced edema and lesion volume compared with vehicle . Specific antagonists to CB2 receptors , transient receptor potential vanilloid 1 ( Q8NER1 ) or large conductance calcium-activated potassium ( BK ) channels reversed these effects . Specific binding assays did not indicate binding of AraS to the Q9Y2T6 cannabinoid receptor . N-arachidonoyl-L-serine blocked the attenuation in phosphorylated extracellular-signal-regulated kinase 1/2 ( P29323 ) levels and led to an increase in pAkt in both the ipsilateral and contralateral cortices . Increased levels of the prosurvival factor Bcl-xL were evident 24 hours after injury in AraS-treated mice , followed by a 30 % reduction in caspase-3 activity , measured 3 days after injury . Treatment with a CB2 antagonist , but not with a P21554 antagonist , reversed this effect . Our results suggest that administration of AraS leads to neuroprotection via P29323 and Akt phosphorylation and induction of their downstream antiapoptotic pathways . These protective effects are related mostly to indirect signaling via the CB2R and Q8NER1 channels but not through P21554 or Q9Y2T6 receptors . Cannabinoid receptor activation induces apoptosis through tumor necrosis factor alpha-mediated ceramide de novo synthesis in colon cancer cells . PURPOSE : Cannabinoids have been recently proposed as a new family of potential antitumor agents . The present study was undertaken to investigate the expression of the two cannabinoid receptors , P21554 and CB2 , in colorectal cancer and to provide new insight into the molecular pathways underlying the apoptotic activity induced by their activation . EXPERIMENTAL DESIGN : Cannabinoid receptor expression was investigated in both human cancer specimens and in the DLD-1 and HT29 colon cancer cell lines . The effects of the P21554 agonist arachinodyl-2'-chloroethylamide and the CB2 agonist N-cyclopentyl-7-methyl-1-(2-morpholin-4-ylethyl)-1,8-naphthyridin-4(1H)-on-3-carboxamide ( CB13 ) on tumor cell apoptosis and ceramide and tumor necrosis factor ( P01375 ) -alpha production were evaluated . The knockdown of P01375 mRNA was obtained with the use of selective small interfering RNA . RESULTS : We show that the P21554 receptor was mainly expressed in human normal colonic epithelium whereas tumor tissue was strongly positive for the CB2 receptor . The activation of the P21554 and , more efficiently , of the CB2 receptors induced apoptosis and increased ceramide levels in the DLD-1 and HT29 cells . Apoptosis was prevented by the pharmacologic inhibition of ceramide de novo synthesis . The CB2 agonist CB13 also reduced the growth of DLD-1 cells in a mouse model of colon cancer . The knockdown of P01375 mRNA abrogated the ceramide increase and , therefore , the apoptotic effect induced by cannabinoid receptor activation . CONCLUSIONS : The present study shows that either P21554 or CB2 receptor activation induces apoptosis through ceramide de novo synthesis in colon cancer cells . Our data unveiled , for the first time , that P01375 acts as a link between cannabinoid receptor activation and ceramide production . 15d-PGJ2 and rosiglitazone suppress Janus kinase- P35610 inflammatory signaling through induction of suppressor of cytokine signaling 1 ( O15524 ) and O14543 in glia . Peroxisome proliferator-activated receptor ( Q07869 ) -gamma agonists are now emerging as therapeutic drugs for various inflammatory diseases . However , their molecular mechanism of action remains to be elucidated . Here we report a novel mechanism that underlies the P37231 agonist-mediated suppression of brain inflammation . We show that 15-deoxy-Delta12,14-prostaglandin J(2) ( 15d-PGJ(2) ) and rosiglitazone reduce the phosphorylation of P42224 and P40763 as well as P23458 ( P23458 ) and O60674 in activated astrocytes and microglia . The P37231 agonist-mediated reduction in phosphorylation leads to the suppression of JAK- P35610 -dependent inflammatory responses . The effects of 15d-PGJ(2) and rosiglitazone are not mediated by activation of P37231 . 15d-PGJ(2) and rosiglitazone rapidly induce the transcription of suppressor of cytokine signaling ( Q9NSE2 ) 1 and 3 , which in turn inhibit JAK activity in activated glial cells . In addition , Src homology 2 domain-containing protein phosphatase 2 ( SHP2 ) , another negative regulator of JAK activity , is also involved in their anti-inflammatory action . Our data suggest that 15d-PGJ(2) and rosiglitazone suppress the initiation of JAK- P35610 inflammatory signaling independently of P37231 , thus attenuating brain inflammation . Requirement of histone deacetylase activity for signaling by P42224 . P42224 is a transcription factor that plays a crucial role in signaling by interferons ( IFNs ) . In this study we demonstrated that inhibitors of histone deacetylase ( HDAC ) activity , butyrate , trichostatin A , and suberoylanilide hydroxamic acid , prevented IFNgamma-induced P23458 activation , P42224 phosphorylation , its nuclear translocation , and P42224 -dependent gene activation . Furthermore , we showed that silencing of Q13547 , Q92769 , and O15379 through RNA interference markedly decreased IFNgamma-driven gene activation and that overexpression of Q13547 , Q92769 , and O15379 enhanced P42224 -dependent transcriptional activity . Our data therefore established the essential role of deacetylase activity in P42224 signaling . Induction of P10914 by IFNgamma requires functional P42224 signaling and was abrogated by butyrate , trichostatin A , suberoylanilide hydroxamic acid , and P42224 small interfering RNA . In contrast , silencing of P42224 did not interfere with IFNgamma-induced expression of P52630 and caspase-7 , and HDAC inhibitors did not preclude IFNgamma-induced expression of P42224 , P52630 , and caspase-7 , suggesting that HDAC inhibitors impede the expression of IFNgamma target genes whose expression depends on P42224 but do not interfere with P42224 -independent signaling by IFNgamma . Finally , we showed that inhibitors of deacetylase activity sensitized colon cancer cells to IFNgamma-induced apoptosis through cooperative negative regulation of Bcl-x expression , demonstrating that interruption of the balance between P42224 -dependent and P42224 -independent signaling significantly alters the biological activity of IFNgamma . Novel treatments with small molecules in psoriatic arthritis . Current treatment options for patients with active psoriatic arthritis ( PsA ) include synthetic disease-modifying antirheumatic drugs and biologic agents . Propelled by increased understanding of immunopathogenesis of PsA , new therapeutic agents targeting different biologic pathways have been evaluated . This article discusses novel small-molecule , orally available treatments that are currently in clinical development for the treatment of psoriasis and PsA . This includes the phosphodiesterase 4 inhibitor apremilast and Janus kinase ( JAK ) inhibitors . Apremilast has demonstrated significant improvements in patients with moderate to severe psoriasis and PsA in phase II and III clinical trials and has recently been approved for the treatment of PsA . DB08895 , an oral inhibitor of P52333 , P23458 , and , to a lesser degree , O60674 , approved for the treatment of rheumatoid arthritis in several countries , has demonstrated positive results in psoriasis in phase II studies . Studies in PsA are ongoing . With these new developments , treatment options will continue to improve in the future . Tofacitinab in renal transplantation . DB08895 ( tositinib , CP-690,550 ) is a small molecule inhibitor of Janus associated kinases , primarily P52333 and O60674 , which inhibits cytokine signaling through the IL-2Rγ chain . In this article , we review the mechanism of action of tofacitinib , and pre-clinical and clinical data regarding its use in solid organ transplantation thus far . It is hoped that tofacitinib may form the basis for calcineurin-free immunosuppression , improving renal function while eliminating calcineurin inhibitor renal toxicity . Current studies suggest that tofacitinib is an effective immunosuppressive agent for renal transplantation , but it 's use in current protocols carries an increased risk of CMV , BK , and EBV viral infection , anemia and leukopenia , and post-transplant lymphoproliferative disorder . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . P10912 targeting to lipid rafts requires extracellular subdomain 2 . P10912 ( P10912 ) is a single membrane-spanning glycoprotein dimer that binds GH in its extracellular domain ( O95905 ) . GH activates the P10912 intracellular domain ( ICD ) -associated tyrosine kinase , O60674 , which causes intracellular signaling . We previously found that plasma membrane ( PM ) -associated P10912 was dramatically enriched in the lipid raft ( LR ) component of the membrane and that localization of P10912 within PM regions may regulate GH signaling by influencing the profile of pathway activation . In this study , we examined determinants of LR localization of the P10912 using a reconstitution system which lacks endogenous O60674 and P10912 . By non-detergent extraction and multistep fractionation , we found that P10912 was highly enriched in the LR fraction independent of O60674 expression . Various P10912 mutants were examined in transfectants harboring O60674 . LR concentration was observed for a P10912 in which the native transmembrane domain ( TMD ) is replaced by that of the unrelated P01130 and for a P10912 that lacks its ICD . Thus , LR association requires neither the TMD nor the ICD . Similarly , a P10912 that lacks the O95905 , except for the membrane-proximal O95905 stem region , was only minimally LR-concentrated . Mutants with internal stem deletions in the context of the full-length receptor were LR-concentrated similar to the wild-type . A P10912 lacking O95905 subdomain 1 reached the PM and was LR-concentrated , while one lacking O95905 subdomain 2 , also reached the PM , but was not LR-concentrated . These data suggest LR targeting resides in O95905 subdomain 2 , a region relatively uninvolved in GH binding . Concentrations of pravastatin and lovastatin in cerebrospinal fluid in healthy subjects . The capability of pravastatin and lovastatin , P04035 inhibitors likely to be taken chronically for hypercholesterolemia , to cross the blood-brain barrier was investigated in normal male volunteers . DB00227 , which is lipophilic , was detected in cerebrospinal fluid ( P04141 ) at concentrations that may have a pharmacologic effect . DB00175 , which is hydrophilic , was not detected in P04141 . It is concluded that pravastatin may have less potential for causing CNS-related side effects than lovastatin . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . Kinase inhibitors : a new class of antirheumatic drugs . The outlook for patients with rheumatoid arthritis has improved significantly over the last three decades with the use of disease-modifying antirheumatic drugs . However , despite the use of methotrexate , cytokine inhibitors , and molecules targeting T and B cells , a percentage of patients do not respond or lose their response over time . The autoimmune process in rheumatoid arthritis depends on activation of immune cells , which utilize intracellular kinases to respond to external stimuli such as cytokines , immune complexes , and antigens . In the past decade , small molecules targeting several kinases , such as p38 MAPK , Syk , and JAK have been developed . Several p38 MAPK inhibitors proved ineffective in treating rheumatoid arthritis . The Syk inhibitor , fostamatinib , proved superior to placebo in Phase II trials and is currently under Phase III investigation . DB08895 , a P23458 /3 inhibitor , was shown to be efficacious in two Phase III trials , while VX-509 , a P52333 inhibitor , showed promising results in a Phase II trial . Fostamatinib and tofacitinib were associated with increased rates of infection , elevation of liver enzymes , and neutropenia . Moreover , fostamatinib caused elevations of blood pressure and diarrhea , while tofacitinib was associated with an increase in creatinine and elevation of lipid levels . Pressure overload-induced cardiac hypertrophy response requires janus kinase 2-histone deacetylase 2 signaling . Pressure overload induces cardiac hypertrophy through activation of O60674 ( Jak2 ) , however , the underlying mechanisms remain largely unknown . In the current study , we tested whether histone deacetylase 2 ( Q92769 ) was involved in the process . We found that angiotensin II ( Ang-II ) -induced re-expression of fetal genes ( Atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) ) in cultured cardiomyocytes was prevented by the Jak2 inhibitor AG-490 and Q92769 inhibitor Trichostatin-A ( P32119 ) , or by Jak2/ Q92769 siRNA knockdown . On the other hand , myocardial cells with Jak2 or Q92769 over-expression were hyper-sensitive to Ang-II . In vivo , pressure overload by transverse aorta binding ( AB ) induced a significant cardiac hypertrophic response as well as re-expression of P01160 and DB04899 in mice heart , which were markedly reduced by AG-490 and P32119 . Significantly , AG-490 , the Jak2 inhibitor , largely suppressed pressure overload-/Ang-II-induced Q92769 nuclear exportation in vivo and in vitro . Meanwhile , P32119 or Q92769 siRNA knockdown reduced Ang-II-induced P01160 / DB04899 expression in Jak2 over-expressed H9c2 cardiomyocytes . Together , these results suggest that Q92769 might be a downstream effector of Jak2 to mediate cardiac hypertrophic response by pressure overload or Ang-II . The JAK inhibitor , tofacitinib , reduces the T cell stimulatory capacity of human monocyte-derived dendritic cells . OBJECTIVE : DB08895 , which is a Janus kinase ( JAK ) inhibitor , has shown clinical effects in the treatment of rheumatoid arthritis . JAKs are important kinases in lymphocyte differentiation ; however , their function in dendritic cells ( DCs ) is unknown . In this study , the function of JAKs in DCs was investigated with tofacitinib . METHODS : The effects of tofacitinib on the maturation of human monocyte-derived DCs induced by lipopolysaccharide ( LPS ) stimulation were investigated . In addition , its effects on T cell stimulatory capability was investigated by coculturing with naïve CD45RA-positive T cells . RESULTS : DB08895 decreased expression of P33681 / P42081 in a concentration-dependent manner in LPS-stimulated DCs ; however , it did not affect HLA-DR expression . DB08895 suppressed tumour necrosis factor , interleukin ( IL ) -6 and IL-1β production without affecting transforming growth factor ( TGF ) -β and P22301 production . Meanwhile , P33681 / P42081 expression in DCs was enhanced by type I interferon ( IFN ) stimulation , and the LPS-induced P33681 / P42081 expression was inhibited by an antibody to type I IFN receptor . Furthermore , tofacitinib suppressed production of type I IFN and activation of interferon regulatory factor ( Q969Q1 ) -7 , which is a transcription factor involved in P33681 / P42081 and type I IFN expression . DB08895 also decreased the T cell stimulatory capability of DCs and increased expression of indoleamine 2,3-dioxygenase ( P14902 ) -1 and Q6ZQW0 . CONCLUSIONS : DB08895 , a P23458 / P52333 inhibitor , affected the activities of human DCs . It decreased P33681 / P42081 expression and T cell stimulatory capability through suppression of type I IFN signalling . These results suggest a novel mode of action for tofacitinib and a pivotal role for JAKs in the differentiation of DCs . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Efficacy and safety of tofacitinib for treatment of rheumatoid arthritis . DB08895 is the first in a new class of nonbiologic disease-modifying antirheumatic drugs ( DMARDs ) , a targeted , synthetic DMARD , approved for the treatment of rheumatoid arthritis ( RA ) as monotherapy or in combination with methotrexate or other non-biologic DMARD . DB08895 , an orally administered Janus kinase ( JAK ) inhibitor , decreases T-cell activation , pro-inflammatory cytokine production , and cytokine signaling by inhibiting binding of type I cytokine receptors family and γ-chain cytokines to paired P23458 / P52333 receptors . The net effect of tofacitinb 's mechanism of action is decreased synovial inflammation and structural joint damage in RA patients . To date , six phase 3 trials have been conducted to evaluate the safety and efficacy of tofacitinib under the oral rheumatoid arthritis triaLs ( ORAL ) series . This review describes the pharmacology of the novel agent , tofacitinib , and details the safety and efficacy data of the ORAL trials . Multiple Gi proteins participate in nerve growth factor-induced activation of c-Jun N-terminal kinases in PC12 cells . Nerve growth factor ( P01138 ) -mediated activation of mitogen-activated protein kinases ( MAPK ) is critical for differentiation and apoptosis of PC12 cells . Since P01138 employs stress-activated c-Jun N-terminal kinase ( JNK ) to regulate both programmed cell death and neurite outgrowth of PC12 cells , we examined P01138 -regulated JNK activity and the role of G(i/o) proteins . Induction of JNK phosphorylation by P01138 occurred in a time- and dose-dependent manner and was partially inhibited by pertussis toxin ( PTX ) . To discern the participation of various signaling intermediates , PC12 cells were treated with specific inhibitors prior to P01138 challenge . P01138 -elevated JNK activity was abolished by inhibitors of JNK , p38 MAPK , Src , P52333 and Q02750 /2 . P01138 -dependent JNK phosphorylation became insensitive to PTX treatment upon transient expressions of Galpha(z) or the PTX-resistant mutants of Galpha(i1-3) and Galpha(oA) . Collectively , these studies indicate that P01138 -dependent JNK activity may be mediated via G(i1-3) proteins , P52333 , Src , p38 MAPK and the MEK/ P29323 cascade . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . Three lysine residues in the common β chain of the interleukin-5 receptor are required for Janus kinase ( JAK ) -dependent receptor ubiquitination , endocytosis , and signaling . Eosinophils are multifunctional leukocytes implicated in the pathogenesis of numerous inflammatory diseases including allergic asthma and hypereosinophilic syndrome . Eosinophil physiology is critically dependent on P05113 and the P05113 receptor ( IL-5R ) , composed of a ligand binding α chain ( IL-5Rα ) , and a common β chain , βc . Previously , we demonstrated that the βc cytoplasmic tail is ubiquitinated and degraded by proteasomes following P05113 stimulation . However , a complete understanding of the role of βc ubiquitination in IL-5R biology is currently lacking . By using a well established , stably transduced HEK293 cell model system , we show here that in the absence of ubiquitination , βc subcellular localization , P05113 -induced endocytosis , turnover , and IL-5R signaling were significantly impaired . Whereas ubiquitinated IL-5Rs internalized into trafficking endosomes for their degradation , ubiquitination-deficient IL-5Rs accumulated on the cell surface and displayed blunted signaling even after P05113 stimulation . Importantly , we identified a cluster of three membrane-proximal βc lysine residues ( Lys(457) , Lys(461) , and Lys(467) ) whose presence was required for both P23458 /2 binding to βc and receptor ubiquitination . These findings establish that JAK kinase binding to βc requires the presence of three critical βc lysine residues , and this binding event is essential for receptor ubiquitination , endocytosis , and signaling . Dysfunction of the P19957 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia . Patients with myeloproliferative disorders ( MPDs ) , such as essential thrombocythemia ( ET ) have increased risk of thrombosis and bleeding , which are major sources of morbidity and mortality . Most P53602 patients have a gain of function mutation in O60674 ( JAK2V617F ) , but little is known how JAK2V617F affects platelet function . Here , we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin ( which couples to O60674 ) on this pathway . In contrast , SFLLRN-mediated P16109 expression , DB00171 secretion , phosphorylation of the PKC substrate pleckstrin , and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets . In addition , thrombopoietin-mediated O60674 phosphorylation was unchanged , suggesting that signaling pathways activated downstream of O60674 are impaired . Indeed , we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the P19957 kinase substrate Akt , and have reduced activation of Rap1 in response to thrombopoietin , DB01277 ,ADP , SFLLRN , and thrombin . This effect was independent of Giα Q9H244 purinergic receptor function as ADP-mediated inhibition of P50552 phosphorylation was unchanged . These results demonstrate that the P19957 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients , resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation . Evidence for a role of phosphatidylinositol 3-kinase in P05112 -induced germline C epsilon transcription . Association of interleukin-4 receptor ( IL-4R ) with phosphatidylinositol 3-kinase ( P19957 -kinase ) has been demonstrated as the proximal event of P05112 signaling . We investigated the role of this enzyme in the P05112 signaling pathway in a human Burkitt lymphoma B cell line , DND39 , that expresses germline C epsilon transcripts in response to P05112 . Stimulation of DND39 cells with P05112 resulted in an accumulation of P19957 -monophosphate as well as a decrease of P29622 ,5-bisphosphate , which were abrogated by wortmannin , a potent inhibitor of P19957 -kinase . Activation of P19957 -kinase was further confirmed by the finding that P05112 caused an increase in P19957 -kinase activity coimmunoprecipitated with anti-IL-4R and with anti- P52333 kinase antibodies . As a possible downstream event of P19957 -kinase activation , the translocation of a zeta isoform of protein kinase C ( PKC ) from the cytosol to the membrane fraction was observed after P05112 stimulation , and wortmannin also suppressed this translocation . Moreover , P05112 -induced expression of germline C epsilon transcription was inhibited not only by wortmannin , but also by a PKC inhibitor , K252a . These results suggest that the signaling pathway involving P19957 -kinase and PKC zeta plays an important role in induction of germline C epsilon transcription in DND39 cells by P05112 . Activation of the JAK/ P35610 pathway in vascular smooth muscle by serotonin . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a vasoconstrictor and mitogen whose levels are elevated in diabetes . Previous studies have shown the presence of 5- Q13049 , P41595 , and P28222 receptors in vascular smooth muscle cells ( VSMCs ) . There are currently no data regarding P41595 and P28222 receptor activation of the JAK/ P35610 pathway in VSMCs and resultant potential alterations in 5-HT signaling in diabetes . Therefore , we tested the hypothesis that 5-HT differentially activates the JAK/ P35610 pathway in VSMCs under conditions of normal ( 5 mM ) and high ( 25 mM ) glucose . Treatment of rat VSMCs with 5-HT ( 10(-6) M ) resulted in time-dependent activation ( approximately 2-fold ) of O60674 , P23458 , and P42224 , but not P40763 ( maximal at 5 min , returned to baseline by 30 min ) . The P41595 receptor agonist BW723C86 and the P28222 receptor agonist CGS12066A ( 10(-9)-10(-5) M , 5-min stimulation ) did not activate the JAK/ P35610 pathway . Treatment with the 5- Q13049 receptor antagonist ketanserin ( 10 nM ) inhibited O60674 activation by 5-HT . Treatment of streptozotocin-induced diabetic rats with ketanserin ( 5 mg.kg-1.day-1 ) reduced activation of O60674 and P42224 but not P40763 in endothelium-denuded thoracic aorta in vivo . 5-HT ( 10(-6) M ) treatment resulted in increased cell proliferation and increased DNA synthesis , which were inhibited by the O60674 inhibitor AG490 . Further studies with apocynin , diphenyleneiodonium chloride , catalase , and virally transfected superoxide dismutase had no effect at either glucose concentration on activation of the JAK/ P35610 pathway by 5-HT . Therefore , we conclude that 5-HT activates O60674 , P23458 , and P42224 via the 5- Q13049 receptors in a reactive oxygen species-independent manner under both normal and high glucose conditions . Mixed adenoneuroendocrine carcinomas of the gastrointestinal tract : targeted next-generation sequencing suggests a monoclonal origin of the two components . BACKGROUND : Mixed adenoneuroendocrine carcinomas ( MANECs ) of the gastrointestinal tract are rare neoplasms characterized by coexisting exocrine and neuroendocrine neoplastic components . MANECs ' histogenetic classification and molecular characterization remain unclear , significantly affecting the identification of innovative therapeutic options for these tumors . METHODS : The exocrine and neuroendocrine components of 6 gastrointestinal MANECs were microdissected and subjected to the simultaneous mutation assessment in selected regions of 54 cancer-associated genes using Ion Torrent semiconductor-based next-generation sequencing . Sanger sequencing and immunohistochemistry were used as validation of the mutational status . RESULTS : A total of 20 driver gene somatic mutations were observed among the 12 neoplastic components investigated . In 11 of 12 ( 91.7 % ) samples , at least one mutation was detected ; 7 samples ( 58.3 % ) were found to have multiple mutations . P04637 gene mutations were the most frequent genetic alterations observed in the series , occurring in 11/12 samples ( 91.7 % ) . Somatic mutations in other genes were detected at lower frequencies : Q13315 , P35222 , Q15303 , P52333 , P35968 , P01116 , P06400 . CONCLUSIONS : Five of the 6 MANECs presented an overlapping mutational profile in both components , suggesting a monoclonal origin of the two MANEC components . Microcytosis in ank/ank mice and the role of Q9HCJ1 in promoting erythroid differentiation . Progressive ankylosis ( Ank and the human homolog , Q9HCJ1 ) is a transmembrane protein which regulates transport of inorganic pyrophosphate (PPi). ank/ank mice with a mutated ank gene , have calcification and bone ankylosis of the affected joints . In the course of studying these mutant mice , we found that they have microcytosis . These mutant mice have lower mean red blood cell volume ( MCV ) and lower hemoglobin content in red cells ( mean corpuscular hemoglobin , P20382 ) than normal mice . Using quantitative real-time PCR analysis , we showed that Ank was expressed in the E/Meg bipotent precursor , BFU-E , CFU-E , but there was no Ank expression in the hemoglobinizing erythroblasts . Stable Q9HCJ1 transfectants in K562 cells highly expressed two immature erythroid cell markers , P12830 and endoglin . Enhanced Erythropoietin ( Epo ) expression and downregulation of Q15466 -1 were detected in these transfectants . Consequently , the autocrine Epo-EpoR signaling pathway was activated , as evidenced by higher p- DB00135 O60674 , p- DB00135 EpoR and p- DB00135 P51692 in the Q9HCJ1 transfectants . Our results revealed a novel function of Q9HCJ1 in the promotion of early erythroid differentiation in K562 cells . We also showed that ank/ank mice have lower serum levels of Epo than the normal littermates , and this is the likely cause of microcytosis in these mutant mice . Preclinical to clinical translation of tofacitinib , a Janus kinase inhibitor , in rheumatoid arthritis . A critical piece in the translation of preclinical studies to clinical trials is the determination of dosing regimens that allow maximum therapeutic benefit with minimum toxicity . The preclinical pharmacokinetic ( PK ) /pharmacodynamic ( PD ) profile of tofacitinib , an oral Janus kinase ( JAK ) inhibitor , in a mouse collagen-induced arthritis ( mCIA ) model was compared with clinical PK/PD data from patients with rheumatoid arthritis ( RA ) . Preclinical evaluations included target modulation and PK/PD modeling based on continuous subcutaneous infusion or oral once- or twice-daily ( P55957 ) dosing paradigms in mice . The human PK/PD profile was obtained from pooled data from four phase 2 studies in patients with RA , and maximal effect models were used to evaluate efficacy after 12 weeks of tofacitinib treatment ( 1-15 mg P55957 ) . In mCIA , the main driver of efficacy was inhibition of cytokine receptor signaling mediated by P23458 heterodimers , but not O60674 homodimers , and continuous daily inhibition was not required to maintain efficacy . Projected efficacy could be predicted from total daily exposure irrespective of the oral dosing paradigm , with a total steady-state plasma concentration achieving 50 % of the maximal response ( Cave50 ) of ~100 nM . DB08895 potency ( ED50 ) in clinical studies was ~3.5 mg P55957 ( 90 % confidence interval : 2.3 , 5.5 ) or total Cave50 of ~40 nM , derived using Disease Activity Scores from patients with RA . The collective clinical and preclinical data indicated the importance of Cave as a driver of efficacy , rather than maximum or minimum plasma concentration ( Cmax or Cmin ) , where Cave50 values were within ~2-fold of each other . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Association of cytokeratin 7 and 19 expression with genomic stability and favorable prognosis in clear cell renal cell cancer . The purpose of our study was to demonstrate that distinct cytogenetic alterations in the most common subtype of renal cell cancer , clear cell renal cell carcinoma ( ccRCC ) , are reflected in protein expression profiles . We performed conventional cytogenetics and immunohistochemical analysis for cytokeratins ( CKs ) on 126 ccRCCs . Protein expression was evaluated in situ using a semiautomated quantitative system . The results were validated using an independent cohort of 209 ccRCCs with long-term follow-up . Cytogenetic alterations were identified in 96 of 126 ccRCCs , most of them involving chromosome 3 through loss , deletion or translocation . Expression of CKs and P12830 in ccRCC was associated with lack of cytogenetic alterations and low nuclear grade . In the validation set , CK7 and CK19 protein expression was associated with better clinical outcome . At the multivariate level , the best model included metastatic status and CK19 expression . Expression microarray analysis on 21 primary ccRCCs and 14 ccRCC metastases identified genes significantly associated with CK7 and CK19 expressing ccRCCs . Two novel ccRCC biomarkers associated with the CK7 positive ccRCC phenotype , P54278 and P50281 ( P50281 ) , were further validated . We conclude that the variability observed for CK expression in ccRCC can be explained by genetic heterogeneity . Distinct molecular subtypes of ccRCC with prognostic relevance were identified , and the CK7/CK19 expressing subtype is associated with better outcome . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . A pharmacokinetic and clinical assessment of tofacitinib for the treatment of rheumatoid arthritis . INTRODUCTION : Rheumatoid arthritis ( RA ) is a chronic painful and debilitating autoimmune disease . Although the outcome for patients with RA has improved markedly in the past decades , driven largely by the advent of biological disease-modifying antirheumatic drugs ( DMARDs ) and updated management strategies , adequate disease control can not be achieved in a substantial proportion of patients . Since RA is a syndrome with different biological subsets , DMARDs , with a novel mechanism of action , may represent a valuable addition to the current armamentarium . DB08895 is a novel synthetic DMARD that selectively inhibits Janus kinases ( JAKs ) , particularly P23458 and P52333 . AREAS COVERED : This review describes the pharmacokinetics of tofacitinib . Furthermore , the article summarizes and comments the drug 's efficacy and safety profile in RA patients . The authors furthermore assess data derived from the FDA 's RA development program . EXPERT OPINION : DB08895 is an oral synthetic DMARD displaying linear pharmacokinetics . Metabolism , primarily mediated by P08684 , accounts for 70 % of the total clearance of the drug ; the remaining 30 % are renally excreted . DB08895 monotherapy , or in combination with traditional DMARDs , has demonstrated its efficacy while having an acceptable safety profile in RA patients who have responded inadequately to current DMARDs , including P01375 antagonists . In view of its undetermined benefit to risk ratio , in the real-world population , tofacitinib should , for now , only be prescribed to selected patients . DB00588 -induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD1 and RFD7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 ) production . FP reduced the number of mature cells with a D1+ antigen-presenting phenotype and up-regulated the development of cells with the D1/D7+ and D7+ phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 ) . DB00588 also reversed the increase in both D1+ expression and P01375 production induced by P05112 . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Targeting P52333 in kidney transplantation : current status and future options . PURPOSE OF REVIEW : This review will discuss the mechanism of action and important clinical trial data in renal transplantation for the small molecule Janus kinase ( JAK ) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib . RECENT FINDINGS : JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors , particularly members of the cytokine receptor common gamma ( cγ ) chain family . P52333 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis . Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared with vehicle control . Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival . There was also a lower rate of new-onset diabetes after transplant . However , there was a trend toward more infections , including cytomegalovirus and BK virus nephritis . SUMMARY : DB08895 may be a promising alternative to calcineurin inhibitors . The optimal therapeutic window is still being determined . Activation of the JAK- P35610 pathway is necessary for desensitization of 5- Q13049 receptor-stimulated phospholipase C signalling by olanzapine , clozapine and MDL 100907 . We have previously demonstrated that olanzapine-induced desensitization of 5- Q13049 receptor-stimulated phospholipase C ( P98160 ) activity is associated with increases in P49802 protein levels both in vivo and in cells in culture , and the increase in P49802 is dependent on activation of the JAK- P35610 pathway in cells in culture . In the present study , we found that desensitization of 5- Q13049 receptor-stimulated P98160 activity induced by olanzapine is dependent on activation of the JAK- P35610 pathway . Similar to olanzapine , clozapine-induced desensitization of 5- Q13049 receptor signalling is accompanied by increases in P49802 and activation of O60674 . Treatment with the selective 5- Q13049 receptor antagonist MDL 100907 also increased P49802 protein levels and O60674 activation . Using a O60674 inhibitor AG490 , we found that clozapine and MDL 100907-induced increases in P49802 are dependent on activation of the JAK- P35610 pathway . Olanzapine , clozapine , and MDL 100907 treatment increased mRNA levels of P49802 . Using a chromatin immunoprecipitation assay we found P40763 binding to the putative P49802 promoter region . Taken together , olanzapine-induced activation of the JAK- P35610 pathway , and P40763 binding to the P49802 gene could underlie the increase in P49802 mRNA which could subsequently increase protein expression . Furthermore , the increase in P49802 protein could play a role in the desensitization of 5- Q13049 receptor signalling by terminating the activated Galphaq/11 proteins more rapidly . Overall , our data suggest that the complete desensitization of 5- Q13049 receptor-stimulated P98160 activity by olanzapine , clozapine and MDL 100907 requires activation of the JAK- P35610 pathway , which in turn increases P49802 expression probably by direct transcriptional activity of P40763 . Selective JAK/ P40763 signalling regulates transcription of colony stimulating factor-2 and -3 in Concanavalin-A-activated mesenchymal stromal cells . Human bone marrow-derived mesenchymal stromal cells ( MSCs ) express Toll-like receptors ( TLRs ) and produce cytokines and chemokines , all of which contribute to these cells ' immunomodulatory and proangiogenic properties . Among the secreted cytokines , colony-stimulating factors ( CSFs ) regulate angiogenesis through activation of endothelial cell proliferation and migration . Since O60682 are recruited within hypoxic tumors where they signal paracrine-regulated angiogenesis , the aim of this study was to evaluate which P04141 members are expressed and are inducible in activated O60682 . Furthermore , we investigated the JAK/ P35610 signal transducing pathway that may impact on P04141 transcription . O60682 were activated with Concanavalin-A ( ConA ) , a TLR-2/6 agonist as well as a membrane type-1 matrix metalloproteinase ( P50281 ) inducer , and we found increased transcription of granulocyte macrophage- P04141 ( GM- P04141 , P04141 -2 ) , granulocyte P04141 ( DB00099 , P04141 -3 ) , and P50281 . Gene silencing of either P40763 or P50281 prevented ConA-induced phosphorylation of P40763 , and reversed ConA effects on P04141 -2 and P04141 -3 . Treatment with the Janus Kinase (JAK)2 inhibitor AG490 antagonized the ConA induction of P50281 and P04141 -2 , while the pan-JAK inhibitor DB08895 reversed ConA-induced P04141 -2 and -3 gene expression . Silencing of O60674 prevented the ConA-mediated increase of P04141 -2 , while silencing of P23458 , P52333 and P29597 prevented the increase in P04141 -3 . Given that combined TLR-activation and locally-produced P04141 -2 and P04141 -3 could regulate immunomodulation and neovascularization , pharmacological targeting of TLR-2/6-induced P50281 /JAK/ P40763 signalling pathway may prevent O60682 contribution to tumor development . The effects of inhaled budesonide on circulating eosinophil progenitors and their expression of cytokines after allergen challenge in subjects with atopic asthma . Allergen inhalation by dual responder subjects with atopic asthma is associated with an increase in circulating eosinophil/basophil colony-forming units ( Eo/B CFU ) and granulocyte-macrophage colony- stimulating factor ( GM- P04141 ) immunolocalization in Eo/B colony cells grown in vitro . The current study examined the effect of the inhaled corticosteroid , budesonide , on the number of allergen- induced circulating eosinophils and Eo/B CFU , and immunolocalization of GM- P04141 and interleukin-5 ( P05113 ) in Eo/B colony cells grown in vitro . Sixteen subjects with mild atopic asthma were treated for either 7 or 8 d with 200 microg inhaled budesonide or placebo twice a day . Peripheral blood was collected before and 24 h after allergen inhalation challenge and nonadherent mononuclear cells ( NAMC ) were grown in methylcellulose culture . Eo/B CFU were enumerated after 14 d in culture , and prepared on slides for immunocytochemistry . DB01222 attenuated the allergen-induced increase in circulating eosinophils ( 4.0 +/- 0.4 x 10(5)/ml versus 6.5 +/- 0.7 x 10(5)/ml , p = 0.0001 ) , circulating Eo/B CFU ( 12.4 +/- 2.3/10(6) NAMC versus 18.8 +/- 4.6/10(6) NAMC , p = 0.05 ) , and immunolocalization of GM- P04141 in Eo/B colony cells ( 11.8 +/- 1.9 % positive versus 18.0 +/- 2.2 % , p = 0.01 ) but not immunolocalization of P05113 ( 7.9 +/- 1.4 % versus 4.5 +/- 0.6 % , p > 0.05 ) . Inhaled budesonide attenuated the number of allergen-induced circulating eosinophils and their progenitors grown in the presence of GM- P04141 , which may partially be a result of regulating eosinophil progenitor expression of the autocrine growth factor GM- P04141 . P25116 genotype influences platelet aggregation and procoagulant responses in patients with coronary artery disease prior to and during clopidogrel therapy . Genetic variations of the protease-activated receptor-1 ( P25116 ) have been associated with platelet receptor density and linked to thrombin receptor-activating peptide ( TRAP ) -induced phenotypes of platelet aggregation and P16109 expression . We investigated whether the P25116 intervening sequence-14 A > T dimorphism influences platelet procoagulant activity . We also determined whether the Q9H244 antagonist clopidogrel could offset any observed functional polymorphism of the P25116 receptor by inhibiting Q9H244 -mediated amplification of TRAP-induced responses . We studied 54 patients listed for elective percutaneous coronary intervention assessing TRAP-induced platelet aggregation and markers of procoagulant activity . Platelet responses were measured at baseline , 4 h post clopidogrel 300 mg , and 10 and 28 days following clopidogrel 75 mg daily . Each patient was genotyped for the P25116 intervening sequence-14 A/T dimorphism . Increased platelet aggregation and procoagulant responses were observed with P25116 A allele homozygotes . DB00758 significantly inhibited these platelet responses regardless of P25116 genotype , but did not offset the hyper-reactivity associated with the A/A homozygotes . We conclude that a common sequence variation within the P25116 gene influences TRAP-induced platelet procoagulant activity as well as aggregation . Higher platelet reactivity associated with P25116 IVSn-14 A allele homozygotes persists despite clopidogrel therapy . These individuals may be at higher risk of thromboembolic events and may require additional anti-platelet medication . Comparison of gene expression profiles and related pathways in chronic thromboembolic pulmonary hypertension . Chronic thromboembolic pulmonary hypertension ( CTEPH ) is one of the main causes of severe pulmonary hypertension . However , despite treatment ( pulmonary endarterectomy ) , in approximately 15-20 % of patients , pulmonary vascular resistance and pulmonary arterial pressure continue to increase . To date , little is known about the changes that occur in gene expression in CTEPH . The identification of genes associated with CTEPH may provide insight into the pathogenesis of CTEPH and may aid in diagnosis and treatment . In this study , we analyzed the gene expresion profiles of pulmonary artery endothelial cells from 5 patients with CTEPH and 5 healthy controls using oligonucleotide microarrays . Bioinformatics analyses using the Gene Ontology ( GO ) and KEGG databases were carried out to identify the genes and pathways specifically associated with CTEPH . Signal transduction networks were established to identify the core genes regulating the progression of CTEPH . A number of genes were found to be differentially expressed in the pulmonary artery endothelial cells from patients with CTEPH . In total , 412 GO terms and 113 pathways were found to be associated with our list of genes . All differential gene interactions in the Signal-Net network were analyzed . P52333 , P30679 , O15264 , P32121 and P25116 were the most significantly altered . Bioinformatics analysis may help gather and analyze large amounts of data in microarrays by means of rigorous experimental planning , scientific statistical analysis and the collection of complete data . In this study , a novel differential gene expression pattern was constructed . However , further studies are required to identify novel targets for the diagnosis and treatment of CTEPH . Concomitant loss of p120-catenin and β-catenin membrane expression and oral carcinoma progression with P12830 reduction . The binding of p120-catenin and β-catenin to the cytoplasmic domain of P12830 establishes epithelial cell-cell adhesion . Reduction and loss of catenin expression degrades P12830 -mediated carcinoma cell-cell adhesion and causes carcinomas to progress into aggressive states . Since both catenins are differentially regulated and play distinct roles when they dissociate from P12830 , evaluation of their expression , subcellular localization and the correlation with P12830 expression are important subjects . However , the same analyses are not readily performed on squamous cell carcinomas in which P12830 expression determines the disease progression . In the present study , we examined expression and subcellular localization of p120-catenin and β-catenin in oral carcinomas ( n = 67 ) and its implications in the carcinoma progression and P12830 expression using immunohitochemistry . At the invasive front , catenin-membrane-positive carcinoma cells were decreased in the dedifferentiated ( p120-catenin , P < 0.05 ; β-catenin , P < 0.05 ) and invasive carcinomas ( p120-catenin , P < 0.01 ; β-catenin , P < 0.05 ) and with the P12830 staining ( p120-catenin , P < 0.01 ; β-catenin , P < 0.01 ) . Carcinoma cells with β-catenin cytoplasmic and/or nuclear staining were increased at the invasive front compared to the center of tumors ( P < 0.01 ) . Although the p120-catenin isoform shift from three to one associates with carcinoma progression , it was not observed after TGF-β , P01133 or P01375 -α treatments . The total amount of p120-catenin expression was decreased upon co-treatment of TGF-β with P01133 or P01375 -α . The above data indicate that catenin membrane staining is a primary determinant for P12830 -mediated cell-cell adhesion and progression of oral carcinomas . Furthermore , it suggests that loss of p120-catenin expression and cytoplasmic localization of β-catenin fine-tune the carcinoma progression . EGCG enhances the therapeutic potential of gemcitabine and CP690550 by inhibiting P40763 signaling pathway in human pancreatic cancer . BACKGROUND : Signal Transducer and Activator of Transcription 3 ( P40763 ) is an oncogene , which promotes cell survival , proliferation , motility and progression in cancer cells . Targeting P40763 signaling may lead to the development of novel therapeutic approaches for human cancers . Here , we examined the effects of epigallocathechin gallate ( EGCG ) on P40763 signaling in pancreatic cancer cells , and assessed the therapeutic potential of EGCG with gemcitabine or P52333 inhibitor CP690550 ( DB08895 ) for the treatment and/or prevention of pancreatic cancer . METHODOLOGY/PRINCIPAL FINDINGS : Cell viability and apoptosis were measured by XTT assay and TUNEL staining , respectively . Gene and protein expressions were measured by qRT-PCR and Western blot analysis , respectively . The results revealed that EGCG inhibited the expression of phospho and total P52333 and P40763 , P40763 transcription and activation , and the expression of P40763 -regulated genes , resulting in the inhibition of cell motility , migration and invasion , and the induction of caspase-3 and PARP cleavage . The inhibition of P40763 enhanced the inhibitory effects of EGCG on cell motility and viability . Additionally , gemcitabine and CP690550 alone inhibited P40763 target genes and synergized with EGCG to inhibit cell viability and induce apoptosis in pancreatic cancer cells . CONCLUSIONS/SIGNIFICANCE : Overall , these results suggest that EGCG suppresses the growth , invasion and migration of pancreatic cancer cells , and induces apoptosis by interfering with the P40763 signaling pathway . Moreover , EGCG further enhanced the therapeutic potential of gemcitabine and CP690550 against pancreatic cancer . Janus kinase activation by cytokine oncostatin M decreases Q8NBP7 expression in liver cells . Q8NBP7 degrades P01130 ( P01130 ) in liver and thereby influences the circulating level of LDL cholesterol . Hence , mechanisms inhibiting Q8NBP7 expression have potential for cholesterol-lowering intervention . Previously , we demonstrated that oncostatin M ( OM ) activates P01130 gene transcription , resulting in an increased LDL uptake in HepG2 cells and a reduction of plasma LDL in hypercholesterolemic hamsters . Here we identify the suppression of Q8NBP7 expression by OM as one new mechanism that increases P01130 protein in HepG2 cells . Treating HepG2 cells with OM decreases Q8NBP7 mRNA and protein levels . Inhibition studies and small interfering RNA -targeted depletion revealed a critical role for P23458 and O60674 in mediating this OM inhibitory effect . Furthermore , we showed that OM induces transient phosphorylation of P42224 , P40763 , and P42229 and sustained activation of P29323 signaling molecules . While depletion of P35610 members in HepG2 cells did not affect OM inhibitory activity on Q8NBP7 expression , blocking activation of the Q02750 / P29323 signaling pathway resulted in attenuation of the OM inhibitory effect . Finally , by using an anti-hamster Q8NBP7 antibody , we demonstrated the in vivo suppression of liver Q8NBP7 mRNA and protein expression by OM in hypercholesterolemic hamsters . Our study uncovered a cytokine-triggered regulatory network for Q8NBP7 expression that is linked to JAKs and the P29323 signaling pathway .

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interacts_with DB01045?

multiple_choice

Heritable susceptibility factors for the development of cancer . High frequencies of inherited DNA sequence variations ( polymorphisms ) are found in the human population . The involvement of polymorphic genes ( especially for chemical metabolism and DNA repair ) in the development of cancer is under intensive investigation . In our studies , we have irradiated blood lymphocytes from normal non-smokers with gamma-rays or UV-light to investigate genotypes and DNA repair functions . We found that P18887 399Gln and O43542 241Met were deficient in the repair of gamma-ray-but not UV-light-induced DNA damage that led to the expression of chromosome aberrations ; therefore the variant genotypes are defective in base excision repair . The reverse was found with P18074 312Asn and P18074 751Gln ; therefore they are defective in nucleotide excision repair . P18887 194Trp , O15527 326Cys and P27695 148Glu had no DNA repair deficiency based on our experimental conditions . In another study , we investigated the role of some of these genes on the development of lung cancer . We found a significant increase of chromosome aberrations in patients and controls that had the P18074 751Gln and P09488 null genotypes , indicating a mechanistic causation of the disease . Therefore , inheritance of susceptibility genes can have significant impact on disease burden in the population . On the other hand , there are many questions that need to be addressed in order to evaluate the impact of susceptibility on cancer . These questions include the understanding of combinations of different polymorphic genes for susceptibility and of specific disease susceptibility for different ethnic populations . Further investigation of the effect of framework catenation on hydrogen uptake in metal-organic frameworks . Hydrogen-sorption studies have been carried out for the catenation isomer pairs of Q15149 -6 and Q15149 -6 ' ( both have the formula of Cu(3)(TATB)(2) , where TATB represents 4,4',4''-s-triazine-2,4,6-triyl-tribenzoate with a formula of C(24)H(12)N(3)O(6) ) . Inelastic neutron scattering ( P01308 ) studies reveal that the initial sites occupied by adsorbed H(2) are the open Cu centers of the paddlewheel units with comparable interaction energies in the two isomers . At high H(2) loadings , where the H(2) molecules adsorb mainly on or around the organic linkers , the interaction is found to be substantially stronger in catenated Q15149 -6 than in noncatenated Q15149 -6 ' , leading to much higher H(2) uptake in the isomer with catenation . Hydrogen sorption measurements at pressures up to 50 bar demonstrate that framework catenation can be favorable for the enhancement of hydrogen adsorption . For example , the excess hydrogen uptake of Q15149 -6 is 72 mg/g ( 6.7 wt % ) at 77 K/50 bar or 9.3 mg/g ( 0.92 wt % ) at 298 K/50 bar , respectively , and that for Q15149 -6 ' is 42 mg/g ( 4.0 wt % ) at 77 K/50 bar or 4.0 mg/g ( 0.40 wt % ) at 298 K/50 bar . Importantly , Q15149 -6 exhibits a total hydrogen uptake of 95 mg/g ( 8.7 wt % ) ( corresponding to a total volumetric value of 53.0 g/L , estimated based on crystallographic density ) at 77 K/50 bar and 15 mg/g ( 1.5 wt % ) at 298 K/50 bar . Significantly , the expected usable capacity of Q15149 -6 is as high as 75 mg/g ( or 41.9 g/L ) at 77 K , if a recharging pressure of 1.5 bar is assumed . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . DB01045 Does not Significantly Affect the Expression of Small Heterodimer Partner in Primary Human Hepatocytes . The small/short heterodimer partner ( Q15466 , Q15466 ) is a nuclear receptor corepressor lacking a DNA binding domain . Q15466 is induced by bile acid-activated farnesoid X receptor ( Q96RI1 ) resulting in P22680 gene suppression . In contrast , O75469 ( O75469 ) activation by its ligands was recently suggested to inhibit Q15466 gene transactivation to maximize the induction of O75469 target genes . However , there are also conflicting reports in literature whether O75469 or rodent Pxr activation down-regulates Q15466 /Shp expression . Moreover , the O75469 -mediated regulation of the Q15466 gene has been studied only at the Q15466 mRNA and transactivation ( gene reporter assay ) levels . In this study , we studied the effect of rifampicin , a prototype O75469 ligand , on Q15466 mRNA , and protein expression in three primary human hepatocyte cultures . We found that Q15466 mRNA is not systematically down-regulated in hepatocyte in culture after 24 h treatment with rifampicin . Consistently , we did not observe down-regulation of Q15466 protein in primary human hepatocytes after 24 and 48 h of incubation with rifampicin . We can conclude that although we observed slight down-regulation of Q15466 mRNA and protein in several hepatocyte preparations , the phenomenon is unlikely critical for O75469 -mediated induction of its target genes . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Dimethylarsinic acid in drinking water changed the morphology of urinary bladder but not the expression of DNA repair genes of bladder transitional epithelium in F344 rats . Inorganic arsenic increases urinary bladder transitional cell carcinoma in humans . In F344 rats , dimethylarsinic acid ( P28067 [V] ) increases transitional cell carcinoma . Arsenic-induced inhibition of DNA repair has been reported in cultured cell lines and in lymphocytes of arsenic-exposed humans , but it has not been studied in urinary bladder . Should inhibition of DNA damage repair in transitional epithelium occur , it may contribute to carcinogenesis or cocarcinogenesis . We investigated morphology and expression of DNA repair genes in F344 rat transitional cells following up to 100 ppm P28067 (V) in drinking water for four weeks . Mitochondria were very sensitive to P28067 (V) , and swollen mitochondria appeared to be the main source of vacuoles in the transitional epithelium . Real-time reverse transcriptase polymerase chain reaction ( Real-Time RT PCR ) showed the mRNA levels of tested DNA repair genes , ataxia telangectasia mutant ( Q13315 ) , X-ray repair cross-complementing group 1 ( P18887 ) , excision repair cross-complementing group 3/xeroderma pigmentosum B ( P19447 / P19447 ) , and P06746 ( Polbeta ) , were not altered by P28067 (V) . These data suggested that either P28067 (V) does not affect DNA repair in the bladder or P28067 (V) affects DNA repair without affecting baseline mRNA levels of repair genes . The possibility remains that P28067 (V) may lower damage-induced increases in repair gene expression or cause post-translational modification of repair enzymes . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Effects of DB01045 , a potent inducer of drug-metabolizing enzymes and an inhibitor of Q9Y6L6 /3 transport , on the single dose pharmacokinetics of anacetrapib . Anacetrapib is a novel cholesteryl ester transfer protein ( P11597 ) inhibitor in development for treatment of dyslipidemia . This open-label , fixed-sequence , 3-period study was intended to evaluate the potential of anacetrapib to be a victim of Q9Y6L6 /3 inhibition and strong CYP3A induction using acute and chronic dosing of rifampin , respectively , as a probe . In this study , 16 healthy subjects received 100 mg anacetrapib administered without rifampin ( Day 1 , Period 1 ) , with single-dose ( SD ) 600 mg rifampin ( Day 1 , Period 2 ) , and with multiple-dose ( MD ) 600 mg rifampin for 20 days ( Day 14 , Period 3 ) . Log-transformed anacetrapib AUC0-∞ and Cmax were analyzed by a linear mixed effects model . The GMRs and 90 % CIs for anacetrapib AUC0-∞ and Cmax were 1.25 ( 1.04 , 1.51 ) and 1.43 ( 1.13 , 1.82 ) for SD rifampin ( Period 2/Period 1 ) and 0.35 ( 0.29 , 0.42 ) and 0.26 ( 0.21 , 0.32 ) for MD rifampin ( Period 3/Period 1 ) , respectively . Anacetrapib was generally well tolerated in both the absence/presence of SD and MD rifampin . In conclusion , treatment with SD rifampin , which inhibits the Q9Y6L6 /3 transporter system , did not substantially influence the SD pharmacokinetics of anacetrapib , while chronic ( 20 days ) administration of rifampin , which strongly induces CYP3A isozymes , reduced mean systemic exposure to SD anacetrapib by 65 % . Attenuation of anti-tuberculosis therapy induced hepatotoxicity by Spirulina fusiformis , a candidate food supplement . Therapy using Isoniazid ( DB00951 ) and DB01045 ( Q9HBH0 ) leads to induction of hepatotoxicity in some individuals undergoing anti-tuberculosis treatment . In this study , we assessed the effect of Spirulina fusiformis on DB00951 and Q9HBH0 induced hepatotoxicity in rats compared with hepatoprotective drug Silymarin . Induction of hepatotoxicity was measured by changes in the liver marker enzymes ( aspartate transaminase , alanine transaminase , and alkaline phosphatase ) . The antioxidant status was also analyzed in liver tissue homogenate and plasma by measurement of superoxide dismutase , catalase , glutathione-S-transferase , glutathione reductase , and lipid peroxidation levels . We also aimed to study the binding and interactions of the transcription factors Pregnane X Receptor ( O75469 ) and Farnesoid X Receptor ( Q96RI1 ) with DB00951 , Q9HBH0 , and representative active compounds of Spirulina fusiformis by in silico methods . The administration of DB00951 and Q9HBH0 resulted in significant ( p < 0.05 ) decrease in the antioxidant levels and total protein levels . There was also a significant ( p < 0.05 ) increase in the levels of liver marker enzymes . Spirulina fusiformis was seen to protect the parameters from significant changes upon challenge with DB00951 and Q9HBH0 in a dose-dependent manner . This was corroborated by histological examination of the liver . The results of the in silico analyses further support the wet lab results . Gelatin nanocarriers as potential vectors for effective management of tuberculosis . The aim of the research work was to develop and characterize rifampicin ( Q9HBH0 ) loaded gelatin nanoparticulate delivery system for the effective management of tuberculosis . Gelatin nanoparticles ( GPs ) containing Q9HBH0 were prepared using two-step desolvation method . Formulations were characterized through transmission electron microscopy ( TEM ) , atomic force microscopy ( P43652 ) , size and size distribution analysis , polydispersity index ( P07237 ) , zeta potential , percent drug entrapment , percent nanoparticulate yield and in vitro drug release . Formulations were further characterized for in vitro cytotoxicity , in vivo biodistribution , and antitubercular activity . The nanoparticles were found to be spherical in shape . The size of nanoparticles was found to be 264+/-11.2 nm with low P07237 suggesting the narrow particle size distribution . The drug release showed the biphasic pattern of release i.e. initial burst followed by a sustained release pattern . The cytotoxicity studies revealed that nanoparticles are safe , non toxic as compared to free drug . In vivo biodistribution study showed higher localization of Q9HBH0 loaded GPs in various organs , as compared to plain Q9HBH0 solution in PBS ( pH 7.4 ) . In contrast to free drug , the nanoparticles not only sustained the plasma level but also enhanced the AUC and mean residence time ( MRT ) of the drug , suggesting improved pharmacokinetics of drug . Q9HBH0 GPs additionally resulted in significant reduction in bacterial counts in the lungs and spleen of TB-infected mice . Hence , GPs hold promising potential for increasing drug targetability vis a vis reducing dosing frequency with the interception of minimal side effects , for efficient management of tuberculosis . P29474 activation by HDL is impaired in genetic P11597 deficiency . Mutations in the P11597 gene resulting in defective P11597 activity have been shown to cause remarkable elevations of plasma HDL-C levels , with the accumulation in plasma of large , buoyant HDL particles enriched in apolipoprotein E. Genetic P11597 deficiency thus represents a unique tool to evaluate how structural alterations of HDL impact on HDL atheroprotective functions . Aim of the present study was to assess the ability of HDL obtained from P11597 -deficient subjects to protect endothelial cells from the development of endothelial dysfunction . HDL isolated from one homozygous and seven heterozygous carriers of P11597 null mutations were evaluated for their ability to down-regulate cytokine-induced cell adhesion molecule expression and to promote NO production in cultured endothelial cells . When compared at the same protein concentration , HDL and HDL3 from carriers proved to be as effective as control HDL and HDL3 in down-regulating cytokine-induced P19320 , while carrier HDL2 were more effective than control HDL2 in inhibiting P19320 expression . On the other hand , HDL and HDL fractions from carriers of P11597 deficiency were significantly less effective than control HDL and HDL fractions in stimulating NO production , due to a reduced P29474 activating capacity , likely because of a reduced Q14703 content . In conclusion , the present findings support the notion that genetic P11597 deficiency , by affecting HDL particle structure , impacts on HDL vasculoprotective functions . Understanding of these effects might be important for predicting the outcomes of pharmacological P11597 inhibition . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Pregnane X Receptor Represses HNF4α Gene to Induce P01308 -Like Growth Factor-Binding Protein P08833 that Alters Morphology of and Migrates HepG2 Cells . Upon treatment with the pregnane X receptor ( O75469 ) activator rifampicin ( Q9HBH0 ) , human hepatocellular carcinoma HepG2-derived ShP51 cells that stably express O75469 showed epithelial-mesenchymal transition ( EMT ) -like morphological changes and migration . Our recent DNA microarrays have identified hepatocyte nuclear factor ( HNF ) 4α and insulin-like growth factor-binding protein ( IGFBP ) 1 mRNAs to be downregulated and upregulated , respectively , in Q9HBH0 -treated ShP51 cells , and these regulations were confirmed by the subsequent real-time polymerase chain reaction and Western blot analyses . Using this cell system , we demonstrated here that the O75469 -HNF4α- P08833 pathway is an essential signal for O75469 -induced morphological changes and migration . First , we characterized the molecular mechanism underlying the O75469 -mediated repression of the HNF4α gene . Chromatin conformation capture and chromatin immunoprecipitation ( ChIP ) assays revealed that O75469 activation by Q9HBH0 disrupted enhancer-promoter communication and prompted deacetylation of histone H3 in the HNF4α P1 promoter . Cell-based reporter and ChIP assays showed that O75469 targeted the distal enhancer of the HNF4α P1 promoter and stimulated dissociation of HNF3β from the distal enhancer . Subsequently , small interfering RNA knockdown of HNF4α connected O75469 -mediated gene regulation with the O75469 -induced cellular responses , showing that the knockdown resulted in the upregulation of P08833 and EMT-like morphological changes without Q9HBH0 treatment . Moreover , recombinant P08833 augmented migration , whereas an anti- P08833 antibody attenuated both O75469 -induced morphological changes and migration in ShP51 cells . O75469 indirectly activated the P08833 gene by repressing the HNF4α gene , thus enabling upregulation of P08833 to change the morphology of ShP51 cells and cause migration . These results provide new insights into O75469 -mediated cellular responses toward xenobiotics including therapeutics . The mechanism of the G0/ P55008 cell cycle phase arrest induced by activation of O75469 in human cells . CONTEXT : O75469 ( O75469 ) is an important transcriptional regulator that plays important roles in the cell metabolism and cell growth by regulating the transcriptional of a sort of metabolizing enzymes . OBJECTIVE : To investigate whether rifampicin effected HepG2 cells growth and the inhibition was due to the G0/ P55008 phase arrest . METHODS : O75469 -knockdown experiments using RNAi showed that the cell cycle phase arrest mediated by rifampicin based on activation of O75469 . The results also indicated that cell phase arrest by rifampicin could protect cells form UVB-induced DNA damage . P19793 ( RXRα ) expression level in cells is another key factor for cell cycle phase arrest mediated by rifampicin . Both over expression and lacking expression of RXRα in cell reduced the cell arrest efficiency mediated by rifampicin . In the study , we found that rifampicin inhibited HepG2 cells growth and demonstrated that the inhibition is due to the G0/ P55008 phase arrest through flow cytometry analysis . CONCLUSION : The results showed that RXRα promote cell cycle phase transition rate of HepG2 . Competitive bind of rifampicin-activated O75469 with RXRα is one main reason to arrest cell cycle phase through inhibiting combination of RXRα with other partners . DB01045 could promote cell growth rate when RXRα expressed more excessively than O75469 in cells . Evidence of drug-drug interactions through uptake and efflux transport systems in rat hepatocytes : implications for cellular concentrations of competing drugs . For drugs with hepatobiliary transport across hepatocytes , the interplay between uptake and efflux transporters determines hepatic concentrations of drugs , but the evolution over time of these concentrations is difficult to measure in humans other than with magnetic resonance imaging contrast agents in the liver . DB00743 dimeglumine ( BOPTA ) is a contrast agent used in liver magnetic resonance imaging that enters into human hepatocytes through organic anion transporting polypeptides ( P46721 ) and exits unchanged into bile through the multiple resistance-associated protein 2 ( Q92887 ) . DB01045 ( Q9HBH0 ) is transported by the same membrane proteins and may compete with BOPTA for hepatic uptake . Simultaneous drug-drug interactions through uptake and efflux transport systems in hepatocytes according to the cellular concentrations of competing drugs were never investigated . In perfused rat liver preparations , we demonstrate how the drug-drug interactions through transporters determine cellular concentrations of the competing drugs BOPTA and Q9HBH0 , and we show that the cellular concentrations by modulating transport through membranes regulate the rat Oatp-Mrp2 interplay . Moreover , drug interactions through transporters change greatly over time . Dose-dependent modulation of apoptotic processes by fluoxetine in maturing neuronal cells : an in vitro study . OBJECTIVES : Recent studies indicate that the selective serotonin reuptake inhibitor ( SSRI ) fluoxetine is not solely effective by the instant inhibition of the serotonin transporter ( P31645 ) but also by its influence on mitotic and/or apoptotic processes . METHODS : To investigate the effects of the compound in vitro , we treated neurons from different brain areas with increasing concentrations of fluoxetine . Additionally , human embryonic kidney ( P29320 -293 ) cells and P29320 -293 cells stably expressing the P31645 were used . Cell viability was quantified by MTT-assay and apoptosis via fluorescence-activated cell-sorting analyses . DB00472 's effect on the γ-aminobutyric acid ( GABA ) receptor was electrophysiologically investigated to test the hypothesis if a GABA-mimetic effect exists that might lead - additionally to the well-known N-methyl-D-aspartate ( DB01221 ) -antagonism - to increased apoptosis in immature neurons . RESULTS : In hippocampal , cortical , and both types of P29320 -293 cells , viability decreased and apoptosis increased in a dose-dependent manner ( 0.5-75 μM ) . In contrast , in mesencephalic and striatal cells the viability was unchanged or even slightly stimulated up to 20 μM fluoxetine . An anti-apoptotic effect of concentrations below 10 μM was observed in these cells . The GABA(A) receptor was directly activated by fluoxetine . CONCLUSIONS : We conclude that fluoxetine affects apoptotic processes independently from P31645 expression . Since especially the combined GABA-mimetic and DB01221 -antagonistic effects increase apoptosis in developing neuronal cells , whereas both effects are neuroprotective in adult neurons we hypothesise that these mechanisms explain the discrepancy of in vitro and in vivo studies . 24S-hydroxycholesterol induces cholesterol release from choroid plexus epithelial cells in an apical- and apoE isoform-dependent manner concomitantly with the induction of O95477 and P45844 expression . The release of cholesterol from choroid plexus epithelial cells ( P16870 ) plays an important role in cholesterol homeostasis in the P04141 . The purpose of this study was to clarify the molecules involved in cholesterol release in P16870 and the regulation mechanisms of the cholesterol release by the liver X receptor ( LXR ) using a conditionally immortalized P16870 line ( TR-CSFB3 ) . The mRNA expression of LXRalpha , LXRbeta and their target genes , DB00171 -binding cassette transporter (ABC)A1 , P45844 , Q9H172 and Q9H222 , were detected in rat choroid plexus . O95477 and P45844 protein were detected in the plasma membrane of TR-CSFB3 cells . Following treatment with 24S-hydroxycholesterol , an endogenous LXR ligand , the expression of O95477 and P45844 were induced in TR-CSFB3 cells . Moreover , apolipoprotein (apo)AI- and high-density lipoprotein ( HDL ) -mediated cholesterol release to the apical side of TR-CSFB3 cells was facilitated by this treatment , whereas that to the basal side was not affected . Following 24S-hydroxycholesterol treatment , apoE3-dependent cholesterol release from TR-CSFB3 cells was enhanced more than the apoE4-dependent release . These results suggest that LXR activation facilitates cholesterol release into the P04141 from P16870 through the functional induction of O95477 and P45844 . The difference between apoE3 and apoE4 suggests that the cholesterol release from P16870 is related to the development of neurodegenerative diseases . O75469 induces Q02318 and regulates cholesterol metabolism in the intestine . Mitochondrial sterol 27-hydroxylase ( Q02318 ) catalyzes oxidative cleavage of the sterol side chain in the bile acid biosynthetic pathway in the liver and 27-hydroxylation of cholesterol in most tissues . Recent studies suggest that 27-hydroxycholesterol ( 27-HOC ) activates liver orphan receptor alpha ( LXRalpha ) and induces the cholesterol efflux transporters O95477 and P45844 in macrophages . The steroid- and bile acid-activated pregnane X receptor ( O75469 ) plays critical roles in the detoxification of bile acids , cholesterol metabolites , and xenobiotics . The role of Q02318 in the intestine is not known . This study investigated O75469 and Q02318 regulation of cholesterol metabolism in the human intestinal cell lines Caco2 and Ls174T . A human O75469 ligand , rifampicin , induced Q02318 mRNA expression in intestine cells but not in liver cells . DB01045 induced Q02318 gene transcription , increased intracellular 27-HOC levels , and induced O95477 and P45844 mRNA expression only in intestine cells . A functional O75469 binding site was identified in the human Q02318 gene . Chromatin immunoprecipitation assays revealed that rifampicin induced the O75469 recruitment of steroid receptor coactivator 1 to Q02318 chromatin . DB04540 loading markedly increased intracellular 27-HOC levels in intestine cells . DB01045 , 27-HOC , and a potent LXRalpha agonist , T0901317 , induced O95477 and P45844 protein expression and stimulated cholesterol efflux from intestine cells to apolipoprotein A-I and HDL . This study suggests an intestine-specific O75469 / Q02318 /LXRalpha pathway that regulates intestine cholesterol efflux and HDL assembly . A structural and in vitro characterization of asoprisnil : a selective progesterone receptor modulator . Selective progesterone receptor modulators ( SPRMs ) have been suggested as therapeutic agents for treatment of gynecological disorders . One such SPRM , asoprisnil , was recently in clinical trials for treatment of uterine fibroids and endometriosis . We present the crystal structures of progesterone receptor ( PR ) ligand binding domain complexed with asoprisnil and the corepressors nuclear receptor corepressor ( NCoR ) and Q9Y618 . This is the first report of steroid nuclear receptor crystal structures with ligand and corepressors . These structures show PR in a different conformation than PR complexed with progesterone ( P4 ) . We profiled asoprisnil in PR-dependent assays to understand further the PR-mediated mechanism of action . We confirmed previous findings that asoprisnil demonstrated antagonism , but not agonism , in a PR-B transfection assay and the T47D breast cancer cell alkaline phosphatase activity assay . Asoprisnil , but not DB00834 , weakly recruited the coactivators Q15788 and Q9Y6Q9 . However , asoprisnil strongly recruited the corepressor NCoR in a manner similar to DB00834 . Unlike DB00834 , NCoR binding to asoprisnil-bound PR could be displaced with equal affinity by NCoR or Q15596 peptides . We further showed that it weakly activated T47D cell gene expression of Sgk-1 and O60437 and antagonized P4-induced expression of both genes . In rat leiomyoma ELT3 cells , asoprisnil demonstrated partial P4-like inhibition of cyclooxygenase ( P36551 ) enzymatic activity and P35354 gene expression . In the rat uterotrophic assay , asoprisnil demonstrated no P4-like ability to oppose estrogen . Our data suggest that asoprisnil differentially recruits coactivators and corepressors compared to DB00834 or P4 , and this specific cofactor interaction profile is apparently insufficient to oppose estrogenic activity in rat uterus . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . Early detection of tumor response to chemotherapy by 3'-deoxy-3'-[18F]fluorothymidine positron emission tomography : the effect of cisplatin on a fibrosarcoma tumor model in vivo . We have assessed the potential of [18F]fluorothymidine positron emission tomography ( [18F] P17948 -PET ) to measure early cytostasis and cytotoxicity induced by cisplatin treatment of radiation-induced fibrosarcoma 1 ( Q9HBH0 -1 ) tumor-bearing mice . DB00515 -mediated arrest of tumor cell growth and induction of tumor shrinkage at 24 and 48 hours , respectively , were detectable by [18F] P17948 -PET . At 24 and 48 hours , the normalized uptake at 60 minutes ( tumor/liver radioactivity ratio at 60 minutes after radiotracer injection ; NUV60 ) for [18F] P17948 was 0.76 +/- 0.08 ( P = 0.03 ) and 0.51 +/- 0.08 ( P = 0.03 ) , respectively , compared with controls ( 1.02 +/- 0.12 ) . The decrease in [18F] P17948 uptake at 24 hours was associated with a decrease in cell proliferation assessed immunohistochemically ( a decrease in proliferating cell nuclear antigen labeling index , LI( P12004 ) , from 14.0 +/- 2.0 % to 6.2 +/- 1.0 % ; P = 0.001 ) , despite the lack of a change in tumor size . There were P55008 -S and G2-M phase arrests after cisplatin treatment , as determined by cell cycle analysis . For the quantitative measurement of tumor cell proliferation , [18F] P17948 -PET was found to be superior to [18F] DB09150 -PET ( NUV60 versus LIPCNA : r = 0.89 , P = 0.001 and r = 0.55 , P = 0.06 , respectively ) . At the biochemical level , we found that the changes in [18F] P17948 and [18F] DB09150 uptake were due to changes in levels of thymidine kinase 1 protein , hexokinase , and DB00171 . This work supports the further development of [18F] P17948 -PET as a generic pharmacodynamic readout for early quantitative imaging of drug-induced changes in cell proliferation in vivo . Effect of prototypical inducing agents on P-glycoprotein and CYP3A expression in mouse tissues . P-glycoprotein ( P-gp ) and CYP3A have considerable overlap in inducers in vitro . Characterizing P-gp induction in vivo and potential coregulation with CYP3A are important goals for predicting drug interactions . This study examined P-gp expression in mouse tissues and potential coinduction with CYP3A following oral treatment with 1 of 7 prototypical inducing agents for 5 days . P-gp expression in brain or liver was not induced by any treatment as determined by Western blot , whereas dexamethasone , pregnenolone-16alpha-carbonitrile ( Q15149 ) , St . John 's wort ( SJW ) , and rifampin induced hepatic CYP3A expression . In intestine , rifampin and SJW induced P-gp expression 3.7- and 1.6-fold and CYP3A 3.5- and 2.4-fold , respectively , whereas dexamethasone and Q15149 induced CYP3A only . These observations suggest that P-gp in mouse small intestine is inducible by some , but not all , CYP3A inducers , whereas P-gp expression in liver or brain is not readily induced . Intriguingly , rifampin and SJW , both activators of the human pregnane X receptor ( O75469 ) , induced CYP3A in both liver and intestine but induced P-gp only in intestine , whereas Q15149 , an activator of murine O75469 , did not induce P-gp in any tissue . DB01045 disposition was evaluated , and hepatic exposure to rifampin was comparable to intestine ; in contrast , brain concentrations were low . Overall , these observations demonstrate that P-gp induction in vivo is tissue-specific ; furthermore , there is a disconnect between P-gp induction and CYP3A induction that is tissue- and inducer-dependent , suggesting that O75469 activation alone is insufficient for P-gp induction in vivo . Tissue-specific factors and inducer pharmacokinetic/pharmacodynamic properties may underlie these observations . DB01045 -independent interactions between the pregnane X receptor ligand binding domain and peptide fragments of coactivator and corepressor proteins . The pregnane X receptor ( O75469 ) , a member of the nuclear receptor superfamily , regulates the expression of drug-metabolizing enzymes in a ligand-dependent manner . The conventional view of nuclear receptor action is that ligand binding enhances the receptor 's affinity for coactivator proteins , while decreasing its affinity for corepressors . To date , however , no known rigorous biophysical studies have been conducted to investigate the interaction among O75469 , its coregulators , and ligands . In this work , steady-state total internal reflection fluorescence microscopy ( TIRFM ) and total internal reflection with fluorescence recovery after photobleaching were used to measure the thermodynamics and kinetics of the interaction between the O75469 ligand binding domain and a peptide fragment of the steroid receptor coactivator-1 ( Q15788 ) in the presence and absence of the established O75469 agonist , rifampicin . Equilibrium dissociation and dissociation rate constants of ~5 μM and ~2 s(-1) , respectively , were obtained in the presence and absence of rifampicin , indicating that the ligand does not enhance the affinity of the O75469 and Q15788 fragments . Additionally , TIRFM was used to examine the interaction between O75469 and a peptide fragment of the corepressor protein , the silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) . An equilibrium dissociation constant of ~70 μM was obtained for Q9Y618 in the presence and absence of rifampicin . These results strongly suggest that the mechanism of ligand-dependent activation in O75469 differs significantly from that seen in many other nuclear receptors . Elevated retinol binding protein 4 induces apolipoprotein B production and associates with hypertriglyceridemia . CONTEXT AND OBJECTIVE : A high level of retinol binding protein 4 ( P02753 ) is reported to be associated with insulin resistance in humans . However , evidence from large-scale populations about the relationship between serum P02753 and metabolic phenotypes is scarce . In the present study , we aimed to evaluate serum P02753 distribution and its association with metabolic phenotypes among middle-aged and elderly Chinese . DESIGN AND PARTICIPANTS : Serum concentrations of P02753 in a cross-sectional sample of 2780 Chinese population aged 50-70 years old in Guangzhou were measured by ELISA . RESULTS : The mean of serum P02753 concentration was 28.04 μg/mL for male and 37.76 μg/mL for female ( P < .01 ) , respectively . Circulating P02753 was positively correlated with serum triglyceride and apolipoprotein B ( apoB ) concentrations . The odds ratio ( OR ) was substantially higher for hypertriglyceridemia ( OR , 3.26 ; 95 % confidence interval , 2.36-4.51 ) in the highest P02753 quartile compared with those in the lowest quartile after multiple adjustment for confounders . Furthermore , serum P02753 was significantly associated with fasting glucose , insulin levels , and homeostasis model assessment index-insulin resistance ( HOMA-IR ) . Moreover , we showed that P02753 enhanced microsomal triglyceride transfer protein ( P55157 ) expression and activity via up-regulation of protein disulfide isomerase ( P07237 ) , suppressed low-density lipoprotein receptor ( P01130 ) expression , and impaired insulin-signaling pathway , leading to inductions in apoB secretion both in vitro and in vivo . CONCLUSIONS : Elevated circulating P02753 concentrations were associated with higher risk of hypertriglyceridemia by inducing the secretion of triglyceride-rich apoB-containing lipoproteins . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . P12004 D1b represses breast cancer cell growth by antagonizing the action of cyclin D1a on estrogen receptor alpha-mediated transcription . Alternate splicing of the cyclin D1 gene gives rise to transcripts a and b which encode two protein isoforms cyclin D1a and cyclin D1b . P12004 D1a can substitute for estrogen to activate estrogen receptor alpha- ( ERalpha ) mediated transcription and can induce the proliferation of estrogen responsive tissues . However , little is known about the biological role of cyclin D1b in transcriptional regulation . In this study , we determined that cyclin D1b is incapable of inducing ERalpha-mediated transcription because it fails to recruit steroid receptor coactivator-1 ( Q15788 ) to ERalpha . Moreover , cyclin D1b antagonizes cyclin D1a-induced ERalpha-mediated transcription by competing with cyclin D1a for ERalpha binding . Cell proliferation assay showed that cyclin D1b repressed the ERalpha-positive breast cancer T47D cell growth . Our findings suggest that the cyclin D1b represses breast cancer cell growth by antagonizing the action of cyclin D1a on ERalpha-mediated transcription . Cytokine profile in the endometrium of normal fertile and women with repeated implantation failure . BACKGROUND : Repeated Implantation Failure ( Q9HBH0 ) is one of the most intricate obstacles in assisted reproduction . The cytokine and chemokine composition of uterine cavity seem to play important roles in the implantation process . OBJECTIVE : To compare the cytokine profile in the endometrium of normal fertile women and those with repeated implantation failure . METHODS : After enzymatic digestion of endometrial tissues , whole endometrial cells and endometrial stromal cells from Q9HBH0 and normal fertile women were cultivated and stimulated for cytokine secretion . The levels of P22301 , TGF-β , IFN-γ , P05231 , P10145 and Q16552 in culture supernatants of the two groups were assayed by ELISA and compared together . RESULTS : Endometrial stromal cells and whole endometrial cells of normal fertile women produced higher levels of P05231 , P10145 and TGF-β compared to Q9HBH0 group , although this difference was statistically significant only in endometrial stromal cells ( p=0.005 , 0.002 and 0.001 , respectively ) . In addition , endometrial stromal cells of normal fertile women produced lower levels of P22301 in comparison with Q9HBH0 group ( p=0.005 ) . CONCLUSION : Disturbances in cytokine production at the feto-maternal interface could be a cause of implantation failure . A pro-inflammatory cytokine milieu seems to be pivotal for successful implantation . The genetics of complex cholestatic disorders . Cholestatic liver diseases are caused by a range of hepatobiliary insults and involve complex interactions among environmental and genetic factors . Little is known about the pathogenic mechanisms of specific cholestatic diseases , which has limited our ability to manage patients with these disorders . However , recent genome-wide studies have provided insight into the pathogenesis of gallstones , primary biliary cirrhosis , and primary sclerosing cholangitis . A lithogenic variant in the gene that encodes the hepatobiliary transporter Q9H221 has been identified as a risk factor for gallstone disease ; this variant has been associated with altered cholesterol excretion and metabolism . Other variants of genes encoding transporters that affect the composition of bile have been associated with cholestasis , namely O95342 , which encodes the bile salt export pump , and P21439 , which encodes hepatocanalicular phosphatidylcholine floppase . In contrast , studies have associated primary biliary cirrhosis and primary sclerosing cholangitis with genes encoding major histocompatibility complex proteins and identified loci associated with microbial sensing and immune regulatory pathways outside this region , such as genes encoding IL12 , Q14765 , Q13568 , P60568 and its receptor ( IL2R ) , P10747 , and P33681 . These discoveries have raised interest in the development of reagents that target these gene products . We review recent findings from genetic studies of patients with cholestatic liver disease . Future characterization of genetic variants in animal models , stratification of risk alleles by clinical course , and identification of interacting environmental factors will increase our understanding of these complex cholestatic diseases . DB04540 metabolite , 5-cholesten-3β-25-diol-3-sulfate , promotes hepatic proliferation in mice . Oxysterols are well known as physiological ligands of liver X receptors ( LXRs ) . Oxysterols , 25-hydroxycholesterol ( 25HC ) and 27-hydroxycholesterol as endogenous ligands of LXRs , suppress cell proliferation via LXRs signaling pathway . Recent reports have shown that sulfated oxysterol , 5-cholesten-3β-25-diol-3-sulfate ( 25HC3S ) as LXRs antagonist , plays an opposite direction to oxysterols in lipid biosynthesis . The present report was to explore the effect and mechanism of 25HC3S on hepatic proliferation in vivo . Following administration , 25HC3S had a 48 h half life in the circulation and widely distributed in mouse tissues . Profiler™ PCR array and RTqPCR analysis showed that either exogenous or endogenous 25HC3S generated by overexpression of oxysterol sulfotransferase ( SULT2B1b ) plus administration of 25HC significantly up-regulated the proliferation gene expression of Wt1 , Pcna , cMyc , cyclin A , FoxM1b , and CDC25b in a dose-dependent manner in liver while substantially down-regulating the expression of cell cycle arrest gene Chek2 and apoptotic gene Apaf1 . Either exogenous or endogenous administration of 25HC3S significantly induced hepatic DNA replication as measured by immunostaining of the P12004 labeling index and was associated with reduction in expression of LXR response genes , such as O95477 and SREBP-1c . Synthetic LXR agonist T0901317 effectively blocked 25HC3S-induced hepatic proliferation . CONCLUSIONS : 25HC3S may be a potent regulator of hepatocyte proliferation and oxysterol sulfation may represent a novel regulatory pathway in liver proliferation via inactivating LXR signaling . Dietary calcium decreases plasma cholesterol by down-regulation of intestinal Niemann-Pick C1 like 1 and microsomal triacylglycerol transport protein and up-regulation of P22680 and ABCG 5/8 in hamsters . SCOPE : It has been shown that calcium supplementation favorably modifies plasma lipoprotein profile in postmenopausal women . The present study investigated the interaction of dietary calcium with genes of transporters , receptors and enzymes involved in cholesterol metabolism . METHODS AND RESULTS : Forty-eight ovariectomized hamsters were fed one of the four diets containing 0 , 2 , 6 and 8 g calcium per kg . Plasma total cholesterol ( TC ) , triacylglycerols ( TG ) , and non-high density lipoprotein cholesterol were dose-dependently decreased , whereas high-density lipoprotein cholesterol ( HDL-C ) was dose-dependently increased with the increasing dietary calcium levels . Dietary calcium had no effect on protein mass of hepatic sterol regulatory element binding protein-2 ( SREBP ) , liver X receptor-alpha ( LXR ) , 3-hydroxy-3-methylglutaryl- DB01992 reductase ( HMGR ) , P01130 ( P01130 ) and cholesterol-7α-hydroxylase ( P22680 ) . However , dietary calcium up-regulated the mRNA levels of hepatic P22680 and intestinal DB00171 binding cassette transporters ( Q9H222 /8 ) whereas it down-regulated the intestinal Niemann-Pick C1 like 1 ( Q9UHC9 ) and microsomal triacylglycerol transport protein ( P55157 ) . In addition , dietary calcium increased the activity of intestinal acyl coenzyme A : cholesterol acyltransferase 2 , while it decreased plasma cholesteryl ester transport protein ( P11597 ) . CONCLUSION : Beneficial modification of lipoprotein profile by dietary calcium was mediated by sequestering bile acid absorption and enhancing excretion of fecal cholesterol , via up-regulation of mRNA P22680 and intestinal ABCG 5/8 with down-regulation of mRNA Q9UHC9 and P55157 . Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism . BACKGROUND : Cardiovascular disease ( CVD ) is one of the major causes of death worldwide . Numerous genetic risk factors in lipid metabolism , including mutations of P01130 , P04114 , and Q8NBP7 , as well as polymorphisms of P11597 and P02649 , have been found to associate with CVD . METHODS : In this study , a two-dye based arrayed primer extension ( P27695 ) microarray assay for simultaneous multigene ( P01130 , P04114 , Q8NBP7 , P11597 , and P02649 ) detection was developed . The DNA templates , originating from 1 DNA sample of known genotype and 7 blind DNA samples , were amplified by uniplex PCR . RESULTS : Optimized conditions for the P27695 reaction were determined to include a hybridization temperature of 55°C and a DNA template size of 50-150bp . The total assay including PCR , purification , fragmentation , P27695 reaction , and image analysis could be performed in 6h . In total , 48 genotypes were identified among 8 individual DNA samples by P27695 analysis . CONCLUSIONS : The data suggest that this P27695 microarray offers a robust , fast , and versatile option for screening these genotypes in hypercholesterolemia patients . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . PLA(2) signaling is involved in calpain-mediated degradation of synaptic dihydropyrimidinase-like 3 protein in response to DB01221 excitotoxicity . Q14117 -like 3 ( Q14195 ) is believed to play a role in neuronal differentiation , axonal outgrowth and neuronal regeneration , as well as cytoskeleton organization . Recently we have shown that glutamate excitotoxicity and oxidative stress result in calpain-dependent cleavage of Q14195 , and that NOS plays a role in this process [ R . Kowara , Q . Chen , M. Milliken , B . Chakravarthy , Calpain-mediated truncation of dihydropyrimidinase-like 3 protein ( Q14195 ) in response to DB01221 and H2O2 toxicity , J. Neurochem. 95 ( 2005 ) 466-474 ; R. Kowara , K.L. Moraleja , B. Chakravarthy , Involvement of nitric oxide synthase and ROS-mediated activation of L-type voltage-gated Ca(2+) channels in DB01221 -induced Q14195 degradation , Brain Res . 1119 ( 2006 ) 40-49 ] . The present study investigates the involvement of PLA(2) signaling in DB01221 -induced Q14195 degradation . Exposure of rat primary cortical neurons ( Q15149 ) to PLA(2) and P35354 inhibitors significantly prevented DB01221 -induced Q14195 degradation . Since the metabolic product of PLA(2) signaling , PGE(2) , which augments toxic effect of DB01221 , is known to stimulate DB02527 , the effect of adenyl cyclase activator ( forskolin plus DB07954 ) and inhibitor ( MDL12,300 ) on DB01221 -induced Q14195 degradation was tested . Our data indicate that the activation of adenyl cyclase contributes to DB01221 -induced Q14195 degradation . Furthermore , DB02527 -dependent protein kinase ( PKA ) inhibitor PKI ( 14-22 ) provided additional evidence of PKA involvement in DB01221 -induced Q14195 degradation . In summary , the obtained data show the contribution of PLA(2) signaling to DB01221 -induced calpain activation and subsequent degradation of synaptic protein Q14195 . Synthesis of new thiazolo[4,5-d]pyrimidines as DB01285 releasing factor modulators . P06850 ( CRF ) is a neurohormone that plays a crucial role in integrating the body 's overall response to stress . It appears necessary and sufficient for the organism to mount functional , physiological and endocrine responses to stressors . CRF is released in response to various triggers such as chronic stress . The role of CRF and its involvement in these neurological disorders suggest that new drugs that can target the CRF function or bind to its receptors may represent a new development of neuropsychiatric medicines to treat various stress-related disorders including depression , anxiety and addictive disorders . Based on pharmacophore of the CRF1 receptor antagonists , a new series of thiazolo[4,5-d] pyrimidines were synthesized as P06850 ( CRF ) receptor modulators and the prepared compounds carry groups shown to produce optimum binding affinity to CRF receptors . Twenty two compounds were evaluated for their CRF1 receptor binding affinity in P29320 293 cell lines and two compounds 5o and 5s showed approximately 25 % binding affinity to CRF1 receptors . Selected compounds ( 5c and 5f ) were also evaluated for their effect on expression of genes associated with depression and anxiety disorders such as CRF1 , P16220 , P21397 , P31645 , P01303 , DatSLC6a3 , and P09172 and significant upregulation of CRF1 mRNA has been observed with compound 5c . Hinokitiol Negatively Regulates Immune Responses through Cell Cycle Arrest in Concanavalin A-Activated Lymphocytes . Autoimmune diseases are a group of chronic inflammatory diseases that arise from inappropriate inflammatory responses . Hinokitiol , isolated from the wood of Chamaecyparis taiwanensis , engages in multiple biological activities . Although hinokitiol has been reported to inhibit inflammation , its immunological regulation in lymphocytes remains incomplete . Thus , we determined the effects of hinokitiol on concanavalin A- ( ConA- ) stimulated T lymphocytes from the spleens of mice . In the present study , the MTT assay revealed that hinokitiol ( 1-5 μM ) alone did not affect cell viability of lymphocytes , but at the concentration of 5 μM it could reduce ConA-stimulated T lymphocyte proliferation . Moreover , propidium iodide ( PI ) staining revealed that hinokitiol arrested cell cycle of T lymphocytes at the G0/ P55008 phase . Hinokitiol also reduced interferon gamma ( IFN-γ ) secretion from ConA-activated T lymphocytes , as detected by an ELISA assay . In addition , hinokitiol also downregulated cyclin D3 , Q01094 , and Cdk4 expression and upregulated P38936 expression . These results revealed that hinokitiol may regulate immune responses . In conclusion , we for the first time demonstrated that hinokitiol upregulates P38936 expression and attenuates IFN-γ secretion in ConA-stimulated T lymphocytes , thereby arresting cell cycle at the G0/ P55008 phase . In addition , our findings also indicated that hinokitiol may provide benefits to treating patients with autoimmune diseases . An integrative in silico approach for discovering candidates for drug-targetable protein-protein interactions in interactome data . BACKGROUND : Protein-protein interactions ( PPIs ) are challenging but attractive targets for small chemical drugs . Whole PPIs , called the ' interactome ' , have been emerged in several organisms , including human , based on the recent development of high-throughput screening ( HTS ) technologies . Individual PPIs have been targeted by small drug-like chemicals ( SDCs ) , however , interactome data have not been fully utilized for exploring drug targets due to the lack of comprehensive methodology for utilizing these data . Here we propose an integrative in silico approach for discovering candidates for drug-targetable PPIs in interactome data . RESULTS : Our novel in silico screening system comprises three independent assessment procedures : i ) detection of protein domains responsible for PPIs , ii ) finding P18827 -binding pockets on protein surfaces , and iii ) evaluating similarities in the assignment of Gene Ontology ( GO ) terms between specific partner proteins . We discovered six candidates for drug-targetable PPIs by applying our in silico approach to original human PPI data composed of 770 binary interactions produced by our HTS yeast two-hybrid ( HTS-Y2H ) assays . Among them , we further examined two candidates , P19793 / P48552 and P24941 / P38936 , with respect to their biological roles , PPI network around each candidate , and tertiary structures of the interacting domains . CONCLUSION : An integrative in silico approach for discovering candidates for drug-targetable PPIs was applied to original human PPIs data . The system excludes false positive interactions and selects reliable PPIs as drug targets . Its effectiveness was demonstrated by the discovery of the six promising candidate target PPIs . Inhibition or stabilization of the two interactions may have potential therapeutic effects against human diseases . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . P51587 interacts with the cytoskeletal linker protein plectin to form a complex controlling centrosome localization . The breast cancer susceptibility gene ( P51587 ) is localized mainly in the nucleus where it plays an important role in DNA damage repair . Some P51587 protein is also present in the centrosome . Here , we demonstrate that P51587 interacts with plectin , a cytoskeletal cross-linker protein , and that this interaction controls the position of the centrosome . Phosphorylation of plectin by cyclin-dependent kinase 1/cyclin B ( P06493 /CycB ) kinase has been reported to abolish its cross-linking function during mitosis . Here , we induced phosphorylation of plectin in prepared fractions of HeLa cells by adding activated P06493 /CycB kinase . Consequently , there was significant dissociation of the centrosome from the nuclear membrane . Q15149 has six homologous ankyrin-like repeat domains ( termed Q15149 M1-M6 ) . Using a pull-down assay , we found that Q86UG4 - Q15149 M1 and a Q86UG4 -C-terminal region fusion protein ( which comprised Q15149 M6 , along with an adjacent vimentin site ) interacted with P51587 . Since each Q15149 module exhibits high homology to the others , the possibility of all six domains participating in this interaction was indicated . Moreover , when Q15149 M1 was overexpressed in HeLa cells , it competed with endogenous plectin and inhibited the P51587 -plectin interaction . This inhibitory effect resulted in dissociation of the centrosomes from the nucleus and increased the rate of micronuclei formation which may lead to carcinogenesis . In addition , when either P51587 or plectin was suppressed by the appropriate siRNA , a similar change in centrosomal positioning was observed . We suggest that the P51587 -plectin interaction plays an important role in the regulation of centrosome localization and also that displacement of the centrosome may result in genomic instability and cancer development . LPS-induced downregulation of Q92887 and O95342 in human liver is due to a posttranscriptional process . Endotoxin-induced cholestasis in rodents is caused by hepatic downregulation of transporters , including the basolateral Na+-dependent taurocholate transporter ( ntcp ) and the canalicular bile salt export pump ( bsep ) and multidrug resistance-associated protein 2 ( mrp2 ) . Details about the regulation of the human transporter proteins during this process are lacking . We used precision-cut human and rat liver slices to study the regulation of transporter expression during LPS-induced cholestasis . We investigated the effect of LPS on nitrate/nitrite and cytokine production in relation to the expression of inducible nitric oxide synthase , Q14973 , O95342 , and Q92887 both at the level of mRNA with RT-PCR and protein using immunofluorescence microscopy . In liver slices from both species , LPS-induced expression of inducible nitric oxide synthase was detected within 1-3 h and remained increased over 24 h . In rat liver slices , this was accompanied by a significant decrease of rat ntcp and mrp2 mRNA levels , whereas bsep levels were not affected . These results are in line with previous in vivo studies and validate our liver slice technique . In LPS-treated human liver slices , Q14973 mRNA was downregulated and showed an inverse correlation with the amounts of P01375 and Il-1beta produced . In contrast , Q92887 and O95342 mRNA levels were not affected under these conditions . However , after 24-h LPS challenge , both proteins were virtually absent in human liver slices , whereas marker proteins remained detectable . In conclusion , we show that posttranscriptional mechanisms play a more prominent role in LPS-induced regulation of human Q92887 and O95342 compared with the rat transporter proteins . NO-1886 suppresses diet-induced insulin resistance and cholesterol accumulation through P42229 -dependent upregulation of IGF1 and P22680 . P01308 resistance and dyslipidemia are both considered to be risk factors for metabolic syndrome . Low levels of IGF1 are associated with insulin resistance . Elevation of low-density lipoprotein cholesterol ( LDL-C ) concomitant with depression of high-density lipoprotein cholesterol ( HDL-C ) increase the risk of obesity and type 2 diabetes mellitus ( T2DM ) . Liver secretes IGF1 and catabolizes cholesterol regulated by the rate-limiting enzyme of bile acid synthesis from cholesterol 7alpha-hydroxylase ( P22680 ) . NO-1886 , a chemically synthesized lipoprotein lipase activator , suppresses diet-induced insulin resistance with the improvement of HDL-C . The goal of the present study is to evaluate whether NO-1886 upregulates IGF1 and P22680 to benefit glucose and cholesterol metabolism . By using human hepatoma cell lines ( HepG2 cells ) as an in vitro model , we found that NO-1886 promoted IGF1 secretion and P22680 expression through the activation of signal transducer and activator of transcription 5 ( P42229 ) . Pretreatment of cells with AG 490 , the inhibitor of P35610 pathway , completely abolished NO-1886-induced IGF1 secretion and P22680 expression . Studies performed in Chinese Bama minipigs pointed out an augmentation of plasma IGF1 elicited by a single dose administration of NO-1886 . Long-term supplementation with NO-1886 recovered hyperinsulinemia and low plasma levels of IGF1 suppressed LDL-C and facilitated reverse cholesterol transport by decreasing hepatic cholesterol accumulation through increasing P22680 expression in high-fat/high-sucrose/high-cholesterol diet minipigs . These findings indicate that NO-1886 upregulates IGF1 secretion and P22680 expression to improve insulin resistance and hepatic cholesterol accumulation , which may represent an alternative therapeutic avenue of NO-1886 for T2DM and metabolic syndrome . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Cloning , expression , and regulation of rabbit cyclooxygenase-2 in renal medullary interstitial cells . Prostaglandin synthesis requires cyclooxygenase-1 ( P23219 ) or -2 ( P35354 ) , which mediate the conversion of arachidonate to prostaglandin H2 . P23219 is the predominant constitutive isoform , whereas P35354 expression is typically low . In the present studies we cloned rabbit P35354 and determined its distribution in unstimulated tissues . Screening rabbit eye and uterine libraries yielded two cDNAs containing identical inserts with a 1,812-nucleotide open-reading frame . This encoded a 604-amino acid polypeptide , 90 % identical to human , rat , and mouse P35354 . Expression of the rabbit P35354 in P29320 -293 cells enhanced prostanoid synthesis . Constitutive P35354 mRNA expression was highest in kidney and urinary bladder . P35354 expression was primarily in renal outer medullary interstitial cells with cortical expression in macula densa . In cultured medullary interstitial cells , P35354 mRNA predominated , with little P23219 expression . Interstitial cell P35354 mRNA but not P23219 was induced by phorbol ester and epidermal growth factor but suppressed by dexamethasone . Phorbol ester also upregulated immunoreactive P35354 . Constitutive P35354 in these tissues has important implications for side effects of P35354 -selective inhibitors . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Combination of virtual screening and high throughput gene profiling for identification of novel liver X receptor modulators . We conducted virtual docking studies using GLIDE with modified LXRbeta ligand-binding domain ( LBD ) on internal compound collection followed by the gene profiling with ArrayPlate mRNA assay . A total of 69 compounds were found to upregulate LXRalpha and certain LXR regulated genes from 1308 compounds selected by virtual screen ( hit rate : 5.3 % ) . Compound 4 was shown to significantly induce the expression of LXR target genes such as O95477 , P45844 , P02649 , O00767 -1 , and SREBP-1c in THP-1 differentiated macrophages . In vitro binding assay confirmed that 4 binds to both LXRalpha and LXRbeta directly and recruits coactivator peptide Q15788 . It functions as a full LXR agonist in stimulating cholesterol efflux in THP-1 differentiated macrophages and induces lipogenesis in HepG2 cells . This study demonstrates that the combination of virtual screen and high throughput gene profiling is an efficient approach for rapid identification of novel LXR modulators . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Organic anion transporting polypeptide-C mediates arsenic uptake in P29320 -293 cells . Arsenic is an established human carcinogen . The role of aquaglyroporins ( AQPs ) in arsenic disposition was recently identified . In order to examine whether organic anion transporting polypeptide-C ( Q9Y6L6 ) also plays a role in arsenic transport , Q9Y6L6 cDNA was transfected into cells of a human embryonic kidney cell line ( P29320 -293 ) . Transfection increased uptake of the model Q9Y6L6 substrate , estradiol-17beta-D-glucuronide , by 10-fold . In addition , we measured uptake and cytotoxicity of arsenate , arsenite , monomethylarsonate(MMA(V)) , and dimethylarsinate ( P28067 (V) ) . Transfection of Q9Y6L6 increased uptake and cytotoxicity of arsenate and arsenite , but not of MMA(V) or P28067 (V) . DB01045 and taurocholic acid ( a substrate of Q9Y6L6 ) reversed the increased toxicity of arsenate and arsenite seen in Q9Y6L6 -transfected cells . The increase in uptake of inorganic arsenic was not as great as that of estradiol-17beta-D-glucuronide . Our results suggest that Q9Y6L6 can transport inorganic arsenic in a ( DB00143 ) -dependent manner . However , this may not be the major pathway for arsenic transport . Selective serotonin reuptake inhibitors attenuate the antigen presentation from dendritic cells to effector T lymphocytes . DB00472 , one of the selective serotonin reuptake inhibitors ( SSRIs ) , has been found to possess immune modulation effects , in addition to its antidepressant effects . However , it remains unclear whether SSRIs can suppress the antigen-presenting function of dendritic cells ( DCs ) . Therefore , DB00472 was applied to a co-culture of Aggregatibacter actinomycetemcomitans ( Aa ) -reactive T cells ( ×Aa-T ) isolated from Aa-immunized mice and DCs . This resulted in the suppressed proliferation of ×Aa-T stimulated with Aa-antigen presentation by DCs . Specifically , DB00472 increased the extracellular 5-hydroxytryptamine ( 5-HT ) in the ×Aa-T/DC co-culture , whereas exogenously applied 5-HT promoted T-cell proliferation in the ×Aa-T/DC co-culture , indicating that DB00472 -mediated suppression of ×Aa-T/DC responses can not be attributed to extracellular 5-HT . Instead , DB00472 remarkably suppressed the expression of costimulatory molecule Q9Y6W8 -L on DCs . DB00472 also promoted a greater proportion of P42081 (Low) immature DCs than P42081 (High) mature DCs , while maintaining the expression levels of P33681 , MHC-class-II and Q9NZQ7 . These results suggested that DB00472 suppressed the ability of DCs to present bacterial antigens to T cells , and the resulting T-cell proliferation , in a P31645 /5-HT-independent manner and that diminished expression of Q9Y6W8 -L on DCs and increase of P42081 (Low) immature DCs caused by DB00472 might be partially associated with DB00472 -mediated suppression of DC/T-cell responses . Expression patterns and role of prostaglandin-endoperoxide synthases , prostaglandin E synthases , prostacyclin synthase , prostacyclin receptor , peroxisome proliferator-activated receptor delta and retinoid x receptor alpha in rat endometrium during artificially-induced decidualization . To determine if changes in endometrial expression of the enzymes and receptors involved in prostaglandin ( PG ) synthesis and action might provide insights into the PGs involved in the initiation of decidualization , ovariectomized steroid-treated rats at the equivalent of day 5 of pseudopregnancy were given a deciduogenic stimulus and killed at various times up to 32 h thereafter . The expression of PG-endoperoxide synthases ( P23219 and P35354 ) , microsomal PGE synthases ( O14684 and Q9H7Z7 ) , cytosolic PGE synthase ( Q15185 ) , prostacyclin synthase ( Q16647 ) , prostacyclin receptor , peroxisome proliferator-activated receptor delta ( Q03181 ) and retinoid x receptor alpha ( P19793 ) in endometrium was assessed by semiquantitative RT-PCR , western blot analyses and immunohistochemistry . In addition , to determine which PG is involved in mediating decidualization , we compared the ability of PGE(2) , stable analogues of P06744 (2) , L165041 ( an agonist of Q03181 ) , and docasahexanoic acid ( an agonist of P19793 ) to increase endometrial vascular permeability ( EVP , an early event in decidualization ) , and decidualization when infused into the uterine horns of rats sensitized for the decidual cell reaction ( DCR ) . EVP was assessed by uterine concentrations of Evans blue 10 h after initiation of infusions . DCR was assessed by the uterine mass 5 days after the initiation of the infusions . Because enzymes associated with the synthesis of PGE(2) , including P35354 , are up-regulated in response to a deciduogenic stimulus and because PGE(2) was more effective than the P06744 (2) analogues and Q03181 and P19793 agonists in increasing EVP and inducing decidualization , we suggest that PGE(2) is most likely the PG involved in the initiation of decidualization in the rat . Toxicogenetics of antiretroviral therapy : genetic factors that contribute to metabolic complications . Metabolic complications of antiretroviral therapy ( O00253 ) have emerged as a major concern for long-term , successful management of HIV infection . Variability in the response to O00253 between individuals has been increasingly linked to the genetic background of patients , as regards efficacy and susceptibility to adverse reactions ( toxicogenetics ) . This review summarizes the biological and methodological background for the genetic prediction of metabolic toxicity of O00253 . Recent studies are discussed which suggest that single-nucleotide polymorphisms ( SNPs ) in several genes involved in lipid metabolism and lipid transport in the general population ( O95477 , Q6Q788 , P02656 , P02649 , P11597 ) might modulate plasma triglyceride and high-density lipoprotein cholesterol levels in HIV-infected patients . At present , genetic prediction of lipodystrophy is not possible . Lipodystrophy has been linked to an accumulation of mtDNA mutations , a finding causally associated with ageing phenotypes in animal models . No mutations in P02545 , a gene linked to rare , inherited forms of lipodystrophy , have been identified in small studies of patients with lipodystrophy , and a possible link to a P01375 promoter SNP remains to be confirmed . With the rapidly decreasing cost of genetic testing , the main issues that need to be addressed prior to introduction of toxicogenetic prediction in HIV clinical practice include reproducibly high predictive values of SNP associations with clinically relevant and well defined metabolic outcomes , studies that evaluate the contribution of SNPs in the context of multi-SNP and haplotype analysis , and the validation of genetic markers in independent , large patient cohorts . Comprehensive , whole genome approaches are increasingly being used . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Regulation of P61073 gene expression in breast cancer cells under diverse stress conditions . Chronic inflammation is a critical component in breast cancer progression . Pro-inflammatory mediators along with growth/survival factors within the tumor microenvironment potentiate the expression of pro-inflammatory cytokines ( IL-1 , P05231 , P01375 -α ) , chemotactic cytokines and their receptors ( P61073 , P48061 , P10145 ) and angiogenic factors ( P15692 ) that often overcome the effect of anti-inflammatory molecules ( P05112 , P22301 ) thus evading the host 's antitumor immunity . Detailed knowledge , therefore , of the regulatory mechanisms determining cytokine levels is essential to understand the pathogenesis of breast cancer . HIF-1α and NF-κB transcription factors are important players for the establishment of a pro-inflammatory and potentially oncogenic environment . HIF-1α is the key mediator of the cellular response to oxygen deprivation and induces the expression of genes involved in survival and angiogenesis within solid hypoxic tumors . The expression of these genes is often modulated by the p53 tumor suppressor protein that induces apoptosis or cell cycle arrest in neoplastic cells . Functional crosstalk between HIF-1α and p53 pathways mediated by modulators shared between the two transcription factors such as Q15788 and SIRT-1 differentially regulate the expression of distinct subsets of their target genes under variable stress conditions . In an attempt to shed light on the complex regulatory mechanisms involved in cancer-related inflammation , we investigated the role of the two common p53 and HIF-1α co-regulators Q15788 and SIRT-1 , in the expression of the highly potent metastatic chemokine receptor P61073 . Both Q15788 and SIRT-1 overexpression in DSFX-treated MCF-7 cells reduced P61073 cellular levels implying that both co-regulators are crucial factors in the determination of the metastatic potential of breast cancer cells . Comparison of the efficacy of peripheral blood stem cell mobilization using G- P04141 alone from healthy donors and patients with hematologic malignancies . Peripheral blood stem cell ( PBSC ) collection using granulocyte colony-stimulating factor ( DB00099 ) alone is superior to the combination of chemotherapy and G- P04141 in terms of low morbidity , short duration of mobilization and low cost . We retrospectively compared the results of PBSC collection using G- P04141 alone in 11 patients with malignant lymphoma ( ML ) , 23 patients with plasma cell neoplasms ( Q15149 ) and 48 healthy donors . The geometric mean number of P28906 (+) cells/kg obtained on the first day of collection was 0.99 × 10(6)/kg in ML patients , 2.26 × 10(6)/kg in Q15149 patients , and 3.36 × 10(6)/kg in healthy donors . The probability of collecting at least 1 × 10(6)/kg P28906 (+) cells/kg during a single course of apheresis was 90.9 % in ML patients , 95.7 % in Q15149 patients , and 100 % in healthy donors . In a multiple regression analysis of the P28906 (+) cell yields on the first day of apheresis , we identified disease , the baseline white blood cell count ( WBC ) , platelet count , and lactate dehydrogenase as independent significant variables . Particularly , disease was strongly associated with the P28906 (+) cell yield , probably due to the difference in the number of previous chemotherapy cycles . In conclusion , the minimal dose of P28906 (+) cells for autologous transplantation was collected in almost all patients with hematological malignancies . However , patients who have received repeated cycles of chemotherapy , such as patients with ML , and those who have low WBC counts and/or platelet counts may be at higher risk for poor mobilization . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . Rubella vaccine-induced cellular immunity : evidence of associations with polymorphisms in the Toll-like , vitamin A and D receptors , and innate immune response genes . Toll-like , vitamin A and D receptors and other innate proteins participate in various immune functions . We determined whether innate gene-sequence variations are associated with rubella vaccine-induced cytokine immune responses . We genotyped 714 healthy children ( 11-19 years of age ) after two doses of rubella-containing vaccine for 148 candidate SNP markers . Rubella virus-induced cytokines were measured by ELISA . Twenty-two significant associations ( range of P values 0.002-0.048 ) were found between SNPs in the vitamin A receptor family ( P10276 , P10826 , Q02880 and P13631 ) , vitamin D receptor and downstream mediator of vitamin D signaling ( P19793 ) genes and rubella virus-specific ( P01579 , P60568 , P22301 , P01375 , and GM- P04141 ) cytokine immune responses . A O15455 gene promoter region SNP ( rs5743305 , -8441A > T ) was associated with rubella-specific GM- P04141 secretion . Importantly , SNPs in the Q9C035 gene coding regions , rs3740996 ( His43Tyr ) and rs10838525 ( Gln136Arg ) , were associated with an allele dose-related secretion of rubella virus-specific P01375 and P60568 /GM- P04141 , respectively , and have been previously shown to have functional consequences regarding the antiviral activity and susceptibility to HIV-1 infection . We identified associations between individual SNPs and haplotypes in , or involving , the O95786 ( O95786 ) gene and rubella-specific P01375 secretion . This is the first paper to present evidence that polymorphisms in the TLR , vitamin A , vitamin D receptor , and innate immunity genes can influence adaptive cytokine responses to rubella vaccination . Genetic determinants of serum lipid levels in Chinese subjects : a population-based study in Shanghai , China . We examined the associations between 21 single nucleotide polymorphisms ( SNPs ) of eight lipid metabolism genes and lipid levels in a Chinese population . This study was conducted as part of a population-based study in China with 799 randomly selected healthy residents who provided fasting blood and an in-person interview . Associations between variants and mean lipid levels were examined using a test of trend and least squares mean test in a general linear model . Four SNPs were associated with lipid levels : P01130 rs1003723 was associated with total cholesterol ( P-trend = 0.002 ) and LDL ( P-trend = 0.01 ) , P01130 rs6413503 was associated with total cholesterol ( P-trend = 0.05 ) , P04114 rs1367117 was associated with apoB ( P-trend = 0.02 ) , and O95342 rs49550 was associated with total cholesterol ( P-trend = 0.01 ) , triglycerides ( P-trend = 0.01 ) , and apoA ( P-trend = 0.01 ) . We found statistically significant effects on lipid levels for P01130 rs6413503 among those with high dairy intake , P06858 rs263 among those with high allium vegetable intake , and P02649 rs440446 among those with high red meat intake . We identified new associations between SNPs and lipid levels in Chinese previously found in Caucasians . These findings provide insight into the role of lipid metabolism genes , as well as the mechanisms by which these genes may be linked with disease . Use of RNA interference to elucidate the effect of P04198 on cell cycle in neuroblastoma . BACKGROUND : P04198 amplification marks poor prognosis in neuroblastoma ( NB ) tumors . In evaluating the mechanisms by which retinoic acid ( RA ) or nerve growth factor ( P01138 ) decrease cell number in P04198 amplified NB cells , we have identified a number of proteins whose expression either decreases ( E2F , P06493 , Q00534 , cyclin dependent kinase activity ) or increases ( p27 ) in association with a decrease in P04198 expression . However , it was still unclear which were P04198 dependent effects or not . PROCEDURE : This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in P04198 . We silenced P04198 expression using siRNA targeted to the coding region of P04198 . Then , by using siRNA transient transfections , we analyzed the change of cell cycle related genes and cell cycle in P04198 amplified NB cell lines . RESULTS : We demonstrate that expression of P04198 can be suppressed by almost 60 % in P04198 amplified NB cell using siRNAs targeted to P04198 . Functionally , the decrease in P04198 leads to a decrease in cells in the S-phase of the cell cycle . Decreases in P04198 are associated with decreases in Q01094 -2 and Q02363 along with increases in p27 protein levels by post-transcriptional modification . Moreover , we find that a decrease in P04198 is accompanied by a decrease in cdk6 mRNA and protein expression . CONCLUSIONS : These results show that E2F and Q02363 expression is associated with P04198 regulation and that cdk6 is a possible new transcriptional target of P04198 . Quantification of rifampicin in human plasma and cerebrospinal fluid by a highly sensitive and rapid liquid chromatographic-tandem mass spectrometric method . A highly sensitive and rapid liquid chromatography tandem mass spectrometry ( LC-MS/MS ) method has been developed to measure the levels of the antitubercular drug rifampicin ( Q9HBH0 ) in human plasma and cerebrospinal fluid ( P04141 ) . The analyte and internal standard ( IS ) were isolated from plasma and P04141 by a simple organic solvent based precipitation of proteins followed by centrifugation . Detection was carried out by electrospray positive ionization mass spectrometry in the multiple-reaction monitoring ( MRM ) mode . The assay was linear in the concentration range 25-6400 ng/mL with intra- and inter-day precision of < 7 % and < 8 % , respectively . The validated method was applied to the study of Q9HBH0 pharmacokinetics in human P04141 and plasma over 25 h period after a 10 mg/kg oral dose . P35354 inhibitor treatment enhances photodynamic therapy-mediated tumor response . Photodynamic therapy ( PDT ) continues to be used in the treatment of solid tumors . Clinical results are promising , but the therapy has not been optimized , and tumor recurrences can occur . Recently , it has been shown that inhibitors of cyclooxygenase ( P36551 ) -2 can be effective in combination with conventional chemotherapy and radiation therapy . In the current study , we examined the parameters of PDT-mediated activation of P35354 expression . We also examined the tumoricidal effectiveness of combining PDT with the selective P35354 inhibitor NS-398 . PDT induced the transcriptional activation of P35354 . Prolonged expression of P35354 protein was observed in PDT-treated mouse sarcoma and carcinoma cell lines , whereas P23219 was not inducible by PDT . Prostaglandin ( PG ) E2 synthesis was also increased in PDT-treated cells , and DB00917 levels were attenuated in cells coincubated with NS-398 , indicating that PDT induced the expression of biologically active P35354 . Both porphyrin- and chlorin-based photosensitizers were able to elicit PDT-mediated P35354 expression . P35354 was also elevated in radiation-induced fibrosarcoma ( Q9HBH0 ) tumors after treatment with PDT . We also observed that systemic administration of NS-398 decreased PDT induction of both DB00917 and vascular endothelial growth factor in treated Q9HBH0 tumors . Additionally , we demonstrated that NS-398 enhanced PDT responsiveness in Q9HBH0 tumors without increasing toxicity to normal tissue . These results provide strong evidence that combination procedures involving selective P35354 inhibitors may improve the therapeutic effectiveness of PDT . P05231 and MYC collaborate in plasma cell tumor formation in mice . P05231 ( P05231 ) plays a critical role in the natural history of human plasma cell neoplasms ( PCNs ) , such as plasma cell myeloma and plasmacytoma ( PCT ) . P05231 is also at the center of neoplastic plasma cell transformation in BALB/c ( C ) mice carrying a transgene , H2-L(d)- P05231 , that encodes human P05231 under control of the major histocompatibility complex H2-L(d) promoter : strain C.H2-L(d)- P05231 . These mice are prone to PCT , but tumor development is incomplete with long latencies ( approximately 40 % PCT at 12 months of age ) . To generate a more robust mouse model of P05231 -dependent Q15149 , we intercrossed strain C.H2-L(d)- P05231 with strains C.iMyc(Emu) or C.iMyc(Calpha) , 2 interrelated gene-insertion models of the chromosomal T(12;15) translocation causing deregulated expression of Myc in mouse PCT . Deregulation of MYC is also a prominent feature of human Q15149 . We found that double-transgenic C.H2-L(d)- P05231 /iMyc(Emu) and C.H2-L(d)- P05231 /iMyc(Calpha) mice develop PCT with full penetrance ( 100 % tumor incidence ) and short latencies ( 3-6 months ) . The mouse tumors mimic molecular hallmarks of their human tumor counterparts , including elevated P05231 /Stat3/Bcl-X(L) signaling . The newly developed mouse strains may provide a good preclinical research tool for the design and testing of new approaches to target P05231 in treatment and prevention of human PCNs .

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DB00065 -Induced Hypothyroidism : A Novel Case and Postulations concerning the Mechanism . We report a patient with cutaneous sarcoidosis who developed hypothyroidism following 17 months of infliximab therapy . To our knowledge , this is the first reported case of hypothyroidism following infliximab administration . While it is possible that the patient 's hypothyroidism was unrelated to the use of infliximab , the time course and lack of alternative explanations make such an association plausible . We postulate that hypothyroidism in this patient may have been related to the development of autoantibodies to infliximab that triggered the development of an autoimmune thyroiditis . Regardless of the mechanism , we would encourage clinicians to keep the potential mechanisms of P01375 -α in mind when treating patients with P01375 -α antagonist medications . DB00065 and nephrotic syndrome . DB00065 is a chimeric tumor necrosis factor-alpha ( P01375 ) monoclonal antibody , which has been used extensively in patients with rheumatoid arthritis and inflammatory bowel disease . It also appears to be effective in other conditions such as psoriasis and ankylosing spondylitis . The major side effect of infliximab is infection . Renal complications are uncommon and not well recognized . This report describes a probable case of infliximab-induced membranous nephropathy . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . DB00065 dependency in children with Crohn 's disease . BACKGROUND : Recently , infliximab dependency has been described . AIM : To assess frequency of ID in 82 consecutive Crohn 's disease children treated with infliximab 2000-2006 and to describe clinical and genetic predictors of long-term infliximab response . METHODS : A phenotype model of infliximab dependency was used to assess treatment response : ' immediate outcome ' ( 30 days after infliximab start ) -- complete/partial/no response . ' Long-term outcome ' : ( i ) prolonged response : maintenance of complete/partial response ; ( ii ) infliximab dependency : relapse < or = 90 days after intended infliximab cessation requiring repeated infusions to regain complete/partial response or need of infliximab > 12 months to sustain response . Polymorphisms P01375 -308 A > G , P01375 -857 C > T , Casp9 93 C > T , P48023 -844 C > T , P01374 252 C > T and Q9HC29 ( R702W , G908R , 1007fs ) were analysed . RESULTS : Ninety-four per cent of children obtained complete/partial response . In long-term outcome , 22 % maintained prolonged response , 12 % had no response , while 66 % became infliximab dependent . Perianal disease and no previous surgery were associated with infliximab dependency ( OR 5.34 , 95 % CI : 1.24-22.55 ; OR 6.7 , 95 % CI : 1.67-26.61 ) . No association was found with studied polymorphisms . The cumulative probability of surgery 50 months after starting infliximab was 10 % in infliximab dependency , 30 % in prolonged responders and 70 % in nonresponders ( P = 0.0002 ) . CONCLUSIONS : Sixty-six per cent of children became infliximab dependent . Perianal disease and no surgery prior to infliximab were associated with infliximab dependency phenotype . Anti-tumor necrosis factor-alpha therapy in the ordinary clinical setting : Three-year effectiveness in patients with rheumatoid arthritis . OBJECTIVES : The aim was to estimate the proportion of patients with rheumatoid arthritis ( RA ) achieving low disease activity by anti-tumor necrosis factor-alpha ( P01375 ) therapy in an ordinary clinical setting . METHODS : Thirty-three patients with active RA despite methotrexate treatment were included in an open phase IV study of infliximab in combination with methotrexate . The mean age was 53years ( range 21-71 ) and mean disease duration 10.7years ( 1-32 ) . Treatment was changed in cases of insufficient response or intolerable adverse events . Response status to infliximab was assessed according to the American College of Rheumatology ( P10323 20 ) . Disease activity score ( DAS28 ) was assessed at baseline and at weeks 26 , 54 , 80 , 106 and 158 . Low disease activity is defined as DAS28 < or=3.2 . RESULTS : ACR20 response to infliximab was recorded in 73 % and 33 % of the patients at weeks 26 and 158 , respectively . DB00065 was discontinued in 16 patients , 15 of whom started other anti- P01375 therapy . The baseline DAS28-score of 6.3 ( 95 % CI 5.9-6.6 ) was significantly reduced after 26weeks to 4.1 ( 95 % CI 3.6-4.7 ) and was later nearly stable . Low DAS28 was achieved by 36 % of the patients at week 26 , and by 15-24 % at later assessments . CONCLUSIONS : RA disease activity was significantly reduced , but DAS28 < or=3.2 was recorded in only 1/4 of the assessments . Increasing this proportion should be subject to continuous quality improvement efforts . Phenotype and functional changes of Vgamma9/Vdelta2 T lymphocytes in Behçet 's disease and the effect of infliximab on Vgamma9/Vdelta2 T cell expansion , activation and cytotoxicity . INTRODUCTION : DB00065 is a chimeric monoclonal antibody against tumor necrosis factor alpha ( P01375 ) that has been introduced recently for Behçet 's disease ( BD ) patients who were resistant to standard treatment . The aim of this study was to analyse the functional changes of Vgamma9/Vdelta2 T lymphocytes in both active and inactive disease and the effect of infliximab on Vgamma9/Vdelta2 T cell expansion , activation and cytotoxicity . METHODS : We investigated 1 ) cell expansion , 2 ) expression of TNFRII receptor , 3 ) perforin and gamma interferon ( IFN ) content , 4 ) release of granzyme A ( GrA ) and 5 ) phenotype changes , in vitro and in vivo , in Vgamma9/Vdelta2 T lymphocytes by means of fluorescence-activated cell sorter analysis of lymphocyte cultures from patients with active and inactive BD and healthy subjects . RESULTS : Cell expansion , expression of TNFRII , perforin and gamma IFN content and release of granzyme A were significantly higher in active patients . In vitro and ex vivo treatment with infliximab resulted in a significant reduction of all parameters together with changes in the phenotype of Vgamma9/Vdelta2 T cells . CONCLUSIONS : All together these data indicate that infliximab is capable of interfering with Vgamma9/Vdelta2 T cell function in BD and although cell culture models can not reliably predict all potential effects of the drug in vivo , our results present the possibility that this drug may find use in a range of immunological disorders , characterized by dysregulated cell-mediated immunity . Response to infliximab therapy in ulcerative colitis is associated with decreased monocyte activation , reduced P13500 expression and downregulation of P24821 C . BACKGROUND AND AIMS : The cellular mechanisms leading to infliximab therapy response in patients with ulcerative colitis ( UC ) are incompletely known . We therefore investigated early effects of infliximab therapy on monocytes and associated chemokines linked to clinical therapy response in UC patients . METHODS : Blood and biopsies were obtained from anti- P01375 therapy-naïve UC patients ( n = 43 ) before ( baseline ) and during induction therapy with infliximab . Therapy response was evaluated at Week 14 . Expression of monocyte activation markers and levels of chemokines in serum and biopsies were determined . Quantitative proteomic analysis was performed in cultured mucosal biopsies , and obtained data was validated in serum . RESULTS : In therapy responders , but not in non-responders , infliximab reduced blood monocyte expression of P08571 and P42081 , 2 weeks after therapy commenced , relative to baseline . Serum P13500 levels were decreased only among therapy responders at Week 2 and Week 14 , relative to baseline . These data corresponded with lower levels of P08571 , P42081 and P13500 in intestinal tissue in responders as compared with non-responders at Week 14 . Proteomic analysis of cultured biopsies showed that infliximab induced a reduction in P24821 C that predicted downregulation of P13500 . Therapy responders , but not non-responders , had decreased serum P24821 C levels at Week 2 and Week 14 , relative to baseline . CONCLUSIONS : DB00065 therapy response in UC patients is associated with reduced monocyte activation and serum levels of P13500 2 weeks after therapy commencement . In therapy responders , infliximab influenced P24821 C , which might be a regulator of P13500 expression and important for induction of the clinical therapy response . P01375 -alpha blockade for the treatment of acute GVHD . Despite posttransplantation immunosuppressive therapy , acute graft-versus-host disease ( GVHD ) remains a major cause of sickness and death . P01375 -alpha ( P01375 ) is implicated in the pathophysiology of GVHD at several steps in the process . DB00065 is a genetically constructed immunoglobulin P55008 ( IgG1 ) murine-human chimeric monoclonal antibody that binds the soluble subunit and the membrane-bound precursor of P01375 , blocking its interaction with receptors and causing lysis of cells that produce P01375 . In this study we retrospectively evaluated 134 patients who had steroid-refractory acute GVHD . Of these , 21 who received infliximab as a single agent were analyzed . The overall response rate was 67 % ( n = 14 ) , and 13 patients ( 62 % ) experienced complete response ( CR ) . Five patients ( 24 % ) did not respond , and 2 ( 10 % ) had progressive GVHD . None had a toxic reaction to infliximab . Ten patients ( 48 % ) had 18 fungal infections , including Aspergillus species in 7 and Candida species in 10 . Seventeen patients ( 81 % ) had bacterial infections , including 32 gram-positive and 8 gram-negative infections . Viral infections , primarily cytomegalovirus reactivation , occurred in 14 patients ( 67 % ) . The Kaplan-Meier estimate of overall survival was 38 % . In conclusion , infliximab was well tolerated and active for the treatment of steroid-resistant acute GVHD , particularly with gastrointestinal tract involvement . Survival after steroid-resistant acute GVHD continues to be problematic . The possibility of excessive fungal and other infections must be explored further . Behçet 's syndrome : a critical digest of the recent literature . A similar disease severity among men and women in Brasil , a high frequency of gastrointestinal involvement in China , Japan and USA , a low frequency of pathergy positivity in Japan and USA underline the ethnic variations reported in recent studies . Polymorphisms pertaining both to innate and adaptive immunity in genome wide association studies , clusters in phenotype , and new mechanisms for emerging therapeutic implications have been reported . A Th17 dominance seems to be likely with the exception of gastrointestinal involvement . DB00065 , interferon-alpha and cyclosporine-A may be showing their beneficial effects also by affecting the Th17 cells . The clinical course and outcome of isolated pulmonary artery thrombosis is similar to pulmonary artery aneurysms . Parenchymal lesions ( nodules , consolidations , cavities and ground glass lesions ) are common in patients with pulmonary involvement . Pericarditis is a frequent cardiac manifestation in France . Treatment of BS became more intensive than before . Immunosuppressives and corticosteroids seem to prevent relapses of venous thrombosis . Studies are needed to understand the role of anticoagulants . DB00034 appears to be effective at lower dosage , which brings the advantage of decreased cost and increased tolerability . Switching between anti- P01375 agents , when needed , is possible . Interleukin-1 and interleukin-6 are new promising targets . The risks of post-operative complications following pre-operative infliximab therapy for Crohn 's disease in patients undergoing abdominal surgery : a systematic review and meta-analysis . BACKGROUND : DB00065 is an anti- P01375 alpha blocker frequently utilized in the management of moderate to severe Crohn 's Disease . The immunosuppressive effects of infliximab may increase the risk for post-operative complications among Crohn 's Disease patients undergoing abdominal surgery . We conducted a systematic review and meta-analysis of studies comparing the rates of post-operative complications among Crohn 's disease patients treated with DB00065 therapy versus alternative therapies . METHODS : We used the Preferred Reporting Items for Systematic Reviews and Meta-Analyses ( PRISMA ) and searched 4 electronic databases along with major conference abstract databases from inception of database until November , 2012 . English-language articles and abstracts evaluating post-operative complications among Crohn 's disease patients were considered eligible . We applied meta-analysis with random effects model to calculate the overall odds ratio for total major complications as well as several secondary outcomes . RESULTS : Data were extracted from six studies including 1159 patients among whom 413 complications were identified . The most common complications were wound infections , anastomotic leak and sepsis . There was no significant difference in the major complication rate ( OR=1.59 [ 95 % CI : 0.89-2.86 ] ; p=0.15 ) , minor complication rate ( OR=1.80 [ CI : 0.87-3.71 ] ; p=0.11 ) , reoperation rate ( OR=1.33 [ CI : 0.55-3.20 ] ; p=0.52 ) or 30 day mortality rate ( OR=3.74 [ CI : 0.56-25.16 ] ; p=0.13 ) between the DB00065 and control groups . CONCLUSIONS : This meta analysis provides some evidence that infliximab may be safe to continue in the pre-operative period without increasing the risk of post-operative complications for Crohn 's disease patients undergoing abdominal surgery . [ Pharmacological profile of anti-human P01375 alpha monoclonal antibody , infliximab ( Remicade ) ] . P01375 alpha ( tumor necrosis factor-alpha ) plays an important role in the pathogenesis of inflammatory diseases including Crohn 's disease and rheumatoid arthritis . DB00065 ( Remicade ) is a chimeric monoclonal antibody that recognizes human P01375 alpha . Clinical trials trials have been persuasive that infliximab is effective in both Crohn 's disease and rheumatoid arthritis . DB00065 is an important treatment option in patients with active Crohn 's disease who have not responded to conventional therapy and in patients with Crohn 's disease who have fistulae . Moreover , infliximab plus methotrexate is effective in patients with active rheumatoid arthritis who have not responded adequately to traditional disease-modifying anti-rheumatic drugs , in terms of reducing symptoms and signs , inhibiting the progression of structural damage and improving physical function . Cutaneous adverse reaction to infliximab : report of psoriasis developing in 3 patients . DB00065 is a chimeric immunoglobulin G1kappa monoclonal antibody against tumor necrosis factor alpha ( P01375 ) , a proinflammatory cytokine that participates in both normal immune function and the pathogenesis of many autoimmune disorders . Treatment with infliximab reduces the biologic activities of P01375 and thus is indicated in the treatment of rheumatoid arthritis , Crohn disease , ankylosing spondylitis , psoriatic arthritis , plaque psoriasis , and ulcerative colitis . To our knowledge , there have been 13 case reports of new-onset psoriasis , psoriasiform dermatitis , and palmoplantar pustular psoriasis that developed during treatment with infliximab . We report 3 additional cases of biopsy-proven new-onset psoriasis that developed while the patients underwent treatment with infliximab for inflammatory bowel disease . Although the mechanism for the development of psoriasis in these patients is unclear , several possible explanations are proposed . With increasing use of infliximab and other P01375 inhibitors in clinical practice , more cases of similar reactions to these drugs probably will be reported and are necessary to determine the importance of this eruption . Optimizing anti-tumor necrosis factor strategies in inflammatory bowel disease . The introduction of infliximab , a mouse/human chimeric monoclonal antibody to tumor necrosis factor ( P01375 ) , is an important advance in the treatment of Crohn 's disease . DB00065 is effective for induction and maintenance of remission in patients with inflammatory luminal and fistulizing disease . The development of human antichimeric antibodies ( HACAs ) has led to infusion reactions and loss of efficacy in patients treated with infliximab . Strategies to reduce the frequency of HACA formation include induction of immunologic tolerance with a three-dose regimen at 0 , 2 , and 6 weeks followed by systematic maintenance dosing every 8 weeks ; concomitant immunosuppressive therapy with azathioprine , 6-mercaptopurine , or methotrexate ; and premedication with intravenous corticosteroids . Humanized or fully human anti- P01375 biotechnologic agents , including CDP571 , DB08904 , etanercept , adalimumab , and onercept , are theoretically less immunogenic than the chimeric antibody infliximab . DB00005 is not effective for Crohn 's disease . CDP571 is not effective in unselected patients with active Crohn 's disease , but it may be effective in patients with elevated P02741 . The efficacy of DB08904 , adalimumab , and onercept is under investigation . The different mechanisms of action of these anti- P01375 agents may account for their variable efficacy . Their benefits , however , must be considered in the context of their risks , including infusion reaction ; delayed hypersensitivity-like reaction ; new onset of autoimmunity , with rare cases of drug-induced lupus and new-onset demyelination ; and the potential for rare but serious infections . Development of new-onset psoriasis in a patient receiving infliximab for treatment of rheumatoid arthritis . DB00065 is a chimeric monoclonal antibody that selectively blocks tumor necrosis factor-alpha ( P01375 ) . It is indicated for the treatment of numerous inflammatory diseases , including rheumatoid arthritis , Crohn disease , ulcerative colitis , ankylosing spondyilitis , and psoriatic arthritis . DB00065 has also been shown to be a well tolerated and highly effective treatment for psoriatic skin lesions . We report an interesting case of unexpected , new-onset psoriasis in a patient on infliximab for rheumatoid arthritis . Refractory Wegener 's granulomatosis responds to tumor necrosis factor blockade . Wegener 's granulomatosis is a systemic necrotising vasculitis of small vessels that leads to severe impairment of affected organ systems . Conventional treatment is based on immunosuppression with a combination of steroids , cyclophosphamide , azathioprine or methotrexate over a prolonged time course . Early recurrence or disease refractory to therapy often results in a fatal outcome . As in other inflammatory disorders , tumor necrosis factor ( P01375 ) plays an early and crucial role in progression of disease activity . We report on a patient with severe orbital Wegener 's granulomatosis who developed acute renal failure despite intense conventional immunosuppression with cyclophosphamide and steroids . To stop vasculitic activity , by disrupting the autoimmune inflammatory cascade , a P01375 -blocking antibody ( DB00065 ) was administered six times in a six-month period at 3 mg/kg body weight . Conventional immunosuppressive therapy with steroids and cyclophosphamide was continued , the latter being changed to azathioprine after three months . The first infusion of P01375 antibody induced improvement of renal function , which continued throughout the course of therapy . The modification of diet in renal disease-glomerular filtration rate ( MDRD- Q92565 ) increased from 15.3 ml/min/1.73 m2 before the start of P01375 -blockade to 55.5 ml/min/1.73 m2 after six months of therapy . Serum creatinine levels , proteinuria and cANCA titer decreased concomitantly . Clinical remission of Wegener 's granulomatosis was induced without any major adverse events . A slight flare of orbital inflammation was successfully treated with an increased dose of azathioprine . Thus , in this case of refractory Wegener 's granulomatosis P01375 -blockade by monoclonal chimeric P01375 -antibody ( DB00065 ) served as an effective tool to rescue kidney function and induce clinical remission . Can we modulate the clinical course of inflammatory bowel diseases by our current treatment strategies ? Ulcerative colitis and Crohn 's disease are chronic disabling lifelong diseases which may be disturbed by severe flares and anatomical complications requiring surgery . Until the very last years there was no clear indication that treatment was able to modify the long-term natural history of the disease . In particular , there are no data demonstrating a clear improvement through the period 1950-2003 in disease activity , occurrence of complications and need for surgery , in spite of an increased use of immunosuppressants since the 1990s . However , in inflammatory bowel disease , both thiopurines and methotrexate are very efficient in about one half of the patients , and in responders , may heal the mucosa and decrease the need for surgery . The early use of immunosuppressants in selected patients may have an impact on occurrence of severe flares and complications , and need for surgery . Moreover , anti- P01375 now used for 10 years in Crohn 's disease and for 5 years in ulcerative colitis demonstrated in two thirds of the patients a remarkable anatomic effect , healing the mucosa , closing fistulae and preventing strictures . DB00065 does prevent endoscopic recurrence following ileal resection for Crohn 's disease . Actually , because irreversible anatomical damage may develop during the first years of disease , there is a need to classify early in the course of the disease patients who will benefit from anti- P01375 and classical immunosuppressants , respectively . There is the need in the next few years to better define these subgroups and to compare different strategies within each group through randomized interventional studies . Biological therapies in Crohn 's disease : are they cost-effective ? A critical appraisal of model-based analyses . In refractory Crohn 's disease , anti- P01375 and anti-α 4 integrin agents are used for ameliorating disease activity but impose high costs to health-care systems . The authors systematically reviewed cost-effectiveness analyses based on decision models : most of the studies were judged to have a good quality , but a large portion assessed health and costs in a short time horizon , usually disregarding fistulizing disease and not considering safety . DB00065 induction followed by on-demand retreatment consistently proved to have a good cost per quality-adjusted life year , while maintenance treatment never satisfied commonly accepted cost-utility thresholds . Challenges in cost-effectiveness analysis include the lack of a standard model structure , a large variability in the costs of surgery and poor data on indirect costs . As clinical practice is moving to mucosal healing as a robust response marker , personalized schedules of anti- P01375 therapies might prove cost-effective even in the perspective of the health-care system in the near future . DB00065 counteracts tumor necrosis factor-α-enhanced induction of matrix metalloproteinases that degrade claudin and occludin in non-pigmented ciliary epithelium . DB00065 , a monoclonal antibody directed against human tumor necrosis factor-alpha ( P01375 -α ) , effectively treats anterior uveitis , which can accompany Behçet 's disease . Here , we investigated the underlying mechanism of this action . We examined human , non-pigmented ciliary epithelial cells ( HNPCECs ) , which make up the blood-aqueous barrier ( BAB ) in the uvea . We measured the expression levels of matrix metalloproteinases ( MMPs ) and tissue inhibitors of MMPs in the presence or absence of P01375 -α using quantitative , real-time polymerase chain reaction and enzyme-linked immunosorbent assays . The expression of P03956 , P08254 , and P14780 increased in the presence of P01375 -α , and the addition of infliximab reversed the increase . The P01375 -α effects were more attenuated when infliximab was added before than when it was added after P01375 -α exposure . Gelatin zymography demonstrated that the protease activity of these MMPs was also increased in the presence of P01375 -α and attenuated with infliximab . Immunostaining showed that P03956 , P08254 , and P14780 degraded claudin-1 and occludin in HNPCECs and in non-pigmented ciliary epithelial cells of the swine ciliary body . In a monolayer of HNPCECs , we found that permeability was significantly increased with MMP treatment . Thus , P01375 -α increased levels of MMPs in cells that form the BAB , and MMPs degraded components of the tight junctions in the BAB , which increased permeability through the cellular barrier . Furthermore , infliximab effectively attenuated the P01375 -α-induced increases in MMP expression in cells that make up the BAB . These findings might suggest a basis for the clinical prevention of anterior uveitis . DB00065 in active early rheumatoid arthritis . OBJECTIVE : To examine the impact of the combination of infliximab plus methotrexate ( MTX ) on the progression of structural damage in patients with early rheumatoid arthritis ( RA ) . METHODS : Subanalyses were carried out on data for patients with early RA in the Anti- P01375 Therapy in RA with Concomitant Therapy ( ATTRACT ) study , in which 428 patients with active RA despite MTX therapy received placebo with MTX ( MTX-only ) or infliximab 3 mg/kg or 10 mg/kg every ( q ) 4 or 8 weeks with MTX ( infliximab plus MTX ) for 102 weeks . Early RA was defined as disease duration of 3 years or less ; 82 of the 428 patients ( 19 % ) met this definition . Structural damage was assessed with the modified van der Heijde-Sharp score . The changes from baseline to week 102 in total modified van der Heijde-Sharp score were compared between the infliximab plus MTX groups and the MTX-only group . RESULTS : The erosion and joint space narrowing scores from baseline to week 102 in the cohort of patients with early RA decreased significantly in each infliximab dose regimen compared with the MTX-only regimen . Consistent benefit was seen in the joints of both hands and feet . CONCLUSIONS : DB00065 combined with MTX inhibited the progression of structural damage in patients with early RA during the 2 year period of treatment . Early intervention with infliximab in patients with active RA despite MTX therapy may provide long term benefits by preventing radiographic progression and preserving joint integrity . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . DB00065 restores glucose homeostasis in an animal model of diet-induced obesity and diabetes . P01375 plays an important role in obesity-linked insulin resistance and diabetes mellitus by activating at least two serine kinases capable of promoting negative regulation of key elements of the insulin signaling pathway . Pharmacological inhibition of P01375 is currently in use for the treatment of rheumatoid and psoriatic arthritis , and some case reports have shown clinical improvement of diabetes in patients treated with the P01375 blocking monoclonal antibody infliximab . The objective of this study was to evaluate the effect of infliximab on glucose homeostasis and insulin signal transduction in an animal model of diabetes . Diabetes was induced in Swiss mice by a fat-rich diet . DB09341 and insulin homeostasis were evaluated by glucose and insulin tolerance tests and by the hyperinsulinemic-euglycemic clamp . Signal transduction was evaluated by immunoprecipitation and immunoblotting assays . Short-term treatment with infliximab rapidly reduced blood glucose and insulin levels and glucose and insulin areas under the curve during a glucose tolerance test . Furthermore , infliximab increased the glucose decay constant during an insulin tolerance test and promoted a significant increase in glucose infusion rate during a hyperinsulinemic-euglycemic clamp . In addition , the clinical outcomes were accompanied by improved insulin signal transduction in muscle , liver , and hypothalamus , as determined by the evaluation of insulin-induced insulin receptor , insulin receptor substrate-1 , and receptor substrate-2 tyrosine phosphorylation and Akt and forkhead box protein O1 serine phosphorylation . Thus , pharmacological inhibition of P01375 may be an attractive approach to treat severely insulin-resistant patients with type 2 diabetes mellitus . Paraplegic mice are leading to new advances in spinal cord injury research . STUDY DESIGN : Experimental laboratory investigations with paraplegic mouse models . OBJECTIVES : To review the most recent advances in the field of spinal cord injury research ; immune system response , regeneration , and functional recovery . SETTINGS : Laval University and Laval University Medical Center , Quebec , Canada . METHODS : Assessment of regenerative processes and locomotor function recovery induced by a variety of treatments and approaches in wild-type and genetically engineered mice with complete or incomplete lesions of the spinal cord . RESULTS : Recent studies have reported a number of significant observations providing additional insight into the role and mechanism of regeneration , immune system response , and functional recovery after spinal cord injury ( SCI ) using incomplete paraplegic mice with Nogo-A , Q9BZR6 , EphA4 , P14136 /vimentime , P15018 , or Fas gene knock-out . A novel antibody called P02778 was also recently found to increase tissue sparing and angiogenesis after SCI . In an attempt to explore the possibilities of reactivating spared neurons below the injury level , researchers have found that pharmacological activation of specific subtypes of serotonin receptors ( eg , P08908 /2A/7 ) can sustain the production of basic locomotor-like movements in complete paraplegic mice . CONCLUSION : The growing availability of genetically engineered and mutant mouse strains along with molecular biology tools has led scientists to increasingly use murine models in SCI research . These new tools and models may assist scientists in understanding further the complex pathological consequences of SCI . Redo Ileal pouch-anal anastomosis combined with anti- P01375 -α maintenance therapy for Crohn 's disease with pelvic fistula : report of two cases . Pouch failure has been reported to occur after ileal pouch-anal anastomosis for Crohn 's disease . We report two cases of patients with Crohn 's disease , who underwent redo ileal pouch-anal anastomosis ( redo-IPAA ) combined with anti- P01375 -α maintenance therapy , with good functional results . The first patient , a man with presumed ulcerative colitis , suffered pelvic fistula recurrence and anastomotic dehiscence . He underwent redo-IPAA , at which time longitudinal ulcers were found . DB00065 was started 4 days postoperatively and continued . The second patient , a woman treated for ulcerative colitis , underwent laparoscopic IPAA 8 years later . After the development of a pelvic fistula , twisted mesentery of the ileal pouch was found intraoperatively and Crohn 's disease was diagnosed . DB00051 therapy resulted in fistula closure . Redo-IPAA was performed to normalize the twisted mesentery of the ileal pouch . No complications have been observed in either patient , both of whom have experienced good functional results after closure of the covering stomas . P01375 blockade in the treatment of rheumatoid arthritis : infliximab versus etanercept . There is accumulating evidence that tumour necrosis factor ( P01375 ) plays a major role in the pathogenesis of rheumatoid arthritis ( RA ) . Recent biotechnological advances have allowed for the development of agents that directly target P01375 , a pro-inflammatory cytokine . In the last 2 years , the US FDA and the EU 's Commission of the European Communities have approved two biological agents for the treatment of refractory RA , etanercept and infliximab . DB00005 is a fusion protein , composed of the Fc portion of IgG1 and the extracellular domain of the P01375 receptor ( p75 ) . DB00065 is a chimeric monoclonal antibody ( mAb ) composed of murine variable and human constant regions . In placebo-controlled trials , both agents have proven to be effective and well-tolerated in RA patients . This review evaluates the available P01375 inhibitors , summarises pertinent clinical trials and underscores differences between the two agents in terms of molecular structure , efficacy , safety data , antigenicity and pharmacokinetics . Meta-analysis technique confirms the effectiveness of anti- P01375 in the management of active ulcerative colitis when administered in combination with corticosteroids . BACKGROUND : P01375 has an important role in the pathogenesis of ulcerative colitis ( UC ) . It therefore seems that infliximab , the antibody against P01375 , is beneficial in the treatment of UC . The aim was to determine whether infliximab induces clinical response and remission in patients with UC using the meta-analysis technique . MATERIAL/METHODS : The Pubmed and Embase databases were searched for studies investigating the efficacy of infliximab on UC . Data from 1966 to 2006 were collected . The keywords used to search were " ulcerative colitis " with " infliximab " , " anti tumor necrosis factor " , or " anti tumor necrosis factors " . The reference lists from the retrieved articles were also reviewed for additional applicable studies . RESULTS : The summary odds ratio ( OR ) for clinical remission in four studies was 3.24 with a 95 % CI of 1.6-6.57 and a significant OR . The summary OR for clinical response in three studies was 3.93 with a 95 % CI of 2.84-5.45 and a significant OR . CONCLUSIONS : DB00065 is effective in inducing response and remission in patients with ulcerative colitis when administered in combination with corticosteroids . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Citrullination of P01375 -α by peptidylarginine deiminases reduces its capacity to stimulate the production of inflammatory chemokines . Citrullination , a posttranslational modification ( PTM ) recently discovered on inflammatory chemokines such as interleukin-8 ( P10145 / P10145 ) and interferon-γ-inducible protein-10 ( P02778 / P02778 ) , seriously influences their biological activity . Citrullination or the deimination of arginine to citrulline is dependent on peptidylarginine deiminases ( PADs ) and has been linked to autoimmune diseases such as multiple sclerosis ( MS ) and rheumatoid arthritis ( RA ) . Chemokines are to date the first identified PAD substrates with receptor-mediated biological activity . We investigated whether cytokines that play a crucial role in RA , like interleukin-1β ( IL-1β ) and tumor necrosis factor-alpha ( P01375 -α ) , may be citrullinated by PAD and whether such a PTM influences the biological activity of these cytokines . IL-1β and P01375 -α were first incubated with PAD in vitro and the occurrence of citrullination was examined by Edman degradation and a recently developed detection method for citrullinated proteins . Both techniques confirmed that human P01375 -α , but not IL-1β , was citrullinated by PAD . Citrullination of P01375 -α reduced its potency to stimulate chemokine production in vitro on human primary fibroblasts . Concentrations of the inflammatory chemokines P10145 , P02778 and monocyte chemotactic protein-1 ( P13500 / P13500 ) were significantly lower in supernatants of fibroblasts induced with citrullinated P01375 -α compared to unmodified P01375 -α . However , upon citrullination P01375 -α retained its capacity to induce apoptosis/necrosis of mononuclear cells , its binding potency to DB00065 and its ability to recruit neutrophils to the peritoneal cavity of mice . DB00065 treatment for refractory kawasaki disease in korean children . BACKGROUND AND OBJECTIVES : This was a multicenter study to evaluate the usefulness of the tumor necrosis factor-alpha ( P01375 ) blocker infliximab for treatment of Korean pediatric patients with refractory Kawasaki disease ( KD ) . SUBJECTS AND METHODS : Data from 16 patients throughout Korea who were diagnosed with refractory KD and received infliximab were collected retrospectively . RESULTS : Complete response to therapy with cessation of fever occurred in 13 of 16 patients . P02741 ( CRP ) concentrations decreased following infliximab infusion in all 14 patients in whom it was measured before and after treatment . There were no infusion reactions or complications associated with infliximab except in 1 case with acute hepatitis occurring during treatment followed by calculous cholecystitis 4 months later . Fifteen patients had coronary artery ( CA ) abnormalities before infliximab therapy . Three had transient mild dilatation and 9 had CA aneurysms , with subsequent normalization in 4 patients , persistent mild dilatation in 3 , persistent aneurysm in 2 , and there were 3 cases ( 2 with CA aneurysm , 1 with mild CA dilatation ) without follow-up echocardiography . CONCLUSION : The results of this study suggest that infliximab may be useful in the treatment of refractory KD , and it appears that there is no significant further progression of CA lesions developing after infliximab treatment . Multicenter trials with larger numbers of patients and long-term follow-up are necessary to assess the clinical efficacy and safety of infliximab in refractory KD . [ Inflammatory bowel disease : from sulfasalazine to biologics ] . The arrival of biologics , in particular anti-Tumor Necrosis Facteur ( P01375 ) , at the end of the nineties , revolutionnized treatment of inflammatory bowel diseases . Concomitantly , immunosuppressants ( thiopurines , methotrexate ) are used more widely and earlier in the disease course . DB00065 and adalimumab are very effective in more than two-thirds of patients , including those with fistula . This efficacy is long lasting in one-third of patients . Main side-effects of anti- P01375 are opportunistic infections ( intracellular bacteria ) which should be prevented and diagnosed early . Anti- P01375 are safe in the long-term , however , there is a particular concern regarding the risk of hepatosplenic T cell lymphomas in young men receiving bitherapy with thiopurine and anti- P01375 . The old strategy of adapting the therapeutic response to severity of symptoms and disease activity has no impact on natural history of the disease and should be abandoned . Most authors now favour an aggressive therapeutic approach in selected patients , before they develop irreversible anatomic lesions . This new strategy may change natural history and will become safer with a better knowledge of side-effects of immunosuppressants and biologics and how to prevent them . Moreover development of new therapeutic agents may permit to avoid surgery in patients who do not respond to therapy . [ DB00065 treatment trial in a patient with neuro-Behçet 's disease unresponsive to other treatments ] . A 22-year-old man with a previous uveitis episode was admitted to our hospital because of persistent hiccup . On admission , he presented right-upper quadrantanopia , mydriasis and lack of the light reflex in the left eye , left-sided hemiplegia , and bilateral pathologic hyperreflexia . The MR fluid attenuated inversion recovery images showed left side dominant , high intensity lesions on the brainstem and the diencephalon . The HLA-B51 was positive . The P04141 P05231 was extremely elevated ( 998 pg/ml : reference value < = 6.0 pg/ml ) . Based on these , we concluded he had the neuro-Behçet 's disease and treated him by high dose intravenous corticosteroids . This treatment improved his symptoms and Q9BWK5 lesions , and decreased the P04141 P05231 levels initially . On 13th day after the first his discharge , however , dysarthria appeared and the P04141 P05231 levels elevated again . In addition to the high dose intravenous corticosteroids therapy for acute attack , 15 mg/week of methotrexate was started to prevent the recurrence . Even with this prevention , meningitis related to neuro-Behçet 's disease occurred within six weeks . We administered 5 mg/kg of infliximab intravenously at 0 , 2 , 6 , and 14 weeks . After the infliximab treatment , his symptoms improved and the P05231 levels decreased , and no recurrence has occurred . This case supports that infliximab , anti- P01375 agent , is a good candidate for neuro-Behçet 's disease treatment when it is resistant to conventional immunosuppressive agents such as corticosteroids or methotrexate . P01375 alpha inhibition as treatment modality for certain rheumatologic and gastrointestinal diseases . With the development of biologicals that specifically target tumor necrosis factor ( P01375 )alpha , our therapeutic approach to inflammatory diseases has dramatically changed . There are currently three anti-TNFalpha drugs available : etanercept , infliximab , and adalimumab . DB00005 is a recombinant fusion protein that can be used alone or in combination with other medications for conditions such as rheumatoid arthritis , juvenile rheumatoid arthritis , psoriatic arthritis , psoriasis , and ankylosing spondylitis . DB00065 , a chimeric humanized monoclonal antibody and adalimumab , a fully human monoclonal antibody are approved for the treatment of rheumatoid arthritis , psoriasis , psoriatic arthritis , ankylosing spondylitis , and moderate to severe Crohn 's disease . DB00065 is also approved for ulcerative colitis . Another anti-TNFalpha drug , certolizumab pegol , was declined approval as treatment option for active Crohn 's disease due to a lack of sufficient efficacy . Phase III studies for the treatment of rheumatoid arthritis patients are still pending . It is the goal of this review article to summarize various therapeutic indications , underlying studies , safety , and use during pregnancy , as well as future directions for anti- P01375 therapies . Inhibitors of P78536 and P29466 as anti-inflammatory drugs . P01375 neutralising agents such as DB00065 ( Remicade ) , DB00005 ( Enbrel ) and the IL-1 receptor antagonist DB00026 ( Kineret ) , are currently used clinically for the treatment of many inflammatory diseases such as Crohn 's disease , rheumatoid arthritis , ankylosing spondylitis , juvenile rheumatoid arthritis , psoriatic arthritis and psoriasis . These protein preparations are expensive to manufacture and administer , need to be injected and can cause allergic reactions . An alternative approach to lowering the levels of P01375 and IL-1beta in inflammatory disease , is to inhibit the enzymes that generate these cytokines using cheaper small molecules . This paper is a broad overview of the progress that has been achieved so far , with respect to small molecule inhibitor design and pharmacological studies ( in animals and humans ) , for the metalloprotease Tumour Necrosis Factor-alpha Converting Enzyme ( P78536 ) and the cysteine protease P29466 ( Interleukin-1beta Converting Enzyme , ICE ) . Inhibitors of these two enzymes are currently considered to be good therapeutic targets that have the potential to provide relatively inexpensive and orally bioavailable anti-inflammatory agents in the future . DB00065 and brucellosis : not the usual suspects , this time . P01375 ( P01375 ) plays an important role in the host defense mechanism , and anti- P01375 antibody therapies may increase the risk of serious infections . We herein report a case of 57-year-old male with rheumatoid arthritis who developed brucellosis during treatment with infliximab in combination with methotrexate and a low-dose steroid . Brucellosis should be kept in mind , particularly in endemic areas , in patients receiving anti- P01375 therapy . Clinicians should be aware of brucellosis symptoms and ways of contamination and should warn their patients . Early diagnosis and rapid treatment may prevent a possible poor course of the disease in immunocompromised patients . Efficacy , safety , and pharmacokinetics of multiple administration of infliximab in Behçet 's disease with refractory uveoretinitis . OBJECTIVE : Behçet 's disease ( BD ) with uveoretinitis is a chronic refractory disease accompanied by ocular attacks . As the decrease in visual acuity due to ocular attack is seriously life-threatening , development of a new drug is anticipated . Since tumor necrosis factor-a ( P01375 ) is involved in the symptoms of BD , particularly the activity of ocular symptoms , suppression of P01375 might be effective in treating BD with uveoretinitis . We conducted a clinical trial of infliximab , an anti- P01375 chimeric monoclonal antibody , in patients with BD . METHODS : In this open label trial , the efficacy , safety , and pharmacokinetics of repeated administration of infliximab were evaluated in 13 patients with BD accompanied by refractory uveoretinitis . DB00065 was administered 4 times at Weeks 0 , 2 , 6 , and 10 at doses of either 5 or 10 mg/kg by intravenous drip infusion . Frequency of ocular attacks was used as the primary index for evaluation of efficacy , with visual acuity and extraocular symptoms as secondary indices . RESULTS : The mean numbers of ocular attacks , converted to frequency per 14 weeks , were 3.96 times for the 5 mg/kg group and 3.79 times for the 10 mg/kg group during the observation period . Following treatment with infliximab , they decreased to 0.98 times and 0.16 times , respectively . A serious adverse event , tuberculosis , was observed in one case in the 10 mg/kg group . Serum infliximab concentration increased with dosage . CONCLUSION : Administration of infliximab in patients with BD with refractory uveoretinitis suppressed the frequency of ocular attacks , and multiple administration was well tolerated , suggesting that infliximab is effective for this condition . DB00065 monotherapy in psoriasis : a case of rapid clinical and histological response . BACKGROUND : Psoriasis is a common chronic relapsing , inflammatory , hyperproliferative skin disorder with genetic predisposition . There is currently no experimental model for psoriasis and the pathogenesis is not fully understood . Psoriatic plaques have been shown to contain increased levels of cytokines , including tumor necrosis factor alpha ( P01375 ) . Anti-tumor necrosis factor therapy with infliximab has been shown to be highly effective in recalcitrant psoriasis . METHODS : We evaluated the efficacy and timeline of histological changes in a psoriatic plaque following infliximab infusion . A patient with severe recalcitrant plaque psoriasis was clinically and histologically assessed for improvement . RESULTS : We found rapid clinical improvement with infliximab accompanied by histopathological changes . The earliest effects were seen on neutrophils and lymphocytes whereas keratinocyte normalization was not evident at the early stages . CONCLUSION : DB00065 is not only an effective agent in the treatment of psoriasis but appears to have a very rapid onset of action . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Role of anti-tumour necrosis factor-alpha therapeutic agents in the emergence of infections . There is increasing interest concerning the possible impact of anti-tumour necrosis factor ( P01375 ) -alpha therapeutic agents on the emergence of infections . However , these agents do not seem to increase the incidence of adverse infectious events significantly . Published observations concern mostly infections of the urinary and upper respiratory tracts that develop in the setting of co-morbidities , such as anterior or concomitant immunosuppressive treatment . DB00065 appears to increase the risk of tuberculosis , but this effect has not been observed with other anti- P01375 agents . To better characterise the adverse infectious effects associated with these agents , physicians should be encouraged to notify the microbiological data relating to all cases . Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus , expression of DB01221 receptor and inflammatory genes . Effects of C-phycocyanin ( C-PC ) , the active component of Spirulina platensis water extract on the expressions of N-methyl D-aspartate receptor subunit 2B ( Q13224 ) , tumor necrosis factor-α ( P01375 -α ) , interleukin-1β ( IL-1β ) , and cyclooxygenase type 2 ( P35354 ) genes in the cochlea and inferior colliculus ( IC ) of mice were evaluated after tinnitus was induced by intraperitoneal injection of salicylate . The results showed that 4-day salicylate treatment ( unlike 4-day saline treatment ) caused a significant increase in Q13224 , P01375 -α , and IL-1β mRNAs expression in the cochlea and IC . On the other hand , dietary supplementation with C-PC or Spirulina platensis water extract significantly reduced the salicylate-induced tinnitus and down-regulated the mRNAs expression of Q13224 , P01375 -α , IL-1β mRNAs , and P35354 genes in the cochlea and IC of mice . The changes of protein expression levels were generally correlated with those of mRNAs expression levels in the IC for above genes . Differential effects of decoy receptor- and antibody-mediated tumour necrosis factor blockage on FoxP3 expression in responsive arthritis patients . Our aim was to clarify if anti-tumour necrosis factor ( P01375 ) drugs have effect on expression of three splice forms of FoxP3 mRNA in blood P01730 + T cells from rheumatoid arthritis ( RA ) patients compared with healthy controls . Forty-five rheumatoid arthritis patients treated with anti- P01375 therapy were investigated in a 12-week prospective cohort study . FoxP3 isoforms , CD25 and P16410 mRNA in blood P01730 + T cells were measured with quantitative real-time PCR . Patients benefitting from the treatment , based on changes in DAS28 scores , revealed a significant decrease in expression of full-length FoxP3 following 12 weeks treatment with P01375 receptor 2 fusion protein ( DB00005 ) , but not following treatment with anti- P01375 antibodies ( DB00051 or DB00065 ) . A partial normalization of the P16410 /FoxP3fl ratio and a correlation between clinical improvement and change in FoxP3 mRNA expression were also seen in DB00005 responders . These changes were not observed in responsive patients treated with the antibody therapies . Our data suggest that P01375 decoy receptor and anti- P01375 antibodies differ in their effect on FoxP3 expression in responsive patients . As DB00005 binds both P01375 -α and Lymphotoxin-α ( LT-α ) , whereas the antibodies only target P01375 -α , LT-α may regulate FoxP3 expression in a subset of RA patients . Our findings support the view that anti- P01375 treatment is mainly symptomatic . P01375 blockade induce clinical remission in patients affected by polymyalgia rheumatica associated to diabetes mellitus and/or osteoporosis : a seven cases report . Polymyalgia rheumatica ( PMR ) is a chronic inflammatory condition of the elderly , characterized by aching and morning stiffness in the cervical region , shoulders and pelvic girdles . A steroid treatment course of 6-24 months is often required , but , due to important side effects , it is troublesome if the PMR patient is also affected by diabetes mellitus ( DM ) and/or osteoporosis . Aim of our study is to test anti- P01375 alpha treatment as a steroid sparing tool in PMR patients affected by DM or osteoporosis . In particular , we hypothesise that P01375 alpha blockade can be useful not only in remission maintaining , but also in the induction of clinical remission without corticosteroids in this kind of patients . In a six months follow up , patients had clinical improvement , confirmed by physical medical examination , and a statistically significant reduction in P03372 and CRP mean values . Anti- P01375 alpha treatment was well tolerated by all patients . These preliminary data suggest than DB00065 can be useful in the treatment of PMR patients , not only for steroid sparing purposes , but also as first line therapy in PMR patients with severe comorbidity , such as diabetes mellitus or osteoporosis . Anti-infliximab IgE and non-IgE antibodies and induction of infusion-related severe anaphylactic reactions . BACKGROUND : DB00065 is a chimeric monoclonal antibody against P01375 useful in the treatment of many chronic inflammatory diseases . Severe anaphylaxis has been reported during therapy , although the exact mechanism has not been fully defined . The reactions have been related to the infliximab immunogenicity and development of specific antibodies . AIMS OF THE STUDY : Evaluation of the development of IgE and non-IgE antibodies to infliximab and their relationship with infusion reaction . METHODS : Seventy-one patients ( 11 reactives , 11 therapeutically nonresponders , and 49 unreactive therapeutically responders ) and 20 non-infliximab-exposed control subjects ( ten rheumatoid arthritis , five spondyloarthropathies , five vasculitis ) were evaluated for the presence of IgE ( ImmunoCAP assay ) , IgM , and non-isotype-specific ( ELISA assays ) anti-infliximab antibodies . Sera were obtained at baseline and during the course of treatment , before each infliximab infusion . RESULTS : Eleven out of 71 patients had a hypersensitivity reaction to infliximab . Non-isotype-specific anti-infliximab antibodies were detected in eight reactive and two nonresponder patients . Three patients with severe reactions displayed anti-infliximab IgE antibodies and positive skin testing . Detectable levels of anti-infliximab IgM antibodies were shown in three additional IgE- and skin testing-negative patients . IgE and IgM antibodies to infliximab were not detectable in the two nonresponder patients . Antibodies developed before the 2nd and the 3rd infusion , and their appearance was strictly related to the timing of the reaction . CONCLUSIONS : This report indicates that in some patients with infliximab-related severe reactions , IgE or IgM antibodies against infliximab were detectable . The majority of reactions could be predicted by the appearance of anti-infliximab antibodies . Anti- P01375 and fistulizing perianal Crohn 's disease : use in clinical practice . Perianal fistulas are a major problem of patients with Crohn 's disease ( CD ) , and occur in up to 40 % of patients . The treatment of fistulizing perianal CD has recently largely evolved as a result of improvements of pharmacological and surgical approaches and the introduction of anti- P01375 treatment . Especially the use of anti- P01375 agents in complex or refractory perianal fistulas has been proven as the most effective medical treatment of this difficult to treat disease . DB00065 and adalimumab are the two currently available anti- P01375 agents that both have shown significant efficacy in the treatment and sustained remission of perianal fistulizing CD with comparable fistula closure rates . However , despite this treatment a large number of patients have continuous disease activity and high relapsing rates whereas only a small percentage of them have a complete fistula healing . Therefore the optimal outcome is still dependent on a multidisciplinary approach with a close interaction between gastroenterologists and surgeons . The individualised treatment based on anti- P01375 agents with the rational combination of antibiotic use , surgery and immunosuppressive therapy is , currently , the suggested treatment in order to achieve remission of a persistent perianal fistula . Large randomised studies are required for the long-term evaluation of the efficacy in modifying the disease course of this combined approach . DB00065 use in patients with severe graft-versus-host disease and other emerging risk factors of non-Candida invasive fungal infections in allogeneic hematopoietic stem cell transplant recipients : a cohort study . Acute graft-versus-host disease ( GVHD ) is a common complication of allogeneic hematopoietic stem cell transplantation ( HSCT ) . It has been proposed that tumor necrosis factor alpha ( P01375 ) blockade with infliximab may be an effective treatment for severe ( grades III-IV ) GVHD . We determined if infliximab use in this high-risk population was associated with an additional increased risk of non-Candida invasive fungal infections ( IFIs ) . Records of the 2000-2001 HSCT cohort at our institution were reviewed . Fifty-three ( 20 % ) of 264 evaluable patients developed severe GVHD and 11 of these 53 ( 21 % ) received infliximab for treatment . Proven or probable IFI was documented in 10 ( 19 % ) of 53 patients with severe GVHD ( incidence rate of 0.99 cases/1000 GVHD patient-days ) . When stratified by infliximab use , 5 of 11 infliximab recipients developed an IFI ( 6.78 cases/1000 GVHD patient-days ) , compared with 5 of 42 IFI cases among nonrecipients ( 0.53 cases/1000 GVHD patient-days ) . In a time-dependent Cox regression model among patients with severe GVHD , the adjusted IFI hazard ratio of infliximab exposure was 13.6 ( P =.004 ; 95 % CI , 2.29-80.2 ) . We conclude that infliximab administration is associated with a significantly increased risk of non-Candida IFI in HSCT recipients with severe GVHD disease . Pre-emptive systemic antifungal therapy against molds should be considered in patients who develop severe GVHD after HSCT if infliximab is used . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . A SPECIAL MEETING REVIEW EDITION : Highlights in Anti-Tumor Necrosis Factor Monitoring and Antibody Monitoring From the 2014 DDW Meeting : Digestive Disease Week 2014 May 3-6 , 2014 • Chicago , Illinois : Special Reporting on:• Therapeutic Monitoring of Anti- P01375 Levels and Antibodies to Predict Response and Achieve Mucosal Healing• Prospective Therapeutic Drug Monitoring and Optimization of DB00065 Maintenance Therapy in IBD• Classification of Non- Q9UKU7 , Crohn 's Disease and Ulcerative Colitis in a Young Patient Population Using a Multi-Marker Diagnostic Panel• Persistence of Antibodies to DB00065 for More Than Two Months Predicts Loss of Response to DB00065 in Inflammatory Bowel Diseases• Pre-Operative Serological Markers May Predict Postoperative Crohn 's Disease Recurrence : Results From a Prospective Mono-Centric Trial• Antibodies and Levels of Biologies-Reactive vs Proactive Measurements• Higher DB00352 Nucleotide Concentrations Are Associated With Higher Trough Levels of DB00065 in Patients on Combination Therapy• The Clinical and Immunological Significance of Low Levels of DB00065 in the Absence of Anti-lnfliximab Antibodies in Patients With IBD• Antibodies to DB00051 Predict Inflammation in Crohn 's Patients on Maintenance DB00051 Therapy• Q676U5 Genotype Is Associated With Response to Anti-TNFWith Expert Commentary by:William J. Sandborn , MDProfessor and Chief , Division of Gastroenterology Director , UCSD Q9UKU7 CenterUC San Diego Health SystemLa Jolla , California . Ischaemic/reperfusion injury : Role of infliximab . Ischaemia/reperfusion ( I/R ) injury is an underlying complex interrelated patho-physiological process which effects the outcome of many clinical situations , in particular transplantation . P01375 ( P01375 ) -α is a pleiotropic inflammatory cytokine ; a trimeric protein encoded within the major histocompatibility complex which plays a pivotal role in this disease process . This review is based at looking into an update , particularly the new insights in the mechanisms of action of P01375 antagonist such as infliximab . DB00065 may thus play a dual role in the field of transplantation where it might not only down regulate the I/R injury , it may also have a beneficial role in the reduction of acute rejection . Anti-allergic effects of nilotinib on mast cell-mediated anaphylaxis like reactions . DB04868 is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of P11274 - P00519 -positive chronic myelogenous leukemia . However , its effect on mast cell-mediated anaphylactic reaction is still not known . The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action . DB04868 administration prevented systemic anaphylaxis in mice , mediated by compound 48/80 , in a dose- and time-dependent manner . Also , nilotinib significantly inhibited ( P < 0.05 ) allergic paw edema in rats . Furthermore , nilotinib significantly decreased ( P < 0.05 ) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner . In addition , nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin . Moreover , nilotinib attenuated the secretion of pro-inflammatory cytokine , tumor necrosis factor ( P01375 ) -α expression in the rat peritoneal mast cells . These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . DB00065 ( Remicade ) in the treatment of psoriatic arthritis . Elucidation of the cellular immunopathology and cytokine profile of psoriatic arthritis ( PsA ) , a chronic inflammatory disease associated with psoriasis , has resulted in the development of a number of novel biologic therapies . Among these biologics , tumor necrosis factor-alpha ( P01375 ) inhibitors have been used successfully to treat patients suffering from rheumatoid arthritis or psoriasis . The pivotal role of P01375 in the pathogenesis and progression of PsA suggested that anti- P01375 agents could be effective in controlling PsA . The results from two large , randomized , double-blind , placebo-controlled trials in patients with moderate to severe PsA indicated that the anti- P01375 -inhibitor , infliximab , can control both the joint and skin manifestations of the disease . This review focuses on the clinical development of infliximab as a treatment for PsA . The development of other anti- P01375 biologics is also discussed . Protective effects of indomethacin and cyclophosphamide but not of infliximab on liver metabolic changes caused by adjuvant-induced arthritis . In the study , indomethacin , cyclophosphamide , and infliximab were administered to adjuvant-induced arthritic rats to determine if they were able to prevent the abnormalities caused by arthritis on hepatic metabolism . The drugs were administered to arthritic rats , and at the clinical onset of arthritis ( day 14 after adjuvant injection ) , the livers were perfused to evaluate gluconeogenesis , ureagenesis , oxygen uptake , L : -lactate , pyruvate , and ammonia release from L : -alanine . The effects of the drugs on body weight gain and the signs of arthritis ( paw edema , appearance of secondary lesions , and weights of lymphoid tissues ) were also evaluated . Cyclophosphamide could completely prevent liver metabolic changes and the inflammatory response . Indomethacin restored ureagenesis , minimized the decrease in gluconeogenesis , and exerted a partially beneficial effect on inflammatory reactions . DB00065 did not improve arthritis-induced liver metabolic alterations or inflammatory responses . These results suggest the participation of prostaglandins , but not P01375 -α , on arthritis-induced liver metabolic alterations . DB00065 inhibits bone resorption by circulating osteoclast precursor cells in patients with rheumatoid arthritis and ankylosing spondylitis . OBJECTIVE : To examine the effects of infliximab on bone resorption by osteoclast precursor cells ( OCPs ) in patients with rheumatoid arthritis ( RA ) and ankylosing spondylitis ( AS ) and to compare the results with changes in disease activity . METHODS : Before and during 24 weeks of infliximab treatment , peripheral blood mononuclear cells of 9 RA and 10 AS patients were seeded onto ivory wafers and adherent cells , including OCPs , were grown in medium promoting osteoclast differentiation . Bone resorption was evaluated morphometrically and correlated to disease activity . A total of 19 healthy individuals were studied in parallel . In addition , biochemical bone markers were assessed in all patients at baseline and after 24 weeks . RESULTS : OCPs from RA patients showed a higher bone resorption at baseline when compared to AS patients . Blocking of tumour necrosis factor ( P01375 )alpha with infliximab resulted in a strong reduction of bone resorption by OCPs in both cohorts and occurred faster in RA compared to AS patients . This inhibition coincided with a reduction of clinical disease activity in both patient cohorts and with an increase of serum osteocalcin levels and a relative decrease of collagen crosslinks in RA compared to AS patients . CONCLUSION : These results provide an explanation on the cellular level for the anticatabolic effect of P01375 neutralisation on bone . The variation in the kinetics of bone resorption by the OCPs in patients with RA and AS suggests disease-specific differences in the type or in the preactivation of OCPs . Does weight-adjusted anti-tumour necrosis factor treatment favour obese patients with Crohn 's disease ? BACKGROUND : DB00051 ( P00813 ) is a subcutaneous anti-tumour necrosis factor ( anti- P01375 ) agent , effective in inducing and maintaining remission in Crohn 's disease ( CD ) . Unlike DB00065 ( IFX ) , P00813 dosing is not weight adjusted and dose frequency is based on clinical response . AIM : To determine whether obesity is a risk factor for early loss of response ( P23490 ) to anti- P01375 treatment and whether weight-adjusted anti- P01375 treatment is favourable . MATERIALS AND METHODS : A hospital database of CD patients receiving anti- P01375 treatment was analyzed retrospectively . The relationship between time to P23490 and BMI was examined by Kaplan-Meier ( KM ) survival curves and a Cox proportional hazards model . RESULTS : P00813 patients : Of the 54 patients ( 46 BMI < 30 and 8 BMI≥30 ) , KM estimation indicated a significantly shorter time to dose escalation in the BMI of at least 30 ( χ=6.117 , P=0.01 ) . The Cox proportional hazards model showed that an increased hazard of P23490 to P00813 is related to increases in BMI ( P=0.04 ) . IFX patients : Of the 76 patients ( 62 BMI < 30 and 14 BMI≥30 ) , KM estimation showed that the differences in survival curves were not significant ( χ=1.933 , P=0.16 ) for the BMI groups . This was supported by the Cox proportional hazard model ( P=0.36 ) . CONCLUSION : BMI appears to be important in predicting P00813 efficacy ( P23490 ) in CD . IFX appears to overcome this reduction of efficacy in obese patients . A prospective study evaluating the effect of weight on anti- P01375 drug response and serum drug levels is warranted . Effect of DB00065 on the UVB-Induced Apoptosis of Keratinocytes Infected by HPV38 E6/E7 . The question of the effect of anti- P01375 in skin carcinogenesis is especially relevant in view of the increased use of these drugs for the treatment of autoinflammatory immune diseases . Since ultraviolet radiation and human papillomavirus are involved in skin carcinogenesis , we wished to investigate the effect of P01375 antagonists on the UVB-induced apoptosis of keratinocytes infected by HPV38 . Our results indicate that anti- P01375 agent , infliximab , does not contribute to the survival of HPV38-transduced keratinocytes with UVB-induced DNA damages . Q9GZV9 contributes to diminished bone mineral density in childhood inflammatory bowel disease . BACKGROUND : Diminished bone mineral density ( BMD ) is of significant concern in pediatric inflammatory bowel disease ( Q9UKU7 ) . Exact etiology is debatable . The recognition of fibroblast growth factor 23 ( Q9GZV9 ) , a phosphaturic hormone related to tumor necrosis factor alpha ( P01375 -α ) makes it plausible to hypothesize its possible relation to this pathology . METHODS : In this follow up case control study , BMD as well as serum levels of Q9GZV9 , calcium , phosphorus , alkaline phosphatase , creatinine , parathyroid hormone , 25 hydroxy vitamin D3 and 1 , 25 dihydroxy vitamin D3 were measured in 47 children with Q9UKU7 during flare and reassessed in the next remission . RESULTS : Low BMD was frequent during Q9UKU7 flare ( 87.2 % ) with significant improvement after remission ( 44.7 % ) . During disease flare , only 21.3 % of patients had vitamin D deficiency , which was severe in 12.8 % . During remission , all patients had normal vitamin D except for two patients with Crohn 's disease ( CD ) who remained vitamin D deficient . Mean value of serum Q9GZV9 was significantly higher among patients with Q9UKU7 during flare compared to controls . It showed significant improvement during remission but not to the control values . 1 , 25 dihydroxy vitamin D3 , Q9GZV9 , serum calcium and urinary phosphorus were significant determinants of BMD in Q9UKU7 patients . CONCLUSIONS : We can conclude that diminished BMD in childhood Q9UKU7 is a common multifactorial problem . Elevated Q9GZV9 would be a novel addition to the list of factors affecting bone mineral density in this context . Further molecular studies are warranted to display the exact interplay of these factors . Autosomal-dominant hypophosphatemic rickets ( P30518 ) mutations stabilize Q9GZV9 . BACKGROUND : The gene for the renal phosphate wasting disorder autosomal-dominant hypophosphatemic rickets ( P30518 ) is Q9GZV9 , which encodes a secreted protein related to the fibroblast growth factors ( FGFs ) . We previously detected missense mutations R176Q , R179W , and R179Q in Q9GZV9 from P30518 kindreds . The mutations replace R residues within a subtilisin-like proprotein convertase ( Q969E3 ) cleavage site 176RHTR-179 ( RXXR motif ) . The goal of these studies was to determine if the P30518 mutations lead to protease resistance of Q9GZV9 . METHODS : The P30518 mutations were introduced into human Q9GZV9 cDNA clones with or without an N-terminal FLAG tag by site-directed mutagenesis and were transiently transfected into HEK293 cells . Protein expression was determined by Western analyses . RESULTS : Antibodies directed toward the C-terminal portion of Q9GZV9 revealed that the native Q9GZV9 protein resolved as 32 kD and 12 kD species in HEK293 conditioned media ; however , the three mutated proteins were detected only as the 32 kD band . An N-terminal FLAG-tagged native Q9GZV9 resolved as two bands of 36 kD and 26 kD when detected with a FLAG antibody , whereas the R176Q mutant resolved primarily as the 36 kD protein species . Cleavage of Q9GZV9 was not enhanced by extracellular incubation of Q9GZV9 with HEK293 cells . Native and mutant FGF-23s bound heparin . CONCLUSIONS : Q9GZV9 proteins containing the P30518 mutations are secreted , and produce polypeptides less sensitive to protease cleavage than wild-type Q9GZV9 . Therefore , the P30518 mutations may protect Q9GZV9 from proteolysis , thereby potentially elevating circulating concentrations of Q9GZV9 and leading to phosphate wasting in P30518 patients . Delayed addition of tumor necrosis factor ( P01375 ) antagonists inhibits the generation of CD11c+ dendritic cells derived from P28906 + cells exposed to P01375 . We have developed a method that cells exhibiting typical dendritic cell ( DC ) characteristics are generated from human P28906 (+) cells and phagocytose cogenerating erythroid progenitor cells in the presence of tumor necrosis factor-alpha ( P01375 ) , interleukin-3 , stem cell factor and erythropoietin . Using this system , we titrated the effects of P01375 antagonists , etanercept and infliximab , on P01375 activity . We found that 1 microg/ml etanercept dramatically inhibited the generation of CD11c(+) cells accompanying with a complete recovery of the generation of erythroid progenitors . DB00065 at 200 microg/ml exhibited a similar effect to that observed for etanercept . The delayed addition of etanercept to this culture system at day five resulted in significant inhibitory effects on the generation of CD11c(+) , P01730 (+) and P42081 (+) cells . These results indicate that P01375 antagonists administered at a concentration that is achievable in vivo , neutralize the biologic effects of P01375 in generating CD11c(+) cells and that a delay in the administration of these antagonists for as long as 5 days partially inhibits the biologic activity of P01375 . These findings may contribute to a great understanding of anti- P01375 therapy in patients with an overproduction of cytokines such as hemophagocytic syndromes . DB00065 therapy in pulmonary fibrosis associated with collagen vascular disease . OBJECTIVE : To study the potential effectiveness of tumor necrosis factor a ( P01375 ) inhibitor treatment for pulmonary fibrosis associated with a collagen vascular disease , CVD ( rheumatoid arthritis , RA and systemic sclerosis , SSc ) refractory to conventional treatment . METHODS : Four patients ( three men with RA , one woman with SSc ) were treated with infliximab . All patients received 3mg/kgr of infliximab at intervals 0 , 2 and 6 weeks , and then maintenance infusions every 8 weeks afterwards for at least a 12-month period . Patients had active disease despite treatment with corticosteroids and other immunomodulatory agents . RESULTS : Treatment was well-tolerated from all patients . Pulmonary fibrosis remained stable during treatment in terms of symptoms , pulmonary function tests ( PFTs ) and High resolution computed tomography ( HRCT ) appearance . As expected , a clinical response was observed in joint symptoms in patients with RA as evaluated by the DAS28 ( Disease Activity Score , the 28 joint version ) . CONCLUSION : This study suggests that inhibition of P01375 with infliximab may stabilize the progression of pulmonary fibrosis associated with CVD . Prospective , controlled trials are necessary to determine the efficacy of infliximab in pulmonary fibrosis associated CVD . Treatment with anti-tumor necrosis factor alpha ( P01375 ) monoclonal antibody dramatically decreases the clinical activity of psoriasis lesions . We treated a 57-year-old woman for refractory inflammatory bowel disease with a humanized anti-tumor necrosis factor alpha monoclonal antibody ( DB00065 ) . The patient also had a 15-year history of Crohn 's disease and a 20-year history of moderate to severe psoriasis . She received a single infusion of DB00065 ( 5 mg/kg ) . Two weeks after the infusion her psoriasis had dramatically improved in appearance . To our knowledge , our case is the first reported instance of successful anti-tumor necrosis factor alpha therapy in psoriasis . Comparison of drug survival rates for tumor necrosis factor antagonists in rheumatoid arthritis . BACKGROUND : Persistence of anti-tumor necrosis factor ( P01375 ) therapy in rheumatoid arthritis ( RA ) is an overall marker of treatment success . OBJECTIVE : To assess the survival of anti- P01375 treatment and to define the potential predictors of drug discontinuation in RA , in order to verify the adequacy of current practices . DESIGN : An observational , descriptive , longitudinal , retrospective study . SETTING : The Hospital Clínico Universitario de Valladolid , Valladolid , Spain . PATIENTS : RA patients treated with anti- P01375 therapy between January 2011 and January 2012 . MEASUREMENTS : Demographic information and therapy assessments were gathered from medical and pharmaceutical records . Data is expressed as means ( standard deviations ) for quantitative variables and frequency distribution for qualitative variables . Kaplan-Meier survival analysis was used to assess persistence , and Cox multivariate regression models were used to assess potential predictors of treatment discontinuation . RESULTS : In total , 126 treatment series with infliximab ( n = 53 ) , etanercept ( n = 51 ) or adalimumab ( n = 22 ) were administered to 91 patients . DB00065 has mostly been used as a first-line treatment , but it was the drug with the shortest time until a change of treatment . Significant predictors of drug survival were : age ; the anti- P01375 agent ; and the previous response to an anti- P01375 drug . LIMITATION : The small sample size . CONCLUSION : The overall efficacy of anti- P01375 drugs diminishes with time , with infliximab having the shortest time until a change of treatment . The management of biologic therapy in patients with RA should be reconsidered in order to achieve disease control with a reduction in costs . Chronic administration of infliximab ( P01375 -α inhibitor ) decreases depression and anxiety-like behaviour in rat model of chronic mild stress . Pro-inflammatory cytokines have been proposed to be associated with the pathogenesis of depression . Consistent with this notion , several clinical observations have suggested the antidepressant efficacy of P01375 -α inhibitors in patients with chronic inflammatory diseases . In this study , we evaluated the antidepressant and anxiolytic effects of chronic P01375 -α inhibitor ( infliximab , 5 mg/kg , i.p. , weekly ) administration in the chronic mild stress ( CMS ) model of depression . Rats were divided into three groups : saline-control ( no stress ) , saline-CMS , and infliximab-CMS . Rats in the latter two groups were exposed to CMS for 8 weeks . Saline ( former two groups ) or infliximab was injected weekly during this period . After CMS , total locomotor activity , anxiety-like behaviour and depression-like behaviours were evaluated using automated locomotor activity cage , elevated plus maze ( EPM ) , and sucrose preference ( P21549 ) and forced swimming ( FS ) tests , respectively . As expected , the saline-CMS group exhibited higher depression-like behaviours in FS and P21549 tests compared with the saline-control group . There were no differences between these two groups in terms of the anxiety-like behaviour or total locomotor activity . DB00065 reduced the depression-like behaviour of CMS rats compared with saline-CMS group , and anxiety-like behaviour of CMS rats compared with saline-CMS and saline-control groups . Our findings suggest that chronic and systemic P01375 -α inhibition reduced depression and anxiety-like behaviour in the CMS model of depression in rats . Effect of the anti-tumor necrosis factor-alpha antibody infliximab on the ex vivo mucosal matrix metalloproteinase-proteolytic phenotype in inflammatory bowel disease . BACKGROUND : Previous studies have shown an upregulation of matrix metalloproteinases ( MMPs ) in intestinal tissue of patients with inflammatory bowel disease ( Q9UKU7 ) and significant clinical improvement after administration of the anti- P01375 antibody infliximab . The aims of our study were to determine expression and secretion of P03956 , -2 , -3 , -9 , and their inhibitors P01033 , -2 by Q9UKU7 versus control intestinal mucosa ex vivo and to assess the regulatory capacity by infliximab of the proteolytic phenotype . METHODS : Intestinal mucosal explants from 20 Q9UKU7 and 15 control patients were cultured with or without infliximab and/or the T-cell activator pokeweed mitogen ( PWM ) . Explants and culture supernatants were analyzed for MMPs , TIMPs , and P01375 protein , activity and/or mRNA levels . All patients were genotyped for functional P01375 , MMP , and P01033 single nucleotide polymorphism ( SNP ) loci . RESULTS : Expression of MMP and P01033 protein/activity in basal medium was higher in Q9UKU7 versus control explants . Dependent on genotype at SNP loci , infliximab downregulated P03956 , -3 , and -9 relative to P01033 and -2 and also decreased P03956 and -3 activities , while PWM enhanced these levels , partly counteracted again by infliximab . The expression of P08253 relative to P01033 did not change by treatment with infliximab and/or PWM . CONCLUSIONS : The high expression of MMPs in patients with Q9UKU7 suggests a role for these proteinases in the pathogenesis of this disease . DB00065 seems to induce a genotype-associated matrix protective phenotype , which may contribute to the observed therapeutic efficacy of this drug in Q9UKU7 , particularly at the mucosal surface . Outcome of pregnancy in women with inflammatory bowel disease treated with antitumor necrosis factor therapy . BACKGROUND : DB00065 ( IFX ) and adalimumab ( P00813 ) are attractive treatment options in patients with inflammatory bowel disease ( Q9UKU7 ) also during pregnancy but there is still limited data on the benefit/risk profile of IFX and P00813 during pregnancy . METHODS : This observational study assessed pregnancy outcomes in 212 women with Q9UKU7 under antitumor necrosis factor alpha ( P01375 ) treatment at our Q9UKU7 unit . Pregnancy outcomes in 42 pregnancies with direct exposure to anti- P01375 treatment ( 35 IFX , 7 P00813 ) were compared with that in 23 pregnancies prior to Q9UKU7 diagnosis , 78 pregnancies before start of IFX , 53 pregnancies with indirect exposure to IFX , and 56 matched pregnancies in healthy women . RESULTS : Thirty-two of the 42 pregnancies ended in live births with a median gestational age of 38 weeks ( interquartile range [ IQR ] 37-39 ) . There were seven premature deliveries , six children had low birth weight , and there was one stillbirth . One boy weighed 1640 g delivered at week 33 , died at age of 13 days because of necrotizing enterocolitis . A total of eight abortions ( one patient wish ) occurred in seven women . Trisomy 18 was diagnosed in one fetus of a mother with CD at age 37 under P00813 treatment ( 40 mg weekly ) and pregnancy was terminated . Pregnancy outcomes after direct exposure to anti- P01375 treatment were not different from those in pregnancies before anti- P01375 treatment or with indirect exposure to anti- P01375 treatment but outcomes were worse than in pregnancies before Q9UKU7 diagnosis . CONCLUSIONS : Direct exposure to anti- P01375 treatment during pregnancy was not related to a higher incidence of adverse pregnancy outcomes than Q9UKU7 overall . Catecholamine-producing cells in the synovial tissue during arthritis : modulation of sympathetic neurotransmitters as new therapeutic target . BACKGROUND : The proinflammatory and anti-inflammatory role of the sympathetic nervous system in early and late inflammation is an unresolved paradox . A drastic loss of sympathetic nerve fibres in the synovial tissue of patients with rheumatoid arthritis ( RA ) has previously been demonstrated . The presence of tyrosine hydroxylase ( TH ) -positive cells in RA and osteoarthritis ( OA ) has been determined , but the role of these cells in inflammation is still unclear . OBJECTIVE : To characterise TH-positive cells in inflamed RA and OA synovial tissue and to study their role in inflammation . METHODS : Synovial samples were obtained from 32 patients with OA and 19 patients with RA and from 10 control patients . Synovial tissue samples were used for immunofluorescence staining . Synovial cells were isolated by tissue digestion and immediately used for cell culture . For in vivo experiments , collagen type-II arthritis in DBA/1J mice was induced . RESULTS : TH+ cells were present only in inflamed tissue and not in controls . Catecholamine-storing vesicles and vesicular monoamine transporter 2 ( Q05940 ) were identified in the synovial tissue . Experimental increase of cytoplasmic catecholamines by Q05940 blockade strongly reduced tumour necrosis factor ( P01375 ) independently of canonical extracellular β-adrenergic signalling . In addition , Q05940 blockade increased cyclic AMP ( DB02527 ) and DB02527 responsive element binding protein , responsible for P01375 inhibition . In vivo , appearance of Q05940 positive cells was confirmed . Q05940 blockade ameliorated inflammation also in vivo . CONCLUSIONS : This study demonstrates that local catecholamine-producing cells start to replace sympathetic nerve fibres around the onset of disease , and modulation of locally produced catecholamines has strong anti-inflammatory effects in vivo and in vitro .

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The effects of P08514 -IIIa antagonists and a combination of three other antiplatelet agents on platelet-leukocyte interactions . The effects of the P08514 -IIIa antagonists abciximab and MK-852 on platelet-leukocyte interactions in vitro were studied and the results compared with those obtained with a combination of aspirin , dipyridamole and AR-C69931 ( DB00128 /Dip/AR-C ) . Platelet-monocyte ( P/M ) and platelet-neutrophil ( P/N ) conjugate formation increased when blood was stirred or a platelet agonist was added . Leukocyte activation also occurred as judged by expression of surface tissue factor antigen and CD11b . DB00054 and MK-852 potentiated P/M , especially when collagen was used . They also increased the amount of tissue factor on the monocytes , but not CD11b . The DB00128 /Dip/AR-C did not enhance P/M or tissue factor exposure . Augmented tissue factor expression on monocytes in the presence of a P08514 -IIIa antagonist may be relevant to the increased mortality associated with trials of such antagonists when given orally in patients with vascular disease . The DB00128 /Dip/AR-C was superior to abciximab and MK-852 in inhibiting platelet and leukocyte function . A chimeric murine/human antibody Fab fragment directed against the platelet P08514 /IIIa receptor enhances and sustains arterial thrombolysis with recombinant tissue-type plasminogen activator in baboons . Inhibition of the platelet glycoprotein ( GP ) IIb/IIIa receptor with the murine monoclonal antibody 7E3 abolishes ex vivo platelet aggregation , reduces thrombogenicity , and sustains arterial recanalization with recombinant tissue-type plasminogen activator ( rt-PA ) . A chimeric murine/human Fab fragment of DB00054 ( c7E3-Fab ) has a markedly reduced immunogenicity , but its potency as an adjunct for thrombolysis with rt-PA has not been evaluated . The effects of a single intravenous bolus injection of aspirin ( 17 mg/kg ) or c7E3-Fab ( 0.45 mg/kg ) on thrombolysis and reocclusion induced with rt-PA were studied in groups of six baboons with femoral arterial thrombosis and superimposed high-grade stenosis . This dose of c7E3-Fab blocked 96 +/- 1 % of the platelet P08514 /IIIa receptors and abolished ADP-induced platelet aggregation . Bolus intravenous injections of rt-PA ( 0.25 mg/kg ) were repeated at 15-minute intervals until reperfusion occurred ( maximum of four injections ) . In the aspirin group , reperfusion was obtained within 51 +/- 16 minutes ( mean +/- SD ) but was rapidly followed by reocclusion within 6 +/- 9 minutes and by cyclic reflow and reocclusion . In the c7E3-Fab group , reperfusion was obtained within 25 +/- 8 minutes ( P < .01 versus aspirin group ) and was associated with a delayed reocclusion of 63 +/- 63 minutes ( P < .05 versus aspirin group ) . Template bleeding times remained unchanged in the aspirin/rt-PA group but were markedly prolonged ( to > 30 minutes ) in the c7E3-Fab/rt-PA group. ( ABSTRACT TRUNCATED AT 250 WORDS ) Comparative studies of a humanized anti-glycoprotein IIb/IIIa monoclonal antibody , YM337 , and abciximab on in vitro antiplatelet effect and binding properties . The effects of YM337 , the Fab fragment of a humanized anti-glycoprotein IIb/IIIa ( P08514 /IIIa ) monoclonal antibody C4G1 , on in vitro platelet function and binding properties were compared with those of abciximab , the Fab fragment of the human/murine chimeric anti- P08514 /IIIa monoclonal antibody DB00054 . Both agents completely inhibited platelet aggregation caused by all agonists tested except ristocetin . Further , both inhibited human platelet adhesion to P04275 , fibrinogen , fibronectin and subendothelial matrix with similar potency . DB09222 binding to washed platelets was dose-dependently inhibited by both agents . In binding assay using 125I-YM337 and 125I-abciximab , Kd values determined with platelet-rich plasma were 6.74 +/- 0.56 nM for YM337 and 6.65 +/- 1.45 nM for abciximab , and the number of binding sites were 42,700 +/- 3,000 for YM337 and 76,000 +/- 5,400 for abciximab . P08514 /IIIa was precipitated from the solubilized fraction of platelets by both agents . In contrast , integrin alphavbeta3 was precipitated from the solubilized fraction of human umbilical vein endothelial cells by abciximab but not by YM337 . DB09222 binding to purified P08514 /IIIa was dose-dependently inhibited by both agents . In contrast , vitronectin binding to purified integrin alphavbeta3 was dose-dependently inhibited by abciximab but not by YM337 , supporting the idea that abciximab reacts to integrin alphavbeta3 . Therefore , YM337 was suggested to bind to a different epitope of P08514 /IIIa from abciximab . These results suggest that YM337 specifically acts on platelet P08514 /IIIa receptors and has similar inhibitory properties on platelet aggregation and platelet adhesion to abciximab . Factor XIIIa binding to activated platelets is mediated through activation of glycoprotein IIb-IIIa . Stabilization of a clot is dependent on fibrin cross-linking mediated by the transglutaminase , factor XIIIa ( FXIIIa ) . In addition to fibrin stabilization , FXIIIa acts on a number of platelet-reactive proteins , including fibronectin and vitronectin , as well as the platelet proteins , glycoprotein ( GP ) IIb-IIIa , myosin , and actin . However , conditions inducing the platelet-activation dependent binding of FXIIIa have not been characterized nor have the sites mediating FXIIIa binding been identified . The generation of FXIIIa and consequent detection of FXIIIa on the platelet surface were compared with other thrombin-induced activation events ; the rate at which FXIIIa bound to activated platelets was much slower than platelet degranulation or fibrin(ogen) binding . Whereas platelets could be rapidly induced to express a functional receptor for FXIIIa , the rate of FXIIIa binding to platelets is limited by the rate of conversion of FXIII to FXIIIa . Immunoprecipitation of radiolabeled platelets using polyclonal anti-FXIII A-chain antibody identified two proteins corresponding to P08514 and P05106 . Preincubation of intact platelets with DB00054 , a monoclonal antibody that blocks the fibrinogen binding site , or GRGDSP peptide inhibited FXIIIa binding by about 95 % when measured by flow cytometry ; FXIIIa binding to purified P08514 -IIIa was also inhibited by DB00054 . The binding of FXIIIa to purified P08514 -IIIa was enhanced by the addition of fibrinogen , but not by that of fibronectin or thrombospondin , suggesting that FXIIIa also binds to fibrinogen associated with the complex . These observations suggest that activated platelets bearing FXIIIa may enhance stabilization of platelet-rich thrombi through surface-localized cross-linking events . Genetic aspects of ischemic stroke : coagulation , homocysteine , and lipoprotein metabolism as potential risk factors . Stroke is one of the most common causes of death and long term disability throughout the world . It may be the outcome of a number of monogenic disorders or , more commonly , a polygenic multifactorial disease . Numerous studies have investigated the role of genetics in the pathogenesis of ischemic stroke , with varied and often contradictory results . The candidate ' stroke risk ' genes affecting haemostasis ( P12259 , F2 , P02671 / P02675 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / P08514 , P17301 , P07359 , TPA , Q96IY4 , P07204 , PZ , P08758 ) , homocysteine metabolism ( P42898 , P35520 , Q99707 ) , and lipid metabolism ( apo E , P06858 , P11597 , O95477 , apo AI , apo CIII , apo AIV , apo AV , apo B , apo H , apo(a) , P27169 /2/3 , P01130 / P78380 ) are evaluated in this review . By examining meta-analyses and case-control studies , we made a classification of gene/gene polymorphisms according to the degree of association with ischemic stroke risk . The data assembled could be very useful for further meta-analysis and for future clinical applications . Signal transduction by HLA class II molecules in human T cells : induction of LFA-1-dependent and independent adhesion . Crosslinking HLA-DR molecules by monoclonal antibodies ( moAbs ) induces protein tyrosine phosphorylation and results in a secondary elevation of free cytoplasmic calcium concentrations in activated human T cells . Binding of bacterial superantigens or moAbs to DR molecules on activated T cells was recently reported to induce homotypic aggregation through activation of protein kinase C ( PKC ) and mediated by CD11a/CD54 ( LFA-1/ P62158 -1 ) adhesion molecules . Here , we report that moAbs directed against framework DR , but neither DR1 , 2- and DRw52- nor DQ- and DP-specific moABs induced homotypic aggregation of antigen- and alloantigen-activated T cells , antigen-specific P01730 + T-cell lines , a CD8+ T-cytotoxic cell line , and T-leukemia cells ( HUT78 ) . Protein tyrosine kinase ( PTK ) inhibitor herbimycin A partly blocked class-II-induced aggregation responses . In contrast , phorbol ester ( PMA ) -induced aggregation was essentially unaffected . A potent inhibitor of PKC , staurosporin , inhibited both moAb- and PMA-induced aggregation responses . The aggregation responses were completely inhibited by low temperatures , cytochalasins B and E , and partly inhibited by DB00974 and P05107 moAbs , but unaffected by aphidicolin , mitomycin C , an adenylate cyclase inhibitor ( 2'5'-dideoxyadenosine ) , and moAbs against other adhesion molecules ( P06729 / P19256 [ LFA-3 ] , P10747 / P10747 ligand P33681 , P01730 , and P16070 ) . In conclusion , HLA class-II-induced aggregation responses in activated T cells appear to involve PTK and PKC activation and to be mediated through CD11a-dependent and independent adhesion pathways . Combined thrombophilic polymorphisms in women with idiopathic recurrent miscarriage . OBJECTIVE : To identify associations or interrelations between carriage of the methylenetetrahydrofolate reductase ( P42898 ) C677T , the P42898 A1298C , the factor V Leiden G1691A , the factor II prothrombin G20210A , the human platelet antigen ( Q9Y251 ) 1 C12548T , and the apolipoprotein ( APO ) B R3500Q polymorphisms and idiopathic recurrent miscarriage ( IRM ) . DESIGN : Prospective case control study . SETTING : Academic research institution . PATIENT(S) : One hundred forty-five women with a history of three or more consecutive pregnancy losses before 20 weeks gestation and 101 healthy postmenopausal women with at least two live births and no history of pregnancy loss . INTERVENTION(S) : Peripheral venous punctures . MAIN OUTCOME MEASURE(S) : Multiplex polymerase chain reaction was performed to identify the different alleles of six candidate genetic risk factors for IRM ( P42898 C677T , P42898 A1298C , factor V Leiden G1691A , factor II prothrombin G20210A , Q9Y251 1 C12548T , and the APO B R3500Q ) . RESULT(S) : Allele and genotype frequencies of all polymorphisms were not significantly different between the study and the control groups . Also , no significant associations occurred between combinations of polymorphisms and the occurrence of IRM . CONCLUSION(S) : Our data fall short of showing any significant association between single polymorphisms of the P42898 , the Factor V Leiden , the Factor II P00734 , the Q9Y251 1 and APO B genes or combinations of these polymorphisms and the occurrence of IRM . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Forced expression of P38936 in P08514 - P38936 transgenic mice induces abnormalities in the proliferation of erythroid and megakaryocyte progenitors and primitive hematopoietic cells . OBJECTIVE : P38936 ( P38936 /Cip/kip) and p27(Kip1) are cyclin-dependant kinase inhibitors controlling cell-cycle exit and differentiation of numerous cell types . Among hematopoietic cells , megakaryocytes express high levels of P38936 , while in erythroid cells , p27(Kip1) is predominant . As P38936 and p27 could display overlapping functions and as megakaryocytes and erythroid cells derive from a bipotent progenitor , we developed an in vivo model to determine the specific role of P38936 in controlling the proliferation/differentiation balance of erythroid and megakaryocytic progenitors . METHODS : Transgenic mice that overexpressed P38936 under the control of the human P08514 promoter in early progenitors and along megakaryocytic differentiation were generated . Different subsets of hematopoietic progenitors ( BFU and CFU ) and primitive cells ( CAFC , LTC-IC ) were analyzed by methylcellulose assay . Phenotypic evolution and clonogenic properties of the lin(-) population were analyzed along erythroid and megakaryocytic differentiation . RESULTS : We observed P38936 ectopic expression in early hematopoietic progenitors ( lin(-)Sca(+) ) , megakaryocytes , and , to a lesser extent , erythroid cells . This expression induced an important decrease in the number of CFU-MK , BFU-E , CFU-E , primitive progenitors ( CAFC day 35 ) , and LTC-IC , but did not affect the maturation process of these cells and the blood cell count . CONCLUSIONS : We show that variation of P38936 expression level changes the fate of hematopoietic cells by favoring either proliferation or differentiation pathways . This effect of P38936 is exerted not only at the level of primitive progenitors but also in more mature progenitors . However , in vivo , a systemic compensation mechanism is most likely activated in response to variations of the flow of progenitor production . New antithrombotics for the treatment of acute and chronic arterial ischemia . The established antithrombotic agents are effective but they have limitations which have provided opportunities for the development of new antithrombotic compounds . Of these new agents , the antithrombin III-independent thrombin inhibitors and the platelet P08514 /IIIa receptor antagonists are the most advanced in their development . Other new antithrombotic agents include the antithrombin III-independent factor Xa inhibitors , activated protein C , soluble thrombomodulin and tissue factor pathway inhibitor . Of the P08514 /IIIa antagonists , the humanized DB00054 and integrin have been evaluated in phase III studies . The DB00054 was effective in preventing both short-term and longer-term complications of coronary angioplasty . The antithrombin III-independent thrombin inhibitors hirudin and hirulog have also been evaluated in phase III studies . The studies with hirudin as an adjuvant to coronary thrombolysis had to be terminated and restarted at lower dosages because of an unacceptable incidence at intracranial hemorrhage and the study with hirulog produced equivocal results . Genetic polymorphisms for the study of multifactorial stroke . Single-gene disorders explain only a minority of stroke cases . Stroke represents a complex trait , which is usually assumed to be polygenic . On this topic , the role of a wide number of candidate genes has been investigated in stroke through association studies , with controversial results . Therefore , it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke . This review is an update and an extensive analysis of the more recent association studies conducted in stroke . We evaluated a number of studies on several candidate genes ( including P12259 , F2 , P02671 / P02675 / P02679 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / Q9UKI9 / P04054 / P08514 , P17301 , P07359 , P12821 , AGT , NOS3 , P02649 , P06858 , P27169 , Q08499 , P20292 , P42898 , Q99707 , and P35520 ) , providing a final panel of genes and molecular variants . We categorized this panel in relation to the degree of association with stroke , supported by the results of meta-analyses and case-control studies . Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses . DB01109 potentiation of collagen-induced platelet aggregation is related to the P08514 / P05106 receptor and not to the GPIb receptor , as tested by whole blood aggregometry . To determine whether heparin potentiation of platelet aggregation is related to platelet GP IIb/IIIa and GP Ib receptors , four series of experiments were performed on blood from normal volunteers . In the first experiment pretreatment with the monoclonal antibody DB00054 ( MAb DB00054 ) , which antagonizes at the GP IIb/IIIa receptor , potently inhibited the collagen-induced platelet aggregation ( p less than 0.001 ) . With heparin added to blood pretreated with MAb DB00054 , the aggregation increased ( p less than 0.005 ) to an extent similar to that when only saline was used for pretreatment . In the second experiment , monoclonal antibody 10E5 ( MAb 10E5 ) and peptide RGDS , substances which also antagonize at the GP IIb/IIIa receptor , decreased collagen-induced platelet aggregation to an extent similar to that after pretreatment with MAb DB00054 . Following pretreatment with RGDS , heparin increased platelet aggregation ( p less than 0.03 ) , while after pretreatment with antibody MAb 10E5 heparin did not enhance platelet aggregation . In the third experiment aurin , an inhibitor of P04275 and its interaction with the platelet GPIb receptor , decreased platelet aggregation dose-dependently . In the fourth experiment heparin enhanced platelet aggregation to a similar extent ( p less than 0.005 ) , regardless of pretreatment of the blood with saline , aurin or monoclonal antibody 6D1 ( MAb 6D1 ) , the latter an antagonist at the GP Ib receptor . In conclusion , the potentiation of collagen-induced platelet aggregation by heparin was not inhibited by MAb DB00054 , RGDS , aurin or MAb 6D1 , but was abolished by MAb 10E5 , implying that the heparin effect is related to activation of the platelet GP IIb/IIIa receptor complex . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Inhibition of poly(ADP-ribose) polymerase enhances cell death and improves tumor growth delay in irradiated lung cancer models . PURPOSE : Poly(ADP-ribose) polymerase-1 ( P09874 ) is the founding member of a family of enzymes that catalyze the addition of ADP-ribose units to proteins that mediate DNA repair pathways . Ionizing radiation induces DNA strand breaks , suggesting that P09874 inhibition may sensitize tumor cells to radiation . EXPERIMENTAL DESIGN : We investigated the combination of P09874 inhibition with radiation in lung cancer models . ABT-888 , a novel potent P09874 inhibitor , was used to explore the effects of P09874 inhibition on irradiated tumors and tumor vasculature . RESULTS : ABT-888 reduced clonogenic survival in H460 lung cancer cells , and inhibited DNA repair as shown by enhanced expression of DNA strand break marker histone gamma- P16104 . Both apoptosis and autophagy contributed to the mechanism of increased cell death . Additionally , ABT-888 increased tumor growth delay at well-tolerated doses in murine models . For a 5-fold increase in tumor volume , tumor growth delay was 1 day for ABT-888 alone , 7 days for radiation alone , and 13.5 days for combination treatment . Immunohistochemical staining of tumor sections revealed an increase in terminal deoxyribonucleotide transferase-mediated nick-end labeling apoptotic staining , and a decrease in Ki-67 proliferative staining after combination treatment . Matrigel assay showed a decrease in in vitro endothelial tubule formation with ABT-888/radiation combination treatment , and P04275 staining of tumor sections revealed decreased vessel formation in vivo , suggesting that this strategy may also target tumor angiogenesis . CONCLUSIONS : We conclude that P09874 inhibition shows promise as an effective means of enhancing tumor sensitivity to radiation , and future clinical studies are needed to determine the potential of ABT-888 as a radiation enhancer . The cytokines ( P01579 , P60568 , P05112 , P22301 , Q16552 ) and Treg cytokine ( TGF-beta1 ) levels in adults with immune thrombocytopenia . Previous studies have indicated that autoimmune diseases might be caused by an imbalance of T helper cells ( Th ) , cytokines , and regulatory T cells ( Treg ) cytokines . We measured the plasma concentrations of Th1-associated cytokines ( P01579 , P60568 ) , Th2 -associated cytokines ( P05112 , P22301 ) , Th17-associated cytokine ( Q16552 ) and Treg -associated cytokine ( TGF-beta1 ) in adult patients with immune thrombocytopenia ( ITP ) and evaluated their clinical relevance . Plasma P01579 , P60568 , P05112 , P22301 , Q16552 and TGF-beta1 concentrations of 52 ITP patients and 30 age- and sex-matched healthy controls were measured by enzyme-linked immunosorbent assay method ( ELISA ) . Concentration of Th2 cytokines ( P05112 and P22301 ) were significantly higher in ITP patients compared to controls ( P < 0.05 ) . However , concentrations of Th1 cytokines ( P01579 , P60568 ) , Th17 cytokine ( Q16552 ) and Treg cytokine ( TGF-beta1 ) were lower in ITP patients ( P < 0.05 ) . Concentration of Q16552 was significantly higher in chronic ITP patients compared to severe ITP patients ( P < 0.05 ) , and no significant difference of cytokine concentration among the other subgroups in ITP patients was found . Among the ITP patients , concentration of P01579 correlated positively and significantly with PAIgG ( r = 0.48 , P = 0.02 ) . A significant correlation was neither found between other cytokine levels and platelet count , nor between cytokine levels and megakaryocytes number , nor between cytokines levels and PAIgG or P08514 /IIIa and/or GPIb/IX autoantibodies . The present study demonstrates that an imbalance of Th and Treg cytokines may mediate the pathogenesis of ITP . 7E3 F(ab')2 , an effective antagonist of rat alphaIIbbeta3 and alphavbeta3 , blocks in vivo thrombus formation and in vitro angiogenesis . DB00054 ( c7E3 Fab , ReoPro ) blocks P08514 /IIIa and alphavbeta3 and inhibits thrombotic and proliferative events only in humans and non-human primates . The bivalent F(ab')2 fragment is an effective anti-thrombotic agent in canine models . In the present study , 7E3 F(ab')2 was also found to bind to rat P08514 /IIIa ( KD = 27 +/- 4 microg/mL ) and alphavbeta3 ( KD = 9 +/- 8 microg/mL ) , to block in vitro rat platelet aggregation ( IC50 = 16 +/- 6 microg/mL ) , and to inhibit alphavbeta3-mediated microvessel sprout formation in a rat aortic ring assay . Following administration of 7E3 F(ab')2 ( 4 mg/kg ) to rats , platelet aggregation was completely blocked for up to 6 h and thrombus formation in response to a rat abdominal aorta double crush injury was prevented . Effective chronic dosing was achieved with 6 mg/kg daily I.P. injections . In vitro mixing experiments indicated that 7E3 F(ab')2 redistributed to unlabeled platelets in 2 h . Ex vivo , 7E3 F(ab')2 was detected on platelets for up to 4 days after a single 4-mg/kg injection . These data suggest that 7E3 F(ab')2 may be a useful agent to study the effects of P08514 /IIIa and alphavbeta3 blockade in rat models of thrombosis and vascular disease . The high molecular mass , glycoprotein Ib-binding protein flavocetin-A induces only small platelet aggregates in vitro . The direct effects of snake venom glycoprotein ( GP ) Ib-binding proteins on platelet receptors during the formation of platelet aggregates were determined by a particle counting method using light scattering . Flavocetin-A induces small platelet aggregates , but not medium or large ones . However , neither jararaca GPIb-BP nor tokaracetin induce platelet aggregation . The flavocetin-A dose-response curve for formation of small aggregates is bell-shaped , with maximal effect at 1 to 2 microg/mL . The formation of small aggregates was not observed when fixed human platelets were used . Jararaca GPIb-BP , the anti-GPIb monoclonal antibody GUR83-35 , prostaglandin I2 , and ethylene diamine-N,N-dimethylformamide all inhibited flavocetin-A-induced small aggregate formation , but acetylsalicylic acid did not . Furthermore , anti- P08514 /IIIa monoclonal antibodies , DB00054 , and YM337 significantly but partially inhibited aggregate formation , but the anti- P04275 monoclonal antibody NMC-4 had no effect . The formation of small aggregates required extracellular calcium , but flavocetin-A did not elevate cytosolic calcium . These results suggest that flavocetin-A binds to intact platelets , initiating platelet responses and inducing platelet aggregate formation by cross-linking platelets . Consequently , flavocetin-A may be a useful tool to study the mechanism of GPIb-mediated platelet activation and the structure-function relationships of GPIb . No evidence for an influence of the human platelet antigen-1 polymorphism on the antiplatelet effects of glycoprotein IIb/IIIa inhibitors . This study investigated the hypothesis that the human platelet antigen-1 ( Q9Y251 -1 ) polymorphism may influence the antiplatelet effects of glycoprotein (GP)IIb/IIIa inhibitors . DB00640 diphosphate ( 30 micro mol ) -induced fibrinogen binding was measured by flow cytometry . DB00054 ( 0.03-3 micro g/ml ) , tirofiban ( 0.3-30 nmol/l ) or eptifibatide ( 0.01-1 micro g/ml ) were incubated for 15 min with the samples prior to stimulation . IC(50) values for the inhibition of fibrinogen binding were determined from each experiment . All subjects were genotyped by GALIOS and automated fluorescence correlation spectroscopy . Although a marked variability in the inhibitory effects of all three P08514 /IIIa inhibitors was confirmed , there were no significant differences between the genotypes with respect to the inhibition of fibrinogen binding . Thus , the present study does not provide evidence for an effect of Q9Y251 -1 polymorphism on the inter-individual variability in the platelet inhibitory effects of the three P08514 /IIIa inhibitors approved for clinical use . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . 7E3 F(ab')2 , a monoclonal antibody to the platelet P08514 /IIIa receptor , protects against microangiopathic hemolytic anemia and microvascular thrombotic renal failure in baboons treated with C4b binding protein and a sublethal infusion of Escherichia coli . We have used our previously described baboon model of infusion of both a sublethal dose of Escherichia coli and C4b binding protein to assess the impact of inhibiting platelet function with the F(ab')2 fragment of the monoclonal antibody DB00054 , directed against the platelet glycoprotein (GP)IIb/IIIa receptor , on the characteristic microvascular changes . At a dose of 0.25 to 0.35 mg/kg bolus plus an infusion of 0.25 to 0.35 mg/kg over 6 hours , c7E3 F(ab')2 had only a minimal impact on fibrinogen consumption and delayed but did not prevent , the development of thrombocytopenia . Treatment with 7E3 F(ab')2 , however , produced significant protection from the development of microangiopathic hemolysis and renal insufficiency . Histologic examination supported these observations , with treated animals having fewer schistocytes on blood smear and less evidence of ischemic renal changes . Treated animals also had more rapid recovery of peripheral white blood counts , suggesting a possible protective effect of treatment on ischemic damage to the bone marrow . These data indicate that potent inhibition of platelet function via P08514 /IIIa receptor blockade can decrease ischemic organ damage in this animal model that has features similar to those found in diffuse intravascular coagulation , hemolytic uremic syndrome , and thrombotic thrombocytopenic purpura . Anti-thrombotic and anticoagulant treatment in interventional cardiology . Efforts to improve Percutaneous Transluminal Coronary Angioplasty ( PTCA ) have resulted in the usage of new antiplatelets , and antithrombotic agents . These new agents may increase bleeding complications . However , EPIC , EPILOG and CAPTURE trials showed benefits of DB00054 , a P08514 /IIIa platelets receptor blocker , in high risk PTCA patients . On the other hand , direct thrombin inhibitors , Hirudin and DB02351 , did not clearly show any benefit when compared to heparin in patients with unstable angina undergoing PTCA . Combination of oral antiplatelets , ticlopidine and aspirin , is widely utilized following stent implantation . However , its benefit over aspirin alone has not been demonstrated . This article aims to review mechanisms and benefits of these new agents in cardiovascular field . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Conjugation and evaluation of 7E3 x P4B6 , a chemically cross-linked bispecific F(ab')2 antibody which inhibits platelet aggregation and localizes tissue plasminogen activator to the platelet surface . A bispecific F(ab')2 monoclonal antibody which recognizes both the platelet P08514 /IIIa receptor and human tissue plasminogen activator was produced to target tPA to platelets for enhancement of thrombolysis . A stable , thioether-cross-linked bispecific F(ab')2 ( 7E3 X P4B6 ) combining the P08514 /IIIa-specific monoclonal antibody DB00054 , which inhibits platelet aggregation , and a nonneutralizing anti-tPA monoclonal antibody ( P4B6 ) was produced . This was performed by coupling each of the parental Fab ' moieties with the homobifunctional cross-linker bis ( maleimido methyl ) ether ( BMME ) . 7E3 X P4B6 was sequentially purified using gel-filtration chromatography and hydrophobic interaction ( HIC ) HPLC . HIC was shown to completely resolve each of the parental F(ab')2 species from the bispecific one . 7E3 X P4B6 was shown to retain completely each of the parental immunoreactivities in P08514 /IIIa and tPA binding EIA 's . The bispecific antibody inhibited platelet aggregation in vitro at levels comparable to those for 7E3 Fab . Recruitment of tPA activity to washed human platelets was demonstrated using the S-2251 chromogenic substrate assay . 7E3 X P4B6 recruited 12-fold more tPA to the washed platelets than a mixture of the parental F(ab')2 molecules used as controls . Q13444 is an adhesion receptor for platelet P08514 -IIIa and induces platelet activation . Cell adhesion and proteolytic matrix degradation are central processes in atherosclerosis . Being a member of the family of ADAMs ( " a disintegrin and metalloproteinase " ) , metargidin ( Q13444 ) combines a metalloproteinase domain and an RGD aminoacid sequence . We studied the potential role of Q13444 as an adhesion receptor on endothelial cells and interactions between platelets and Q13444 with respect to platelet adhesion , activation and thrombus formation . Q13444 was found to be expressed on cultured endothelial cells ( HUVEC ) . Platelet adhesion to immobilized recombinant Q13444 was effectively enhanced under both static and high shear rate conditions reaching the maximum level of adhesion to fibrinogen . Consistently , platelet adhesion onto Q13444 overexpressing endothelial cells was significantly increased . Adhesion to Q13444 was reduced by blockade of P08514 -IIIa using neutralizing anti-alpha(IIb)beta3 mAbs ( DB00054 , 2G12 ) , but not by anti-alpha(v)beta3 ( LM609 ) . Soluble Q13444 binds to activated but not to resting P08514 -IIIa . Moreover , platelets adherent to Q13444 additionally attracted platelets under high shear rates indicating an initial role of platelet- Q13444 interactions for thrombus formation . Furthermore , incubation of platelets with soluble Q13444 showed a dose-dependent increase in secretion of CD62P and P29965 . Q13444 is expressed on endothelial cells and can serve as an adhesion receptor for platelets via P08514 -IIIa binding . Platelet adhesion to Q13444 leads to platelet activation , secretion and promotes thrombus formation . Thus , Q13444 may represent a novel target for antithrombotic strategies in cardiovascular pathologies . The effect of s-nitroso-glutathione on platelet and leukocyte function during experimental extracorporeal circulation . Treatment with extracorporeal membrane oxygenation ECMO ) is associated with side effects , e.g. , blood cell consumption and activation . Our group has earlier shown that nitric oxide administered as a gas reduces platelet consumption and activation . In the present work we have studied the effect of the NO-donor S-nitroso-glutathione GSNO ) on platelets and leukocytes in an in vitro extracorporeal circuit . Two complete ECMO circuits were perfused with fresh heparinized human blood for 24 hours . GSNO was administered as a continuous infusion to one circuit at a rate of 0.7 mg/hour in four paired experiments and at a rate of 3.5 mg/hour in another four paired experiments . The other circuit was used as a control . Blood samples were withdrawn from both circuits before the start of the experiments and at 0.5 , 1 , 3 , 12 , and 24 hours of perfusion . The samples were analyzed for red blood cell count , leukocyte count , platelet count , platelet membrane expression of glycoproteins GP ) Ib and P08514 /IIIa , leukocyte membrane expression of cluster of differentiation CD ) 11b/ P05107 , as well as plasma concentration of tumor necrosis factor P01375 ) -alpha , interleukin IL ) -1beta , and P10145 . No difference in these parameters between the GSNO and the control circuit at any time point was assayed . In this study , no significant effect of GSNO on circulating platelets or leukocytes during experimental extracorporeal circulation could be shown . Antiaggregatory and proangiogenic effects of a novel recombinant human dual specificity anti-integrin antibody . BACKGROUND : beta(3)-Integrins are involved in platelet aggregation via alpha(IIb)beta(3) [ glycoprotein (GP)IIb- P05106 ] , and in angiogenesis via endothelial alpha(V)beta(3) . Cross-reactive ligands with antiaggregatory and proangiogenic effects , both desirable in peripheral vasculopathies , have not yet been described . OBJECTIVES : In vitro and in vivo characterization of antiaggregatory and proangiogenic effects of two recombinant human Fab fragments , with emphasis on beta(3)-integrins . METHODS : Recombinant Fab fragments were obtained by phage display technology . Specificity , affinity and IC(50) were determined by immunodot assays , enzyme-linked immunosorbent assay ( ELISA ) , and Scatchard plot analysis , and by means of human umbilical vein endothelial cells ( HUVECs ) . Functional analyses included ELISA for interaction with fibrinogen binding to P08514 - P05106 , flow cytometry for measurement of activation parameters and competitive inhibition experiments , human platelet aggregometry , and proliferation , tube formation and the chorioallantoic membrane ( P62158 ) assay for measurement of angiogenic effects . RESULTS : We observed specific and high-affinity binding to an intact P08514 - P05106 receptor complex of two human Fab autoantibody fragments , with no platelet activation . Dose-dependent fibrinogen binding to P08514 - P05106 and platelet aggregation were completely inhibited . One Fab fragment was competitively inhibited by abciximab and its murine analog monoclonal antibody ( mAb ) DB00054 , whereas the other Fab fragment bound to cultured HUVECs , suggesting cross-reactivity with alpha(V)beta(3) , and also demonstrated proangiogenic effects in tube formation and P62158 assays . CONCLUSIONS : These Fab fragments are the first entirely human anti- P08514 - P05106 Fab fragments with full antiaggregatory properties ; furthermore , they do not activate platelets . The unique dual-specificity anti-beta(3)-integrin Fab fragment may represent a new tool for the study and management of peripheral arterial vasculopathies . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . Repetitive profound thrombocytopenia after treatment with tirofiban : a case report . The P08514 /IIIa inhibitors are used in the acute coronary syndromes and interventional cardiology as antiplatelet agents . These drugs induce thrombocytopenia in approximately 1-5 % of patients . Thrombocytopenia is rapid in onset and antibody mediated . DB00054 is associated with higher incidence of thrombocytopenia than eptifibatide and tirofiban . Profound thrombocytopenia has reportedly been an issue with abciximab , but not with tirofiban . We reported a case of acute profound thrombocytopenia due to on tirofiban treatment in the same patient at two different times . Increased thromboxane biosynthesis during coronary thrombolysis . Evidence that platelet activation and thromboxane A2 modulate the response to tissue-type plasminogen activator in vivo . Platelet activation is markedly increased during coronary thrombolysis and limits the response to thrombolytic therapy . A possible mediator of platelet activation in this setting is thromboxane ( TX ) A2 , a potent platelet agonist formed in greatly increased amounts during coronary thrombolysis in man . To address this hypothesis , we examined the role of TXA2 in modulating the response to intravenous tissue-type plasminogen activator ( t-PA ) in a chronic canine model of coronary thrombosis . Reperfusion occurred in 60 +/- 5 minutes and was complicated by spontaneous reocclusion . The times to reperfusion and reocclusion were platelet-dependent . Consistent with a role for TXA2 in this process , TXA2 biosynthesis , determined a excretion of its enzymatic metabolite , 2,3-dinor-TXB2 , was markedly increased during coronary thrombolysis . Furthermore , inhibition of TXA2 by aspirin , given alone or in combination with a TXA2/prostaglandin endoperoxide receptor antagonist , accelerated reperfusion and partly inhibited cyclic flow variations during reperfusion . The delay in reperfusion and reocclusion induced by TXA2 appeared to be mediated by platelet aggregation since the F(ab')2 fragment of DB00054 , a monoclonal antibody to the platelet P08514 /IIIa , also accelerated reperfusion and prevented reocclusion without altering TXA2 biosynthesis . These finding suggest that platelet aggregation limits the response to coronary thrombolysis and that platelet activation in this setting is partly TXA2-dependent . Potential future clinical applications for the P08514 /IIIa antagonist , abciximab in thrombosis , vascular and oncological indications . DB00054 ( ReoPro ) is a mouse-human chimeric monoclonal antibody Fab fragment of the parent murine monoclonal antibody DB00054 , and was the first of these agents approved for use as adjunct therapy for the prevention of cardiac ischemic complications in patients undergoing percutaneous coronary intervention ( P05154 ) . DB00054 binds with high avidity to both the non-activated and activated form of the P08514 /IIIa receptor of platelets , the major adhesion receptor involved in aggregation . Additional cardiovascular indications for abciximab are unstable angina , carotid stenting , ischemic stroke and peripheral vascular diseases . DB00054 also interacts with two other integrin receptors ; the a av b b3 receptor , which is present in low numbers on platelets but in high density on activated endothelial and smooth muscle cells , and a aMb b2 integrin which is present on activated leukocytes . Cell types that express integrins P08514 /IIIa and a av b b3 such as platelets , endothelial and tumor cells have been implicated in angiogenesis , tumor growth and metastasis . Since abciximab interacts with high avidity to integrins P08514 /IIIa and a av b b3 , it is reasonable to assume that it may possess anti-angiogenic properties in angiogenesis-related diseases , as well as anti-metastastatic properties in case of disseminating tumors expressing the target integrin receptors . Spa bathing activates fibrinolysis in patients with cerebral infarction . The effects of spa bathing on blood coagulation and fibrinolysis were studied in 20 patients with chronic cerebral infarction . Blood was obtained before and after a 10-minute period of spa bathing at 41 degrees C . P00734 time , activated partial thromboplastin time , fibrinogen , factor VIII activity , P04275 activity , and antithrombin III activity did not show significant changes after bathing , but euglobulin lysis time was significantly reduced ( p < 0.01 ) and fibrin lysis activity was increased ( p < 0.05 ) . These findings suggest that spa bathing activates fibrinolysis without markedly changing blood coagulation in patients with chronic cerebral infarction . It is thought that the activation of fibrinolysis without the activation of coagulation has a favorable effect on blood circulation . The results of fibrin-plate assays using C1 inactivator indicated that tissue-type plasminogen activator was the major contributor to the activation of fibrinolysis during spa bathing . MHC class I-associated peptides identified from normal platelets and from individuals with idiopathic thrombocytopenic purpura . Major histocompatibility complex ( MHC ) class I molecules bind and display peptide antigens on the cell surface . CD8(+) T lymphocytes recognize peptides in association with class I proteins to initiate a cytotoxic immune response . To understand the specificity of such immune responses and to facilitate the development of therapies for disease , it is important to identify MHC-presented peptides . In this study , platelets , easily obtainable and often associated with immune-mediated disease , were selected to identify MHC class I-associated peptides . MHC-associated peptides presented on platelets of normal individuals and individuals with idiopathic thrombocytopenic purpura ( ITP ) were characterized . ITP is characterized by the premature immune destruction of platelets . It is associated with the production of antiplatelet autoantibodies , most often targeting platelet membrane P08514 /IIIa or GPIb/IX . In addition to characterizing five fully and several partially sequenced peptides from platelets , the peptide GPRGA(L/I)S(L/I)(L/I) was identified from four of the five ITP patients . The anchor motif of this peptide correlates with the presence of the HLA- P33681 allele . A BLAST search identified this peptide as GPIb ( 4-12 ) . In conclusion , platelets from normal and ITP individuals can present peptides from general cellular proteins and platelet specific proteins , such as GPIb , to the immune system via MHC class I . Monoclonal antibody against the platelet glycoprotein ( GP ) IIb/IIIa receptor prevents coronary artery reocclusion after reperfusion with recombinant tissue-type plasminogen activator in dogs . Localized thrombosis was produced in the left anterior descending ( LAD ) coronary artery of open chest dogs by constricting a segment so as to produce greater than 90 % stenosis ( reducing blood flow to 40 +/- 10 % of baseline ) , and placing a thrombus in the segment immediately proximal to the stenosis by inducing endothelial cell injury and instilling a mixture of blood and thrombin . Intravenous infusion of recombinant tissue-type plasminogen activator ( rt-PA ) at a rate of 15-30 micrograms/kg per min for 30 or 60 min in eight dogs induced coronary artery reperfusion within 23 +/- 7 min ( mean +/- SD ) , but reocclusion occurred despite heparin anticoagulation in all but one of these dogs within 7 +/- 5 min . Intravenous injection of 0.8 mg/kg of the F(ab')2 fragment of a monoclonal antibody ( DB00054 ) directed against the platelet P08514 /IIIa receptor , prevented reocclusion in 10/10 dogs during an observation period of 2 h ( P less than 0.001 vs. rt-PA alone ) . The antibody abolished ADP-induced platelet aggregation and markedly prolonged the bleeding time . Intravenous aspirin or dipyridamole prevented reocclusion for 1 h or more in only 2/7 and 1/6 dogs , respectively . We conclude that the monoclonal antibody is very effective in preventing reocclusion after successful thrombolysis of occluded coronary arteries with rt-PA . A review of the mechanisms and effectiveness of dietary polyphenols in reducing oxidative stress and thrombotic risk . Dietary sources of polyphenols , which are derivatives and/or isomers of flavones , isoflavones , flavonols , catechins and phenolic acids , possess antioxidant properties and therefore might be important in preventing oxidative-stress-induced platelet activation and attenuating adverse haemostatic function . Free radicals , including reactive oxygen and nitrogen species , promote oxidative stress , leading to platelet hyperactivation and the risk of thrombosis . The consumption of antioxidant/polyphenol rich foods might therefore impart anti-thrombotic and cardiovascular protective effects via their inhibition of platelet hyperactivation or aggregation . Most commonly-used anti-platelet drugs such as aspirin block the cyclooxygenase ( P36551 ) -1 pathway of platelet activation , similar to the action of antioxidants with respect to neutralising hydrogen peroxide ( H2 O2 ) , with a similar effect on thromboxane production via the P23219 pathway . Polyphenols also target various additional platelet activation pathways ( e.g. by blocking platelet-ADP , collagen receptors ) ; thus alleviating fibrinogen binding to platelet surface ( P08514 -IIIa ) receptors , reducing further platelet recruitment for aggregation and inhibiting platelet degranulation . As a result of the ability of polyphenols to target additional pathways of platelet activation , they may have the potential to substitute or complement currently used anti-platelet drugs in sedentary , obese , pre-diabetic or diabetic populations who can be resistant or sensitive to pharmacological anti-platelet therapy . DB00054 as an adjunctive therapy for patients undergoing percutaneous coronary interventions . INTRODUCTION : Platelets play a central role in the pathophysiology of acute coronary syndromes ( ACS ) and activation of platelet glycoprotein ( GP ) IIb/IIIa receptor is critical to platelet aggregation . DB00054 , a human murine chimeric antibody to the P08514 /IIIa receptor , is an important biological therapy in the management of patients presenting with ACS . AREAS COVERED : The objective of this review is to define the role of abciximab in the management of ACS by interpreting the available data from randomized clinical trials using abciximab in various clinical scenarios , particularly in percutaneous coronary intervention ( P05154 ) . We also review different modes of delivery and describe the adverse effects of abciximab including thrombocytopenia . Where possible , we attempt to compare abciximab to the other available P08514 /IIIa inhibitors . We hope the reader will gain a better understanding of the benefits and risks of abciximab and the important role it has in the management of cardiology patients . EXPERT OPINION : DB00054 was a breakthrough drug in the management of high risk ACS patients undergoing P05154 . However , with newer available therapies and improvement in P05154 technology , dose and delivery of this drug have evolved as we try to extract maximum benefit while minimizing the adverse effects associated with it . Activating peroxisome proliferator-activated receptor gamma mutant promotes tumor growth in vivo by enhancing angiogenesis . P37231 ( PPARgamma ) is expressed in a variety of cancer cells . The addition of ligand activates the receptor by inducing a conformational change in the receptor , which can be recapitulated by mutation . To investigate the role of activated PPARgamma signaling in breast cancer , we compared the function of a constitutively active PPARgamma ( PgammaCA ) mutant with the wild-type PPARgamma in ErbB2-induced mammary tumorigenesis in vivo . Tumor cells transduced with either PPARgamma or PgammaCA were implanted into immunocompetent FVB mice . Enhanced tumor growth was observed in PgammaCA-transduced cells , which was associated with increased angiogenesis and endothelial stem cells as evidenced by increased number of cells stained with P04275 , c-Kit , CD133 , and CD31 . Genome-wide expression profiling identified a group of genes within the angiogenesis pathway , including Angptl4 , as targets of activated PPARgamma ; PgammaCA also induced Angptl4 protein secretion in ErbB2-transformed mammary epithelial cells . Angptl4 promoted vascular endothelial cell migration ; conversely , immunodepletion of Angptl4 reduced PgammaCA-mediated cellular migration . Collectively , these studies suggest that activated PPARgamma induces Angptl4 to promote tumor growth through enhanced angiogenesis in vivo . Prevention of rethrombosis after coronary thrombolysis in a chronic canine model . I . Adjunctive therapy with monoclonal antibody 7E3 F(ab')2 fragment . We examined the efficacy of the monoclonal antibody ( MoAb ) 7E3 F(ab')2 fragment , an inhibitor of the platelet glycoprotein (GP)IIb/IIIa receptor , to prevent coronary artery rethrombosis after successful thrombolysis with rt-PA . The circumflex coronary artery of anesthetized dogs was instrumented with a flow probe , an electrode , and a stenosis . After recovery from the surgical procedure , the animals were reanesthetized on post-operative day 9 , and vessel wall injury was induced with current applied to the intimal surface of the circumflex coronary artery . The resulting occlusive thrombus was aged for 30 min , and recombinant tissue plasminogen activator ( rt-PA ) was administered . The animals were allocated to receive either placebo or a single dose of DB00054 [ 0.8 mg/kg intravenous ( i.v. ) bolus ] as the sole adjunctive agent . Ex vivo platelet function and coronary artery blood flow velocity were recorded on each of 5 consecutive days . Reocclusion and mortality were reduced significantly in animals treated with 7E3 as compared with the placebo-treated group . Significant inhibition of ex vivo platelet aggregation persisted for 48 h after a single injection of DB00054 . The MoAb 7E3 F(ab')2 fragment is effective as the sole adjunctive agent with rt-PA for prevention of rethrombosis . The present study is unique in that it examined the efficacy of P08514 /IIIa inhibition in an experimental model for an extended time , demonstrating the duration of antiplatelet therapy required to prevent rethrombosis after thrombolysis . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . P08514 /IIIa antagonists and other anti-integrins . Platelet aggregation involves the binding of adhesive proteins ( fibrinogen , P04275 ) to the alphaIIbbeta3 integrin , which assumes a high-affinity state for adhesive proteins during platelet activation . The occupied integrin sends signals back into the platelet , and the bound adhesive protein forms the bridges linking platelets together . Anti-integrin therapy is designed to inhibit this process in arterial thrombosis . DB00054 , mouse-human chimeric Fab fragments , blocks platelet aggregation and provides proven clinical benefit in acute situations such as in patients with unstable angina undergoing angioplasty or stenting . DB00063 and tirofiban are small molecular mass inhibitors also in current use . In contrast , oral inhibitors of alphaIIbbeta3 have proved disappointing , provoking increased mortality without assuring an adequate blockade of alphaIIbbeta3 . The problems of using anti-integrin therapy are discussed in this article as are ways of improving its efficacity . Final thoughts provide ideas for a new generation of inhibitors . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . New anticoagulant strategies . The limitations of standard heparin have prompted the development of a variety of newer antithrombotic agents . In fact , a LMWH preparation has recently been approved for clinical use in North America . Of these novel preparations , LMWH , the direct thrombin inhibitors , and inhibitors of P08514 -IIIa have been used clinically and are in advanced stages of evaluation . Not only is LMWH effective in the prevention of venous thromboembolic disease in high-risk patients , but its more predictable dose response makes it an ideal candidate for the treatment of venous thrombosis . Further studies are needed to determine whether LMWH is superior to standard heparin as adjunctive therapy in patients undergoing coronary thrombolysis or angioplasty . Particularly promising in the setting of arterial thrombosis are hirudin , hirulog , and DB00054 . With the encouraging results reported to date , it is likely that these agents will soon find their way into the treatment armamentarium of arterial thrombosis . Analysis of P08514 /IIIa receptor number by quantification of 7E3 binding to human platelets . A large number of glycoprotein ( GP ) IIb/IIIa receptors are present on the surface of platelets . Studies to define precisely the number of P08514 /IIIa receptors using specific monoclonal antibodies ( MoAbs ) or fibrinogen binding have , however , yielded varying estimates of receptor number . To refine the quantitative estimation of P08514 /IIIa receptors on resting platelets , we have used the MoAb DB00054 , which has high affinity for P08514 /IIIa . Quantitative binding studies were performed using radiolabeled conjugates of 7E3 IgG , as well as fragments and derivatives of DB00054 . For platelets obtained from any single individual , the numbers of 7E3 F(ab')2 and IgG molecules bound per platelet were equivalent ( approximately 40,000 ) , whereas the number of Fab molecules bound per platelet was consistently approximately twofold higher ( approximately 80,000 ) . To investigate the basis of the quantitative disparity in binding of intact 7E3 and 7E3 F(ab')2 versus 7E3 Fab , we studied the binding of a newly constructed , bispecific (Fab')2 molecule containing only a single 7E3 combining site . Because this construct bound to the same extent as the Fab species , the larger size of the intact 7E3 and 7E3 F(ab')2 molecules could not explain the reduced number of molecules that bound per platelet compared to the Fab fragment . Rather , it appears that the valency of the antibody is the critical factor determining the number of antibody molecules bound per platelet . Thus , we conclude that the binding of 7E3 Fab corresponds most closely with surface P08514 /IIIa number and that the number of P08514 /IIIa receptors is approximately 80,000 per platelet . Arterial reocclusion and persistent distal occlusion after thrombus aspiration . BACKGROUND AND PURPOSE : Early reocclusion of intracranial arteries can lead to poor clinical outcome . We report reocclusion detection after endovascular clot aspiration , followed by administration of P08514 -IIIa antagonist under continuous ultrasound monitoring . CASE DESCRIPTION : A 73-year-old man developed the right middle cerebral artery ( MCA ) occlusion with NIHSS 17 points , 6 days after aortic valve replacement . Recanalization was achieved with Penumbra™ system and reocclusion was detected with transcranial Doppler ( P24386 ) 30 minutes postcompletion of intra-arterial procedure . Proximal recanalization was achieved with the second thrombus aspiration while M2 MCA occlusion persisted beyond the reach of the device . Intravenous abciximab was administered under continuous P24386 monitoring . Recanalization with Thrombolysis in Brain Ischemia ( TIBI ) flow grade 4 was observed at 60 minutes postintervention accompanied with clinical recovery to NIHSS 3 points . DB00054 was given for 12 hours with no hemorrhagic transformation on repeat CT scan . Patient was discharged home with mild left pronator drift and facial droop , and his modified ranking score was 1 at 6-week follow-up visit . CONCLUSIONS : Early arterial reocclusion can occur after successful thrombus aspiration while P08514 -IIIa antagonist administration may lead to subsequent recanalization of persisting distal occlusions not amenable to mechanical removal . Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation . The platelet integrin glycoprotein ( GP ) IIb/IIIa , which mediates platelet aggregation , has been the target for novel antiplatelet agents , the P08514 /IIIa antagonists . Several P08514 /IIIa antagonists have been developed based on the peptide RGDS present in adhesion proteins , including the principle ligand fibrinogen . The apoptosis enzyme , procaspase-3 , contains an RGD-recognition sequence and is activated by RGDS . We examined the effects of RGDS and several P08514 /IIIa antagonists on cell death and procaspase-3 activation in rat neonatal cardiomyocytes . These antagonists do not recognize rat integrins , yet RGDS , orbofiban , and xemilofiban induced dose-dependent apoptosis and procaspase-3 activation in cardiomyocytes over 72 h , particularly under hypoxic conditions . Scrambled peptide , the monoclonal antibody 7E3 or integrelin ( a peptide containing a KGD sequence ) , had little or no effect . Immunoprecipitation of procaspase-3 followed by treatment with the compounds showed that procaspase-3 was activated directly by RGDS , orbofiban , xemilofiban , and by monoclonal DB00054 , the latter demonstrating that compounds must enter cells to induce apoptosis through caspase activation . DB00063 had no effect . Binding studies with (3)H-SC52012B , a P08514 /IIIa antagonist analogue of orbofiban , showed no specific binding to cardiomyocytes , but the radioligand accumulated intracellularly over 72 h . (3)H-SC52012B also bound directly to human recombinant caspase-3 ( K(d) , 59 +/- 2 nm ) , and this was prevented by orbofiban , xemilofiban , and the monoclonal DB00054 but not by integrelin . Finally confocal microscopy showed that RGDS co-localized with caspase-3 inside the cell . These data show that RGDS and its mimetics induce cardiomyocyte apoptosis by direct activation of procaspase-3 . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . New antiplatelet agents : platelet P08514 /IIIa antagonists . The P08514 /IIIa ( alpha IIb beta 3 ) receptor plays a crucial role in platelet aggregation and platelet thrombus formation . Inhibition of P08514 /IIIa with the Fab fragment of the mouse/human chimeric monoclonal antibody DB00054 , snake venom peptides containing the arginine-glycine-aspartic acid ( RGD ) sequence , or peptides or peptidomimetics based on the RGD sequence results in abolition of platelet aggregation and platelet thrombus formation . This results in profound inhibition of thrombotic occlusions in animal models . The Phase III EPIC study demonstrated that c7E3 Fab , given as bolus followed by a 12 h infusion , reduced the risk of acute ischemic complications after coronary angioplasty by approximately 35 % in patients at high risk of suffering such complications . Treated patients had an approximately 2-fold increased risk of major bleeding , but no increase in cerebral hemorrhage or lethal bleeding . Treatment with c7E3 Fab may have had a beneficial effect on clinical restenosis at 6 months , but this needs to be confirmed . A possible anticoagulant effect of c7E3 Fab was also identified in EPIC , and in vitro studies support this possibility . With the approval of c7E3 Fab ( abciximab ; ReoPro ) for patients undergoing high-risk angioplasty in the US and several European and Scandinavian countries , P08514 /IIIa inhibition joins the armamentarium of antithrombotic agents . Time course of the effects of a single bolus injection of F(ab')2 fragments of the antiplatelet P08514 /IIIa antibody 7E3 on arterial eversion graft occlusion , platelet aggregation , and bleeding time in dogs . The time course of the effects of a single intravenous bolus injection of 10 mg/kg aspirin or 0.8 mg/kg F(ab')2 fragments of the monoclonal antiplatelet glycoprotein IIb/IIIa receptor antibody DB00054 [ 7E3-F(ab')2 ] on arterial occlusion , platelet aggregation , and bleeding time was studied in 30 dogs with an everted ( inside out ) carotid arterial segment inserted into the femoral artery . In the absence of an antiplatelet agent , the eversion grafts occluded spontaneously with platelet-rich thrombus within 30 minutes . With aspirin , arterial occlusion persisting for 2 hours occurred in 5 of 10 dogs and cyclic occlusion and reflow in 4 animals ; arterial occlusion was observed in all dogs at 24 hours . With 7E3-F(ab')2 , arterial patency persisted throughout a 2-hour observation period in all of 10 dogs and for 24 hours in 4 of the 10 dogs . Contralateral eversion grafting 24 hours after aspirin or 7E3-F(ab')2 injection was associated with graft patency for 2 hours in 1 of 5 aspirin dogs and in 3 of 5 7E3-F(ab')2 dogs ; patency persisted for 24 hours . In dogs grafted 48 hours after aspirin or 7E3-F(ab')2 injection , patency at 24 hours was seen in 0 of 5 dogs given aspirin and 3 of 5 dogs given 7E3-F(ab')2 . The overall frequencies of arterial graft patency at 2 , 24 , 48 , and 72 hours after study drug injection were significantly higher in the 7E3-F(ab')2 groups than in the aspirin groups ( P < .0005 , n = 10 in each group ; P < .05 , n = 15 ; P < .005 , n = 15 ; and P = .05 , n = 5 , respectively ) . ( ABSTRACT TRUNCATED AT 250 WORDS ) Flow cytometric analysis of platelet activation throughout normal gestation . OBJECTIVE : To measure platelet activation in normal pregnancy , before and after stimulation with agonists , with a whole blood flow cytometric technique . METHODS : In a cross-sectional study , 5 mL of whole blood was collected from healthy volunteers ( nine in the first trimester , ten in the second trimester , 35 in the third trimester , and 32 nonpregnant controls ) . Platelets were treated with an agonist ( thrombin or U-46619 , a thromboxane A2 analogue ) or buffer and were exposed to saturating concentrations of monoclonal antibodies directed against platelet membrane glycoproteins ( GPs ) : DB00054 ( fibrinogen receptor P08514 /IIIa ) , P28222 ( alpha granule marker P16109 ) , and 6D1 ( P04275 receptor GPIb ) . Mean fluorescence intensity was determined for 5000 platelets per sample by using a flow cytometer . RESULTS : In the absence of agonist , no significant difference between groups was found in antibody binding . At no stage of pregnancy were circulating activated platelets detected . Platelets from third-trimester subjects bound significantly less 7E3 than platelets of controls or of first- or second-trimester subjects after stimulation with high-dose thrombin ( P < .05 for all comparisons ) . Down-regulation of 6D1 on platelets after stimulation with high-dose U-46619 was significantly greater in third-trimester gravidas than in controls or first-trimester subjects ( P < .05 ) . CONCLUSION : Pregnancy does not increase the percentage of activated platelets in the circulation . Platelet reactivity is altered in the third trimester , as evidenced by decreased antibody binding to fibrinogen receptor epitope and enhanced down-regulation of a P04275 receptor epitope . Effects of G- P04141 on systemic inflammation , coagulation and platelet activation in patients with acute myocardial infarction . INTRODUCTION : In the prospective , randomised , double-blind , placebo-controlled Regenerate Vital Myocardium by Vigorous Activation of Bone Marrow Stem Cells ( REVIVAL ) -2 trial patients with acute myocardial infarction ( AMI ) and successful mechanical reperfusion received granulocyte-colony stimulating factor ( DB00099 , 10 μg/kg KG s.c. ) or placebo for 5 days . Aim of this substudy was to assess the impact of G- P04141 on systemic inflammatory and procoagulant responses and platelet activation . METHODS AND RESULTS : Before and five days after DB00099 ( n=56 ) or placebo ( n=58 ) circulating cytokine concentrations of interleukin ( IL ) -1ß , P05231 , P10145 , P22301 , IL-12 and Tumor-Necrosis Factor-α ( P01375 -α were measured . P00734 fragment F1+2 and Tissue Factor activity served as a measure for activated coagulation . Platelet activation was characterized by cell surface expression of the activated fibrinogen receptor ( O95456 ) , P16109 and P29965 by flow cytometry . Administration of G- P04141 was associated with elevated P01375 -α and CRP concentrations compared to the placebo group after 5 days . Other cytokines ( IL-1ß , P05231 , P10145 , P22301 , IL-12 ) were comparable after treatment with G- P21583 or placebo . Similarly , circulating prothrombin fragments F1+2 , TF activity and platelet activation did not differ in both groups . CONCLUSION : Treatment with G- P04141 in patients with AMI was associated with enhanced proinflammatory P01375 -α and CRP levels but no activation of coagulation . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Concentration-dependent effect of abciximab on platelets and neutrophils in a model of cardiopulmonary bypass . DB00054 is a P08514 /IIIa antagonist used in percutaneous coronary interventions to avoid platelet activation , thrombosis and inflammation . We investigated whether abciximab influenced platelet activation and platelet interaction with neutrophils and polyvinyl chloride ( PVC ) in a cardiopulmonary bypass ( P15086 ) model . Isolated platelets , preincubated with and without 0.1-20 microg/mL of abciximab , were resuspended with neutrophils in plasma and recirculated by a roller pump . Platelet , but not neutrophil adhesion to PVC was inhibited by abciximab . Only high doses of abciximab reduced platelet aggregation size , but simultaneously increased platelet-neutrophil aggregation . DB00054 had no effect on platelet CD62P expression or degranulation , but platelet activation on platelet-neutrophil aggregates increased with high doses . Only low doses inhibited neutrophil degranulation . The concentration-dependent effect of abciximab on platelet-neutrophil interaction reduces its usefulness and stresses the dependency on experimental design in the evaluation of abciximab . Our study does not support the use of abciximab alone in P15086 . However , incorporation of surface-coating the biomaterial with abciximab may be an interesting option . DB00054 : a reappraisal of its use in coronary care . Platelet reactivity plays a pivotal role in the pathogenesis of ischemic adverse events during and after acute coronary syndromes ( ACS ) , and percutaneous coronary intervention ( P05154 ) . Glycoprotein ( GP ) IIb/IIIa inhibitors are the strongest antiplatelet agents currently available on the market and three different compounds , namely abciximab , tirofiban , and eptifibatide , have been approved for clinical use . DB00054 has been investigated in the clinical field far more extensively than the other P08514 /IIIa inhibitors . DB00054 is an anti-integrin Fab fragment of a human - mouse chimeric monoclonal antibody with high affinity and a slow dissociation rate from the GP IIb/IIIa platelet receptor . DB00054 , given shortly before the coronary intervention , is superior to placebo in reducing the acute risk of ischemic complications ( EPIC , EPISTENT , EPILOG trials ) ; moreover , in the ISAR-REACT 2 study abciximab has been shown to reduce the risk of adverse events in patients with non ST-segment elevation ACS who are undergoing P05154 even after optimal pre-treatment with 600 mg of clopidogrel . Finally , abciximab has been also used in abciximab-coated stent , with only bolus administration regimen and for direct intracoronary use with promising results that may extend and/or modify its current use in clinical practice in future . Platelet-derived microparticle formation involves glycoprotein IIb-IIIa . Inhibition by RGDS and a Glanzmann 's thrombasthenia defect . While the physiologic role of platelet microparticles may include a stable , physical dispersion of concentrated surface procoagulant activity the mechanism(s) of platelet vesiculation remains unknown . We demonstrate using flow cytometric methods a central role for the beta 3 integrin glycoprotein ( GP ) IIb-IIIa complex and its ligand tetrapeptide DB00125 - DB00145 - DB00128 - DB00133 ( RGDS ) binding site in platelet vesiculation . Time- and calcium-dependent vesiculation of platelets in response to ADP , collagen , thrombin , phorbol myristate acetate , and the thrombin peptide SFLLRN were dramatically inhibited , in a concentration-dependent manner , by monoclonal antibodies to P08514 -IIIa ( A2A9 , DB00054 , PAC1 ) and RGDS . Complete inhibition with A2A9 and RGDS occurred at 7.5 micrograms/ml and 75 microM , respectively , while control antibodies and a mock peptide had no effect . Platelet vesiculation requires intact P08514 -IIIa and is fully supported by the intracellular pool of P08514 -IIIa alone since de-complexing of this heterodimer by calcium chelation completely abolished microparticle formation in response to collagen ( no alpha-granule release ) but not to thrombin or SFLLRN . A central role for P08514 -IIIa is supported by the near total inability of Glanzmann 's thrombasthenic ( type I ) platelets to vesiculate in response to thrombin , ADP , collagen , and phorbol 12-myristate 13-acetate . This extends the biologic roles of P08514 -IIIa to include platelet vesiculation and suggests that one or all of its binding ligands play a role . P01375 and vascular endothelial growth factor induce endothelial integrin repertories , regulating endovascular differentiation and apoptosis in a human extravillous trophoblast cell line . Angiogenesis is crucial in human development . Extravillous trophoblast ( EVT ) cells mimic endothelial cells in angiogenesis during endovascular differentiation , inducing a remodeling of spiral arteries that increases blood flow toward the intravillous space . We have previously shown that tumor necrosis factor ( P01375 ) alpha regulates expression of P23229 and P56199 , which are involved in cell survival , in the human EVT cell line TCL1 . To further investigate endovascular differentiation , we examined the effects of vascular endothelial growth factor ( P15692 ) , P01375 , and extracellular matrix ( Q13201 ) on TCL1 cells . Seeded on Matrigel , TCL1 cells show tube-like formation that specifically recalls morphological changes in endothelial cells . Anti- P06756 / P05106 antibodies significantly reduced the size of the capillary network ( P < 0.05 ) on Matrigel and also suppressed P01375 -induced apoptosis ( P < 0.05 ) in TCL1 cells . P15692 induced expression of P06756 / P05106 subunits and protein aggregation , as in the case of P01375 , which in turn , induces synthesis of P15692 in TCL1 cells . Soluble P17948 suppressed these activities in TCL1 cells , indicating that signals involving P15692 axis are essential for endovascular differentiation . These results suggest that P01375 , P15692 , and Q13201 collaboratively regulate EVT behavior , including cell survival and endovascular differentiation , through integrin signaling during establishment and maintenance of successful human pregnancies . Practical applications of snake venom toxins in haemostasis . Snake venom toxins have an established role in the coagulation laboratory for the assay of haemostatic parameters and a potential role for therapeutic treatment of thrombotic disorders . In the laboratory , snake venom thrombin-like enzymes ( SVTLEs ) are used for the assay of fibrinogen and detection of fibrinogen breakdown products and dysfibrinogenaemias . Importantly , because SVTLEs are not inhibited by heparin , they can be used for assaying antithrombin III and other parameters in samples which contain heparin . P00734 activators occur in many snake venoms and these have become established in the assay of prothrombin , in the study of dysprothrombinaemias and in the preparation of meizothrombin and non enzymic forms of prothrombin . Russell 's viper ( Daboia russelli ) venom contains a number of useful compounds including toxins which can be used to assay blood clotting factors V , VII , X , platelet factor 3 and lupus anticoagulants ( LA ) . More recently , activators from the taipan , Australian brown snake and saw-scaled viper have been used to assay LA . Proteins C and S can be measured by means of protac , a fast acting inhibitor from Southern copperhead snake venom and P04275 can be studied with botrocetin from Bothrops jararaca venom . The disintegrins , a large family of DB00125 - DB00145 - DB00128 ( RGD ) -containing proteins found in snake venoms , show great potential for the study of platelet glycoprotein receptors , notably , P08514 /IIIa and Ib , and in the treatment of arterial thrombotic disease . Established SVTLEs used in clinical practice include ancrod and defibrase although success with these agents has been limited . A further group of enzymes under consideration as thrombolytic agents are the fibrinogenases . DB00054 pharmacodynamics are unaffected by antecedent therapy with other P08514 /IIIa antagonists in non-human primates . BACKGROUND : Tirofiban and eptifibatide are currently approved for the medical stabilization of non-ST segment elevation acute coronary syndromes . In patients undergoing percutaneous coronary intervention ( P05154 ) during infusion of these drugs , conversion to abciximab , which has long term proven clinical efficacy and cost-effectiveness , following P05154 may be desirable . The purpose of this study was to determine if the binding or pharmacodynamics of abciximab is affected by a prior infusion of either tirofiban or eptifibatide . METHODS : In vitro binding experiments were performed to determine if prior exposure to tirofiban or eptifibatide altered the affinity and extent of binding of abciximab to P08514 /IIIa . For in vivo experiments , cynomolgus monkeys were pretreated with a bolus and 18 hour infusion of saline , tirofiban , or eptifibatide . At the end of the initial treatment , a bolus and 12 hr infusion of abciximab was started without delay . Inhibition of platelet aggregation , P08514 /IIIa receptor blockade and abciximab pharmacokinetics were measured during and after both infusions . RESULTS : Equilibrium binding of abciximab in vitro was unaffected by tirofiban or eptifibatide . The extent and duration of abciximab inhibition of ex vivo platelet aggregation , receptor blockade , and abciximab pharmacokinetics in monkeys during and after the abciximab infusion were not affected by prior infusion of the animals with tirofiban or eptifibatide . CONCLUSIONS : In vitro and in vivo studies revealed that the molecular interaction of abciximab with the platelet P08514 /IIIa receptor is not altered by immediate prior exposure of platelets to small molecule P08514 /IIIa antagonists . These preclinical studies suggest that the efficacy of abciximab should not be impaired if it is initiated following termination of therapy with small molecule P08514 /IIIa antagonists . Progress in the field of P08514 /IIIa antagonists . Platelet aggregation plays an important role in pathological situations such as myocardial infarction , unstable angina , peripheral artery disease , and stroke . Thus , pharmacological agents that specifically inhibit platelet aggregation are of great interest in the treatment and prevention of these cardiovascular diseases . Since binding of activated glycoprotein IIb/IIIa complex , a platelet surface integrin , to fibrinogen is the final step leading to platelet aggregation regardless of the initial stimulus , many researches have focused on the development of drugs that could antagonize this integrin . Three intravenous glycoprotein IIb/IIIa antagonists are currently marketed for the prevention of myocardial infarction in patients undergoing percutaneous intervention : DB00054 , DB00063 and Tirofiban . To further test the clinical efficacy of these agents , oral glycoprotein IIb/IIIa antagonists have been developed but only led to disappointing clinical results . Nevertheless , due to recognized usefulness of oral agents for the prevention and treatment of cardiovascular diseases , a great number of new orally active compounds are under clinical or preclinical evaluation . The aim of this review is to describe the chemical , pharmacological and clinical properties of existing and forthcoming glycoprotein IIb/IIIa antagonists . Rapid and sustained coronary artery recanalization with combined bolus injection of recombinant tissue-type plasminogen activator and monoclonal antiplatelet P08514 /IIIa antibody in a canine preparation . The effects of bolus injections of recombinant single-chain tissue-type plasminogen activator ( rt-PA ) and of F(ab')2 fragments of a murine monoclonal antibody ( DB00054 ) against the human platelet P08514 /IIIa receptor [ 7E3-F(ab')2 ] on coronary arterial thrombolysis and reocclusion was studied in a canine preparation of coronary artery thrombosis superimposed on high-grade stenosis . Bolus intravenous injections of rt-PA at a dose of 0.45 mg/kg , repeated at 15 min intervals until reperfusion occurred ( maximum of four injections ) caused reperfusion in five of seven dogs within 100 min ( 33 +/- 15 min , mean +/- SD ) . Reperfusion was rapidly followed ( generally within 10 min ) by reocclusion and then by periods of cyclical reflow and reocclusion . A single intravenous injection of 7E3-F(ab')2 alone at 0.8 mg/kg caused reperfusion within 100 min in two of six dogs ( 19 and 37 min ) without subsequent reocclusion . Single bolus injections of different amounts ( 0.1 to 0.8 mg/kg ) of 7E3-F(ab')2 were then combined with bolus injections of 0.45 mg/kg of rt-PA . Stable reperfusion without reocclusion was accomplished with 0.8 or 0.6 mg/kg 7E3-F(ab')2 and a single injection of 0.45 mg/kg rt-PA within 6 +/- 3 min ( n = 6 , p less than .01 ) and 8 +/- 5 min ( n = 5 , p less than .02 ) , respectively . None of these animals suffered reocclusion of the coronary artery . Lower doses ( 0.1 to 0.2 mg/kg ) of 7E3-F(ab')2 did not significantly shorten the time to reperfusion and did not prevent reocclusion. ( ABSTRACT TRUNCATED AT 250 WORDS ) Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome .

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DB00398 inhibits the angiogenesis and growth of orthotopic anaplastic thyroid carcinoma xenografts in nude mice . Anaplastic thyroid carcinoma ( ATC ) remains one of the most lethal human cancers . We hypothesized that sorafenib , a multikinase inhibitor of the BRaf , vascular endothelial growth factor receptor-2 , and platelet-derived growth factor receptor-beta kinase , would decrease tumor growth and angiogenesis in an orthotopic model of ATC . The in vitro antiproliferative and proapoptotic effects of sorafenib on ATC cell lines were examined . To study the in vivo effects of sorafenib on orthotopic ATC tumors in nude mice , sorafenib was given p.o. at 40 or 80 mg/kg daily . Intratumoral effects were studied using immunohistochemical analysis . The effect of sorafenib on survival of the mice was also studied . DB00398 inhibited the in vitro proliferation of ATC cell lines . DB00398 also significantly inhibited tumor angiogenesis via the induction of endothelial apoptosis in an orthotopic model of thyroid cancer . As result , the growth of orthotopic ATC xenografts was reduced and the survival of the test animals was improved . DB00398 exerts significant antitumor activity in an orthotopic xenograft model of ATC via a potent antiangiogenic effect . The antiangiogenic effects of sorafenib suggest that its use in clinical setting may not depend on the P15056 mutational status of thyroid tumors . Given the lack of curative options for patients with ATC , sorafenib warrants further study as a therapeutic agent against ATC . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . Synthesis , biological evaluation and 3D-QSAR studies of novel 4,5-dihydro-1H-pyrazole niacinamide derivatives as P15056 inhibitors . A series of novel 4,5-dihydropyrazole derivatives containing niacinamide moiety as potential V600E mutant P15056 kinase ( P15056 (V600E) ) inhibitors were designed and synthesized . Results of the bioassays against P15056 (V600E) and WM266.4 human melanoma cell line showed several compounds to be endowed potent activities with IC(50) and GI(50) value in low micromolar range , among which compound 27e , (5-(4-Chlorophenyl)-3-(4-methoxyphenyl)-4,5-dihydro-1H-pyrazol-1-yl)6-methylpyridin-3-yl methanone ( IC(50)=0.20 μM , GI(50)=0.89 μM ) was bearing the best bioactivity comparable with the positive control DB00398 . Docking simulation was performed to determine the probable binding model and 3D-QSAR model was built to provide more pharmacophore understanding that could use to design new agents with more potent P15056 (V600E) inhibitory activity . Molecular targeted therapies for cancer : sorafenib mono-therapy and its combination with other therapies ( review ) . DB00398 is an oral multikinase inhibitor that acts by inhibiting tumor growth and disrupting tumor microvasculature through antiproliferative , anti-angiogenic and proapoptotic effects . It exerts these effects via inhibition of multiple targets including Raf serine/threonine kinases , vascular endothelial growth factor receptor tyrosine kinases ; P17948 , P35968 , P35916 and platelet-derived growth factor receptor β ( P09619 -β ) . Current literature shows that the deregulated signaling through these receptors is commonly seen in human tumors . In addition , sorafenib is also shown to induce apoptosis through downregulation of Mcl-1 in many cancer types . Hence , sorafenib as a single agent has shown promising activity in some cancers such as renal cell carcinoma ( RCC ) , hepatocellular carcinoma ( HCC ) and thyroid cancers . Currently , the drug holds FDA approval for the treatment of advanced RCC and unresectable HCC . However , many clinical studies have indicated several limitations to the application of sorafenib as a single agent in various other cancers . Owing to these reasons and the potential of sorafenib to synergize with other anticancer therapies , its combination with other targeted agents and chemotherapy has been widely explored with promising results . In addition , it has also shown synergistic results when combined with radiation . This review summarizes the current basic and clinical studies on the effects and mechanisms of sorafenib either administered alone or in combination with other anticancer treatments . NFκB inhibitors induce cell death in glioblastomas . Identification of novel target pathways in glioblastoma ( GBM ) remains critical due to poor prognosis , inefficient therapies and recurrence associated with these tumors . In this work , we evaluated the role of nuclear-factor-kappa-B ( NFκB ) in the growth of GBM cells , and the potential of NFκB inhibitors as antiglioma agents . NFκB pathway was found overstimulated in GBM cell lines and in tumor specimens compared to normal astrocytes and healthy brain tissues , respectively . Treatment of a panel of established GBM cell lines ( U138MG , U87 , U373 and P13671 ) with pharmacological NFκB inhibitors ( BAY117082 , parthenolide , MG132 , curcumin and arsenic trioxide ) and NFκB-p65 siRNA markedly decreased the viability of GBMs as compared to inhibitors of other signaling pathways such as MAPKs ( P29323 , JNK and p38 ) , PKC , P00533 and PI3K/Akt . In addition , NFκB inhibitors presented a low toxicity to normal astrocytes , indicating selectivity to cancerous cells . In GBMs , mitochondrial dysfunction ( membrane depolarization , bcl-xL downregulation and cytochrome c release ) and arrest in the G2/M phase were observed at the early steps of NFκB inhibitors treatment . These events preceded sub- P55008 detection , apoptotic body formation and caspase-3 activation . Also , NFκB was found overstimulated in cisplatin-resistant P13671 cells , and treatment of GBMs with NFκB inhibitors overcame cisplatin resistance besides potentiating the effects of the chemotherapeutics , cisplatin and doxorubicin . These findings support NFκB as a potential target to cell death induction in GBMs , and that the NFκB inhibitors may be considered for in vivo testing on animal models and possibly on GBM therapy . Induction of apoptosis in P11274 / P00519 + cells by histone deacetylase inhibitors involves reciprocal effects on the RAF/MEK/ P29323 and JNK pathways . Signal transduction events regulating induction of apoptosis by the histone deacetylase inhibitors ( HDIs ) sodium butyrate ( SB ) and DB02546 have been examined in Bcr/Abl+ human leukemia cells ( K562 , P25391 84 ) . Exposure of K562 cells to greater or less than 3.0 mM SB or 3.0 mM DB02546 for 24-48 hr resulted in a marked induction of mitchondrial damage ( e.g. , cytochrome c release ) and apoptosis , events associated with downregulation of Bcr/Abl and P04049 , induction of p21CIP1 , inactivation of Q02750 /2 , P27361 /2 , and p70S6K , and a dramatic increase in JNK activation . HDI-mediated apoptosis was attenuated by pharmacologic JNK inhibitors and enhanced by the Q02750 /2 inhibitor U0126 as well as by the JNK activator anisomycin . Interestingly , HDI-induced JNK activation was potentiated by pharmacologic MEK inhibition . Furthermore , HDI lethality was significantly diminished in cells ectopically expressing constitutively active Q02750 , confirming a functional role for MEK/ P29323 inactivation in HDI-mediated apoptosis . Similar events were observed in Bcr/Abl+ P25391 84 cells . Lastly , the free radical scavenger L- DB06151 ( LNAC ) attenuated HDI-mediated ROS generation , JNK activation , and apoptosis . Together , these findings support a model in which induction of apoptosis in Bcr/Abl+ cells by HDIs involves coordinate inactivation of the cytoprotective Raf/MEK/ P29323 pathway in conjunction with the ROS-dependent activation of JNK . Survey and analysis of the efficacy and prescription pattern of sorafenib in patients with acute myeloid leukemia . DB00398 is a multi-kinase inhibitor with activity against several intracellular kinases which may play a role in the pathogenesis of acute myeloid leukemia ( AML ) . In vitro data and results from early clinical trials suggest that sorafenib might be an effective drug for the treatment of AML . However , clinical data are still sparse , and there are only a few reported cases of monotherapy . The aim of the present research was to collect clinical data on efficacy and safety in a systematic way by conducting a survey on clinical experience with sorafenib . Thirty institutions were asked to document all patients treated with sorafenib diagnosed with AML . Of all 29 evaluable patients , six ( 21 % ) responded to sorafenib containing treatment by achieving a complete remission ( CR , n = 2 ) or complete remission with incomplete platelet recovery ( CRi , n = 4 ) . In 23 patients receiving sorafenib as monotherapy , the CRi rate amounted to 13 % and no CRs were documented . Of the 18 P17948 -ITD positive patients with sorafenib monotherapy , two patients achieved a CRi ( 11 % ) . In five P36888 -ITD negative cases , one CRi was documented ( 20 % ) . Our results suggest the potential ability of the drug to induce remissions in refractory or relapsed AML even when given as monotherapy . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . A novel tescalcin-sodium/hydrogen exchange axis underlying sorafenib resistance in P36888 -ITD+ AML . Internal tandem duplication ( ITD ) of fms-like tyrosine kinase 3 ( P36888 ) in acute myeloid leukemia ( AML ) is associated with inferior clinical prognosis . DB00398 is effective in clearing leukemic blasts in chemorefractory P36888 -ITD(+) AML , but leukemia progression invariably occurs . Mechanisms of drug resistance are not completely understood . We hypothesized that a gene encoding tescalcin ( Q96BS2 ) , known to be upregulated at leukemia progression during continuous sorafenib treatment and activate an Na(+)/H(+) exchanger type-1 ( P19634 ) , may underlie tyrosine kinase inhibitor resistance . Q96BS2 was highly expressed in P36888 -ITD(+) AML lines MOLM-13 and MV4-11 , and its knockdown by small-interfering RNA lowered intracellular pH ( pHi ) and induced apoptosis . The results were recapitulated by treatment with an P19634 inhibitor , 5-(N,N-hexamethylene) amiloride ( HMA ) . Induction of sorafenib resistance in the MOLM-13 cell line ( M13-RE ) significantly increased its sensitivity to HMA . The later also enhanced suppression of P36888 signaling by sorafenib in otherwise resistant cell lines . HMA treatment of MOLM-13 and MV4-11 as well as primary P36888 -ITD(+) AML cells significantly reduced leukemia initiation in anti-CD122-primed NOD/SCID mouse xenotransplantation . These observations provided novel information about the pathogenetic role of a Q96BS2 - P19634 -pHi axis in mediating sorafenib resistance in AML . Renal cell carcinoma and the use of sorafenib . Immunotherapy results in a small overall survival advantage in metastatic renal cell carcinoma ( RCC ) , but there is a need to develop more effective systemic therapies . Angiogenesis has an important role in the pathophysiology of RCC and vascular endothelial growth factor ( P15692 ) is a key mediator of this process . DB00398 ( BAY 43-9006 ) is a new agent belonging to a class of drugs called kinase inhibitors and inhibits the P15692 , platelet-derived growth factor ( PDGF ) , and c- P10721 receptor tyrosine kinases , amongst others . DB00398 has shown significant activity with manageable toxicity in metastatic RCC in phase 2 studies in patients pretreated with immunotherapy , whilst prolonged progression-free survival in comparison with placebo in a phase 3 study has been reported . Further phase 3 trials in advanced disease are ongoing and a trial of adjuvant sorafenib therapy in RCC is planned . DB00398 inhibits nuclear factor kappa B , decreases inducible nitric oxide synthase and cyclooxygenase-2 expression , and restores working memory in APPswe mice . Alzheimer 's disease ( AD ) is characterized by memory loss and the upregulation of pro-neuroinflammatory factors such as P04049 -1 , cyclooxygenase-2 ( Cox-2 ) , and the nuclear factor kappa B ( NF-kappaB ) , as well as a downregulation of protein kinase A ( PKA ) activity and the activation by phosphorylation of its downstream factor CREB . We investigated the effect of the anti-cancer P04049 -1 inhibitor , sorafenib tosylate ( Nexavar ) , on the expression of these factors and on the cognitive performance of aged APPswe mice . We found that chronic treatment with sorafenib stimulated PKA and CREB phosphorylation and inhibited P04049 -1 and NF-kappaB in the brains of APPswe mice . NF-kappaB controls the expression of several genes related to AD pathology , including P35228 and Cox-(2)Concurrent with NF-kappaB inhibition , sorafenib treatment decreased the cerebral expression of Cox-2 and P35228 in APPswe mice . It has recently been observed that Cox-2 inhibition prevents cognitive impairment in a mouse model of AD and amyloid beta peptide ( Abeta ) -induced inhibition of long-term potentiation ( LTP ) . Consistent with the idea that Cox-2 inhibition can improve cognitive abilities , we found that sorafenib restored working memory abilities in aged APPswe mice without reducing Abeta levels in the brain . These findings suggest that sorafenib reduced AD pathology by reducing neuroinflammation . Immunohistochemical study of P15692 , angiopoietin 2 and their receptors in the neovascularization following microinjection of P13671 glioma cells into rat brain . BACKGROUND : Recent papers suggest that two angiogenic factors ( angiopoietin 2 and vascular endothelial growth factor ) cooperate in tumoral angiogenesis to support the growing tumor . The purpose of the present work was to demonstrate the existence of such cooperation in a longitudinal study of a brain tumor model during tumor growth by means of immunohistochemistry . MATERIALS AND METHODS : The study was performed on 31 rats bearing P13671 glioma . At different stages of tumor growth , the histological aspects were described and sections were immunostained for P15692 , Ang-2 and their receptors P17948 , P35968 and Tie-2 . Immunostaining was semi-quantitatively analyzed and the localization of immunostained cells was reported . RESULTS : Ang-2 and Tie-2 were detected in the endothelial cells of vessels surrounded by tumor cells , occuring early in our study , with immunostaining taking place from day 4 to day 24 . Immunostaining with P15692 ( on tumoral cells ) and P35968 ( on endothelial cells ) was present after 8 days of tumor growth . A clear increase of vessel density can be observed at the periphery of the tumors after 16 days of tumor growth . At that time , areas of necrosis were present in the tumor with concomitant P15692 and Ang-2 expression . CONCLUSION : The present study demonstrated cooperation between the early effect of Ang-2 and the secondary effect of P15692 to elaborate new vessels in a longitudinal study of experimental brain tumors . This study is favorable to the new model of tumor angiogenesis , with successive vessel cooption , regression and growth mediated by angiopoietins and P15692 . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . [ DB00398 monotherapy for P36888 -ITD positive acute myeloid leukemia : a case report ] . Indispensable functions of P00519 and PDGF receptor kinases in epithelial adherence of attaching/effacing pathogens under physiological conditions . Enteropathogenic Escherichia coli ( EPEC ) and Citrobacter rodentium are attaching-and-effacing ( A/E ) pathogens that cause intestinal inflammation and diarrhea . The bacteria adhere to the intestinal epithelium , destroy microvilli , and induce actin-filled membranous pedestals but do not invade the mucosa . Adherence leads to activation of several host cell kinases , including P06241 , n- P12931 , P07947 , P00519 , and ARG , phosphorylation of the bacterial translocated intimin receptor , and actin polymerization and pedestal formation in cultured cells . However , marked functional redundancy appears to exist between kinases , and their physiological importance in A/E pathogen infections has remained unclear . To address this question , we employed a novel dynamic in vitro infection model that mimics transient and short-term interactions in the intestinal tract . Screening of a kinase inhibitor library and RNA interference experiments in vitro revealed that P00519 and platelet-derived growth factor ( PDGF ) receptor ( P09619 ) kinases , as well as p38 Q96HU1 kinase , have unique , indispensable roles in early attachment of EPEC to epithelial cells under dynamic infection conditions . Studies with mutant EPEC showed that the attachment functions of P00519 and P09619 were independent of the intimin receptor but required bacterial bundle-forming pili . Furthermore , inhibition of P00519 and P09619 with imatinib protected against infection of mice with modest loads of C. rodentium , whereas the kinases were dispensable for high inocula or late after infection . These results indicate that P00519 and P09619 have indispensable roles in early A/E pathogen attachment to intestinal epithelial cells and for in vivo infection with limiting inocula but are not required for late intimate bacterial attachment or high inoculum infections . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . P15056 inhibition alleviates immune suppression in murine autochthonous melanoma . A growing body of evidence suggests that P15056 inhibitors , in addition to their acute tumor growth-inhibitory effects , can also promote immune responses to melanoma . The present study aimed to define the immunologic basis of P15056 -inhibitor therapy using the Braf/Pten model of inducible , autochthonous melanoma on a pure C57BL/6 background . In the tumor microenvironment , P15056 inhibitor PLX4720 functioned by on-target mechanisms to selectively decrease both the proportions and absolute numbers of P01730 (+)Foxp3(+) regulatory T cells ( Treg ) and CD11b(+)Gr1(+) myeloid-derived suppressor cells ( MDSC ) , while preserving numbers of CD8(+) effector T cells . In PLX4720-treated mice , the intratumoral Treg populations decreased significantly , demonstrating enhanced apopotosis . CD11b(+) myeloid cells from PLX4720-treated tumors also exhibited decreased immunosuppressive function on a per-cell basis . In accordance with a reversion of tumor immune suppression , tumors that had been treated with PLX4720 grew with reduced kinetics after treatment was discontinued , and this growth delay was dependent on CD8 T cells . These findings demonstrate that P15056 inhibition selectively reverses two major mechanisms of immunosuppression in melanoma and liberates host-adaptive antitumor immunity . DB00398 in combination with intensive chemotherapy in elderly patients with acute myeloid leukemia : results from a randomized , placebo-controlled trial . PURPOSE : The prognosis of elderly patients with acute myeloid leukemia ( AML ) is still dismal even with intensive chemotherapy . In this trial , we compared the antileukemic activity of standard induction and consolidation therapy with or without the addition of the kinase inhibitor sorafenib in elderly patients with AML . PATIENTS AND METHODS : All patients received standard cytarabine and daunorubicin induction ( 7+3 regimen ) and up to two cycles of intermediate-dose cytarabine consolidation . Two hundred one patients were equally randomly assigned to receive either sorafenib or placebo between the chemotherapy cycles and subsequently for up to 1 year after the beginning of therapy . The primary objective was to test for an improvement in event-free survival ( O43281 ) . Overall survival ( OS ) , complete remission ( CR ) rate , tolerability , and several predefined subgroup analyses were among the secondary objectives . RESULTS : Age , sex , CR and early death ( ED ) probability , and prognostic factors were balanced between both study arms . Treatment in the sorafenib arm did not result in significant improvement in O43281 or OS . This was also true for subgroup analyses , including the subgroup positive for P36888 internal tandem duplications . Results of induction therapy were worse in the sorafenib arm , with higher treatment-related mortality and lower CR rates . More adverse effects occurred during induction therapy in the sorafenib arm , and patients in this arm received less consolidation chemotherapy as a result of higher induction toxicity . CONCLUSION : In conclusion , combination of standard induction and consolidation therapy with sorafenib in the schedule investigated in our trial is not beneficial for elderly patients with AML . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Expression of the adaptor protein Lnk in leukemia cells . OBJECTIVE : Tyrosine kinases are involved in cytokine signaling and are frequently aberrantly activated in hematological malignancies . Lnk , a negative regulator of cytokine signaling , plays critical nonredundant roles in hematopoiesis . By binding to phosphorylated tyrosine kinases , Lnk inhibits major cytokine receptor signaling , including c- P10721 ; erythropoietin receptor- O60674 ( O60674 ) ; and P40238 - O60674 . In the present study , we investigated Lnk expression and possible function in transformed hematopoietic cells . MATERIALS AND METHODS : Coimmunoprecipitations were performed to identify binding between Lnk and mutant tyrosine kinases . Proliferation assays were done to examine the affect of Lnk overexpression on cancer cell growth . Real-time polymerase chain reaction analysis was used to determine Lnk expression in patient samples . RESULTS : We show that , in parallel to binding wild-type O60674 and c- P10721 , Lnk associates with and is phosphorylated by mutant alleles of O60674 and c- P10721 . In contrast , Lnk does not bind to and is not phosphorylated by P11274 - P00519 fusion protein . Ectopic expression of Lnk strongly attenuates growth of some leukemia cell lines , while others as well as most solid tumor cancer cell lines are either moderately inhibited or completely insensitive to Lnk . Furthermore , Lnk-mediated growth inhibition is associated with differential downregulation of phosphatidylinositol 3 kinase/Akt/mammalian target of rapamycin and mitogen-activated protein kinase/extracellular signal-regulated kinase signaling in leukemia cell lines . Surprisingly , analysis of Lnk in a large panel of myelodysplastic syndrome and acute myeloid leukemia patient samples revealed high levels of Lnk in nearly half of the samples . CONCLUSION : Although how leukemic cells overcome the antiproliferative effects of Lnk is not yet clear , our data highlight the multifaceted role negative feedback mechanisms play in malignant transformation . A significant response to sorafenib in a woman with advanced lung adenocarcinoma and a P15056 non-V600 mutation . Lung adenocarcinoma includes recurrent activating oncogenic mutations ( P00533 , Q9HC35 - Q9UM73 , P08922 ) that have been associated with response to P00533 and Q9UM73 inhibitors . Platinum-based chemotherapy is the standard therapy for non-oncodrivers population . DB00398 is a small molecule that blocks the activation of C-RAF , B-RAF , c- P10721 , P36888 , P07949 , P35968 , P35916 and P09619 approved for advanced renal cell and hepatocellular carcinoma ( b , c ) . Many studies have evaluated sorafenib in advanced non-small-cell lung cancer ( NSCLC ) , with different results . We present a case report of a patient with NSCLC and the P15056 G469R mutation who showed a dramatic response to sorafenib . Role of Raf kinase in cancer : therapeutic potential of targeting the Raf/MEK/ P29323 signal transduction pathway . Improvements in our understanding of the molecular basis of cancer have led to the clinical development of protein kinase inhibitors , which target pivotal molecules involved in intracellular signaling pathways implicated in tumorigenesis and progression . These novel targeted agents have demonstrated activity against a wide range of solid tumors , are generally better tolerated than standard chemotherapeutics , and may revolutionize the management of advanced refractory cancer . The ubiquitous Raf serine/threonine kinases are pivotal molecules within the Raf/mitogen extracellular kinase ( MEK ) /extracellular signal-related kinase ( P29323 ) signaling pathway , which regulates cellular proliferation and survival . Raf kinase isoforms ( wild-type P04049 or the b-raf V600E oncogene ) are overactivated in a variety of solid tumor types , including renal cell carcinoma ( RCC ) , hepatocellular carcinoma ( HCC ) , non-small cell lung cancer ( NSCLC ) , melanoma , and papillary thyroid carcinoma . In this review , the role of Raf in normal cells and in cancer is discussed , and an overview is given of Raf inhibitors currently in development , focusing on sorafenib tosylate ( BAY 43-9006 or sorafenib ) . DB00398 is the first oral multi-kinase inhibitor to be developed that targets Raf kinases ( P04049 , wild-type B-Raf , and b-raf V600E ) , in addition to receptor tyrosine kinases associated with angiogenesis ( vascular endothelial growth factor receptor [ VEGFR ] -2/-3 , platelet-derived growth factor receptor [ P09619 ] -beta ) or tumor progression ( Flt-3 , c-kit ) . Preclinical and clinical sorafenib data that led to its recent approval for the treatment of advanced RCC are summarized , along with current thinking on sorafenib 's mechanism of effect on the tumor and tumor vasculature in melanoma and RCC . Perivascular nitric oxide activates notch signaling and promotes stem-like character in PDGF-induced glioma cells . P29474 expression is elevated in human glioblastomas and correlated with increased tumor growth and aggressive character . We investigated the potential role of nitric oxide ( NO ) activity in the perivascular niche ( PVN ) using a genetic engineered mouse model of PDGF-induced gliomas . P29474 expression is highly elevated in tumor vascular endothelium adjacent to perivascular glioma cells expressing P48681 , Notch , and the NO receptor , sGC . In addition , the NO/cGMP/PKG pathway drives Notch signaling in PDGF-induced gliomas in vitro , and induces the side population phenotype in primary glioma cell cultures . NO also increases neurosphere forming capacity of PDGF-driven glioma primary cultures , and enhances their tumorigenic capacity in vivo . Loss of NO activity in these tumors suppresses Notch signaling in vivo and prolongs survival of mice . This mechanism is conserved in human P09619 amplified gliomas . The NO/cGMP/PKG pathway 's promotion of stem cell-like character in the tumor PVN may identify therapeutic targets for this subset of gliomas . Program death-1 engagement upon TCR activation has distinct effects on costimulation and cytokine-driven proliferation : attenuation of Q9Y6W8 , P05112 , and Q9HBE4 , but not P10747 , P13232 , and P40933 responses . The program death 1 ( P18621 ) receptor and its ligands , P18621 ligand (PD-L)1 and Q9BQ51 , define a novel regulatory pathway with potential inhibitory effects on T , B , and monocyte responses . In the present study , we show that human P01730 (+) T cells express P18621 , Q9NZQ7 , and Q9BQ51 upon activation , and Abs to the receptor can be agonists or antagonists of the pathway . Under optimal conditions of stimulation , Q9Y6W8 but not P10747 costimulation can be prevented by P18621 engagement . P60568 levels induced by costimulation are critical in determining the outcome of the P18621 engagement . Thus , low to marginal P60568 levels produced upon Q9Y6W8 costimulation account for the greater sensitivity of this pathway to P18621 -mediated inhibition . Interestingly , exogenous P60568 , P13232 , and P40933 but not P05112 and Q9HBE4 can rescue P18621 inhibition , suggesting that among these cytokines only those that activate P42229 can rescue P18621 inhibition . As P42229 has been implicated in the maintenance of IL-2Ralpha expression , these results suggest that P13232 and P40933 restore proliferation under conditions of P18621 engagement by enhancing high-affinity IL-2R expression and hence , P60568 responsiveness . GPCRs promote the release of zinc ions mediated by P29475 /NO and the redox transducer Q9UGC6 protein . AIMS : Morphine signaling via the μ-opioid receptor ( MOR ) is coupled to redox-dependent zinc release from endogenous stores . Thus , MOR activation stimulates the complex formed by Q9UGC6 ( a regulator of G protein signaling ) and neural nitric oxide synthase ( P29475 ) to produce NO , and to recruit PKCγ and P04049 in a zinc-dependent manner . Accordingly , we investigated whether redox regulation of zinc metabolism was unique to the MOR , or if it is a signaling mechanism shared by G-protein coupled receptors ( GPCRs ) . RESULTS : A physical interaction with the Q9UGC6 - P29475 complex was detected for the following GPCRs : neuropeptides , MOR and δ-opioid ( Q8IXH6 ) ; biogenic amines , 5HT1A , 5HT2A , α2A , D1 and D2 ; acetylcholine , muscarinic M2 and M4 ; excitatory amino acid glutamate , mGlu2 and mGlu5 ; and derivatives of arachidonic acid ( anandamide ) , P21554 . Agonist activation of these receptors induced the release of zinc ions from the Q9UGC6 zinc finger via a P29475 /NO-dependent mechanism , recruiting PKCγ and P04049 to the C terminus or the third internal loop of the GPCR . INNOVATION : A series of GPCRs share an unexpected mechanistic feature , the P29475 /NO-dependent regulation of zinc ion signaling via a redox mechanism . The Q9UGC6 protein emerges as a potential redox zinc switch that converts NO signals into zinc signals , thereby able to modulate the function of redox sensor proteins like PKCγ or P04049 . CONCLUSION : Redox mechanisms are crucial for the successful propagation of GPCR signals in neurons . Thus , dysfunctions of GPCR-regulated NO/zinc signaling may contribute to neurodegenerative and mood disorders such as Alzheimer 's disease and depression . Optimized protocols for generation of cord blood-derived cytokine-induced killer/natural killer cells . The efficacy of various combinations of stem cell factor ( P21583 ) , P36888 ligand , interleukin ( IL ) -2 , P13232 and P40933 to induce and expand cord blood-derived cytokine-induced killer ( CIK ) cells was investigated . There were three treatment groups : group A : P21583 combined with P60568 , P13232 and P40933 ; group B : P21583 , P36888 ligand combined with P60568 , P13232 and P40933 , and group C : P60568 , P13232 and P40933 , the control group . Proliferation rates of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) natural killer ( NK ) cells were highest in group B ; expansion of CIK cells increased 796.1 ± 278.5-fold , and that of NK cells increased 36.6 ± 3.5-fold . All expanded cord blood-derived CIK/NK cells showed cytotoxic activity against the K562 cell line . Interestingly , the cytotoxicity of group A was highest and significantly higher than that of other groups . These protocols might provide an alternative choice for CIK/NK cell expansion . Personalized medicine and pharmacogenetic biomarkers : progress in molecular oncology testing . In the field of oncology , clinical molecular diagnostics and biomarker discoveries are constantly advancing as the intricate molecular mechanisms that transform a normal cell into an aberrant state in concert with the dysregulation of alternative complementary pathways are increasingly understood . Progress in biomarker technology , coupled with the companion clinical diagnostic laboratory tests , continue to advance this field , where individualized and customized treatment appropriate for each individual patient define the standard of care . Here , we discuss the current commonly used predictive pharmacogenetic biomarkers in clinical oncology molecular testing : P15056 V600E for vemurafenib in melanoma ; Q9HC35 - Q9UM73 for crizotinib and P00533 for erlotinib and gefitinib in non-small-cell lung cancer ; P01116 against the use of cetuximab and panitumumab in colorectal cancer ; P04626 ( P04626 /neu ) for trastuzumab in breast cancer ; P11274 - P00519 for tyrosine kinase inhibitors in chronic myeloid leukemia ; and P29590 /RARα for all-trans-retinoic acid and arsenic trioxide treatment for acute promyelocytic leukemia . The novel Chk1 inhibitor MK-8776 sensitizes human leukemia cells to HDAC inhibitors by targeting the intra-S checkpoint and DNA replication and repair . Interactions between the novel Chk1 inhibitor MK-8776 and the histone deacetylase ( HDAC ) inhibitor ( HDACI ) vorinostat were examined in human leukemia cells harboring wild-type ( wt ) or deficient p53 . MK-8776 synergistically potentiated vorinostat-mediated apoptosis in various p53-wt or -deficient leukemia cell lines , whereas p53 knockdown by short hairpin RNA ( shRNA ) sensitized p53-wt cells to lethality of this regimen . Leukemia cell lines carrying P36888 -ITD were also sensitive to the MK-8776/vorinostat regimen . Synergistic interactions were associated with inhibition of Chk1 activity , interference with the intra-S-phase checkpoint , disruption of DNA replication , and downregulation of proteins involved in DNA replication ( e.g. , Cdt1 ) and repair ( e.g. , Q99708 and P38398 ) , resulting in sharp increases in DNA damage , reflected by enhanced γ-H2A.X formation , and apoptosis . Moreover , leukemia cells expressing kinase-dead Chk1 ( D130A ) or Chk1 shRNA were significantly more sensitive to HDACIs compared with their wt counterparts and displayed downregulation of Q99708 and P38398 phosphorylation following HDACI exposure . Finally , the MK-8776/vorinostat regimen was active in primary acute myelogenous leukemia ( AML ) blasts , particularly against the P28906 (+)/ P28907 (-)/CD123(+) population enriched for leukemia-initiating cells . In contrast , identical regimens were relatively sparing toward normal cord blood P28906 (+) cells . Together , these findings indicate that the novel Chk1 inhibitor MK-8776 markedly potentiates HDACI lethality in leukemia cells displaying various genetic backgrounds through mechanisms involving disruption of the intra-S checkpoint , DNA replication , and DNA repair . They also argue that leukemic cells , including those bearing oncogenic mutations associated with poor prognosis , for example , p53 deletion/mutation or P36888 -ITD , may also be susceptible to this strategy . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . DB01076 attenuates bleomycin-induced pulmonary fibrosis via suppressing P35228 expression and the P29279 ( P29279 ) / P29323 signaling pathway . Pulmonary fibrosis is a progressive and fatal lung disorder with high mortality rate . To date , despite the fact that extensive research trials are ongoing , pulmonary fibrosis continues to have a poor response to available medical therapy . Statins , P04035 inhibitors , known for its broad pharmacological activities , remains a remedy against multiple diseases . The present study investigated the antifibrotic potential of atorvastatin against bleomycin-induced lung fibrosis and to further explore the possible underlying mechanisms . Our results showed that atorvastatin administration significantly ameliorated the bleomycin mediated histological alterations and blocked collagen deposition with parallel reduction in the hydroxyproline level . DB01076 reduced malondialdehyde ( MDA ) level and lung indices . DB01076 also markedly decreased the expression of inducible nitric oxide synthase ( P35228 ) in lung tissues and , thus , prevented nitric oxide ( NO ) release in response to bleomycin challenge . Furthermore , atorvastatin exhibited target down-regulation of connective tissue growth factor ( P29279 ( P29279 ) ) and phosphorylation extracellular regulated protein kinases ( p- P29323 ) expression . Taken together , atorvastatin significantly ameliorated bleomycin-induced pulmonary fibrosis in rats , via the inhibition of P35228 expression and the P29279 ( P29279 ) / P29323 signaling pathway . The present study provides evidence that atorvastatin may be a potential therapeutic reagent for the treatment of lung fibrosis . DB00398 treatment in 13 patients with acute myeloid leukemia and activating P36888 mutations in combination with chemotherapy or as monotherapy . DB00398 induces sustained molecular remission in P36888 -ITD positive AML with relapse after second allogeneic stem cell transplantation without exacerbation of acute GVHD : a case report . Potential mechanism for endothelial progenitor cell therapy in acute myocardial infarction : Activation of P15692 - PI3K/Akte-NOS pathway . Mounting evidence suggests that transplanting endothelial progenitor cells ( EPCs ) into the myocardium improves cardiac function after myocardial infarction ( MI ) . However , the mechanism remains controversial . The aim of this study was to investigate the role played by the P15692 - PI3K/Akt- P29474 pathway in EPC-based cell therapy . Cultured EPCs , which were identified by morphology , function , and cell surface markers , were transplanted into the border zone after left anterior descending coronary artery ligation in mice . Expression levels of P15692 , p-Akt , and P29474 in the border zone were elevated three days after EPC transplantation . EPC therapy enhanced expression of P35968 , increased microvessel density , and reduced interstitial fibrosis in the border zone after MI . The left ventricular fractional shortening was increased and the left ventricular diameter was smaller after EPC treatment . Wortmannin inhibited the expression of p-Akt and was associated with decreased cardiac function . Our study suggests that EPC transplantation improves cardiac function after MI , mediated at least partially by activation of the P15692 -PI3K/Akt- P29474 pathway . Novel quinolinylaminoisoquinoline bioisosteres of sorafenib as selective P04049 kinase inhibitors : design , synthesis , and antiproliferative activity against melanoma cell line . Design and synthesis of a new series of quinolinylaminoisoquinoline derivatives as conformationally restricted bioisosteres of DB00398 are described . Their in vitro antiproliferative activity against A375P melanoma cell line was tested . Compounds 1b , 1d , 1g , and 1j showed the highest potency against A375P cell line with IC50 values in sub-micromolar scale . In addition , compound 1d exerted high selectivity towards P04049 serine/threonine kinase with 96.47 % inhibition at 10 µM , and IC50 of 0.96 µM . This compound can possess antiproliferative activity against melanoma cells through inhibition of P04049 kinase . A novel tissue model for angiogenesis : evaluation of inhibitors or promoters in tissue level . A novel tissue model for angiogenesis ( TMA ) is established for effective evaluation of angiogenesis inhibitors or promoters in vitro . Lung tissues were cultured in fibrinogen " sandwich " structure which resembled the formation of neovessels in vivo . The cells and capillary-like structures grew from the lung tissues were identified as endothelial cells and neovessels . Both immunohistochemisty and western blot results indicated that autocrine P15692 bound to the P35968 and induced P35968 autophosphorylation that could induce the proliferation of endothelial cells and their migration as well as the formation of microvessels on the lung tissue edge . With addition of the TMA , the murine P15692 and cultured medium produced by A549 tumor cells apparently promoted the increase of neovessels . DB00398 as a tumor angiogenesis inhibitor and Tongxinluo as an angiogenesis promoter were both used to evaluate the TMA performance and they exhibited a good effect on neovessels in the TMA . The model established imitated angiogenesis in vivo and could well serve as an effective method in evaluating the angiogenesis inhibitors or promoters , and could also be practical for screening small molecules that affect blood vessel formation . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Synthesis and biological evaluation of novel thieno[2,3-d]pyrimidine-based P36888 inhibitors as anti-leukemic agents . The most common mutations in acute myeloid leukemia ( AML ) are those that cause the activation of P07333 -like tyrosine kinase 3 ( P36888 ) . Therefore , P36888 is regarded as a potential target for the treatment of AML . A novel series of thieno[2,3-d]pyrimidine-based analogs was designed and synthesized as P36888 inhibitors . All synthesized compounds were assayed for the tyrosine kinase activity of P36888 and growth inhibitory activity in four human leukemia cell lines ( THP1 , MV4-11 , K562 , and HL-60 ) . Among these compounds , compound 17a , which possesses relatively short and simple substituents at the P13671 position of thieno[2,3-d]pyrimidine , emerged as the most promising anti-leukemic agent . Compound 17a exhibited potent inhibition of P36888 -positive leukemic cell growth and of the P36888 D835Y kinase ; such inhibition is required for the successful treatment of AML . The data supports the further investigation of this class of compounds as potential anti-leukemic agents . Treatment of hepatocellular carcinoma with sorafenib - focus on special populations and adverse event management . DB00398 , a receptor tyrosine kinase-inhibitor with anti-proliferative and anti-angiogenic activity , is currently the only approved systemic treatment for patients with hepatocellular carcinoma . It inhibits downstream signaling of P35968 , P09619 , c-Kit receptors and P15056 . Over the last four years comprehensive experience with sorafenib in this indication has been accumulated . In this review we discuss the current data on the use of sorafenib in patients with advanced HCC including special patient populations such as patients with impaired liver function , patients after transplantation , and others . The most frequent side-effects and practical tips on how to manage them are discussed in detail . In addition , we summarize the current experimental data on the use of sorafenib in combination treatment , e. g. , together with transarterial chemoembolisation or other targeted agents . Transduction of P28906 + cells with lentiviral vectors enables the production of large quantities of transgene-expressing immature and mature dendritic cells . BACKGROUND : Genetically engineered dendritic cells ( DC ) presenting specific antigens to T cells may be of great interest for immunotherapy . For this reason , the production of transgene-expressing DC derived from P28906 + cells transduced either shortly after ex vivo purification or during their differentiation into DC were evaluated . METHODS : P28906 + cells were transduced with lentivectors encoding for GFP before or after 21 days of culture with P36888 -ligand , thrombopoietin and stem cell factor and induction into DC with GM- P04141 + P05112 ( G4 ) or G4+ P01375 ( GT4 ) . GFP and DC-specific marker expression was assessed by flow cytometry , and allostimulatory capacity was evaluated on GFP+ and GFP- sorted cells . RESULTS : Immature ( G4-induced ) DC obtained from amplified P28906 + cells were transducible by lentiviral vectors while mature ( GT4-induced ) DC were rather refractory . Moreover , since differentiated DC did not proliferate , large quantities of vectors were required to generate transgene-expressing cells with this protocol . In contrast , greater numbers of both immature and mature GFP- expressing DC were obtained with P28906 + cells exposed to lentivector shortly after purification . By the time of DC induction , GFP+ cells had increased by approximately 170-fold . After DC induction with G4 , 32 % of CD1a+ , HLA-DR+ , or P25942 + cells expressed GFP . CD1a+ P12830 + GFP+ Langerhans-like DC were also obtained . Incubation with P01375 induced mature Q01151 +GFP+ DC that displayed a higher allostimulatory capacity than cells induced with G4 alone . CONCLUSION : The transduction of a small number of P28906 + cells with minimal doses of lentivector may allow for the production of a large number of DC expressing selected antigens useful for immunotherapy . Loss of Jak2 impairs endothelial function by attenuating P04049 / Q02750 /Sp-1 signaling along with altered P29474 activities . A number of inhibitors have been used to dissect the functional relevance of Jak2 in endothelial homeostasis , with disparate results . Given that Jak2 deficiency leads to embryonic lethality , the exact role of Jak2 in the regulation of postnatal endothelial function is yet to be fully elucidated . We generated a model in which Jak2 deficiency can be induced by tamoxifen in adult mice . Loss of Jak2 significantly impaired endothelium-dependent response capacity for vasodilators . Matrigel plug assays indicated a notable decrease in endothelial angiogenic function in Jak2-deficient mice . Studies in a hindlimb ischemic model indicated that Jak2 activity is likely to be a prerequisite for prompt perfusion recovery , based on the concordance of temporal changes in Jak2 expression during the course of ischemic injury and perfusion recovery . A remarkable delay in perfusion recovery , along with reduced capillary and arteriole formation , was observed in Jak2-deficient mice . Antibody array studies indicated that loss of Jak2 led to repressed P29474 expression . In mechanistic studies , Jak2 deficiency attenuated P04049 / Q02750 signaling , which then reduced activity of Sp-1 , an essential transcription factor responsible for P29474 expression . These data are important not only for understanding the exact role that Jak2 plays in endothelial homeostasis , but also for assessing Jak2-based therapeutic strategies in a variety of clinical settings . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . Therapeutic P18621 pathway blockade augments with other modalities of immunotherapy T-cell function to prevent immune decline in ovarian cancer . The tumor microenvironment mediates induction of the immunosuppressive programmed cell death-1 ( P18621 ) pathway , and targeted interventions against this pathway can help restore antitumor immunity . To gain insight into these responses , we studied the interaction between P18621 expressed on T cells and its ligands ( P18621 : Q9NZQ7 , P18621 : Q9BQ51 , and Q9NZQ7 : P33681 .1 ) , expressed on other cells in the tumor microenvironment , using a syngeneic orthotopic mouse model of epithelial ovarian cancer ( ID8 ) . Exhaustion of tumor-infiltrating lymphocytes ( Q15399 ) correlated with expression of P18621 ligands by tumor cells and tumor-derived myeloid cells , including tumor-associated macrophages ( TAM ) , dendritic cells , and myeloid-derived suppressor cells ( MDSC ) . When combined with GVAX or FVAX vaccination ( consisting of irradiated ID8 cells expressing granulocyte macrophage colony-stimulating factor or P36888 ligand ) and costimulation by agonistic α-4-1BB or TLR 9 ligand , antibody-mediated blockade of P18621 or Q9NZQ7 triggered rejection of ID8 tumors in 75 % of tumor-bearing mice . This therapeutic effect was associated with increased proliferation and function of tumor antigen-specific effector CD8(+) T cells , inhibition of suppressive regulatory T cells ( Treg ) and MDSC , upregulation of effector T-cell signaling molecules , and generation of T memory precursor cells . Overall , P18621 / Q9NZQ7 blockade enhanced the amplitude of tumor immunity by reprogramming suppressive and stimulatory signals that yielded more powerful cancer control . Sensitivity toward sorafenib and sunitinib varies between different activating and drug-resistant P36888 -ITD mutations . OBJECTIVE : Activating mutations in P36888 are known to be a frequent transforming event in acute myeloid leukemia . Small molecule-inhibitor therapy targeting the P36888 kinase is , therefore , an attractive strategy . P36888 kinase inhibitors , such as PKC412 , have already entered clinical trials . Even though results are encouraging , emergence of primary and secondary resistance does occur in the majority of patients . Thus , it will be crucial to carefully characterize the activity of every single compound against different activating and resistance P36888 -internal tandem duplication ( ITD ) mutations . Here we tested the efficacy of sunitinib and sorafenib to inhibit primary P36888 activating mutations ( ITD and D835Y ) and of secondary resistance mutations . METHODS : Ba/ P13726 cell lines stably expressing oncogenic P36888 mutations were used to calculate cellular IC(50) values for sunitinib and sorafenib using cell proliferation assays . Differential IC(50) values for sorafenib toward P36888 -ITD and P36888 -D835Y were confirmed by Western blotting . Cell death was measured by propidium-iodide staining and flow cytometry . RESULTS : DB00398 inhibits P36888 -ITD more potent than P36888 -D835Y , while sunitinib is equally effective against both mutant forms of P36888 . Importantly , sensitivity toward sorafenib and sunitinib varies between the different secondary P36888 -ITD resistance mutations . CONCLUSIONS : These results establish sensitivity profiles for the P36888 inhibitors sunitinib and sorafenib . This may help to develop rational treatment strategies for acute myeloid leukemia with these compounds . Sustained response following sorafenib therapy in an older adult patient with advanced renal cancer on hemodialysis : a case report . The prognosis for patients with renal cell carcinoma is very poor , with a five-year survival rate of less than 10 % . DB00398 is an orally administered multikinase inhibitor that blocks intracellular kinases in the Raf/MEK/ P29323 pathway involved in tumor proliferation , and also kinases responsible for angiogenesis , including VEGFr-2 , VEGFr-3 , Flt-3 , PDGFr-β and c- P10721 . As a consequence of its limited renal clearance , sorafenib appears to be suitable for patients with advanced kidney cancer and terminal renal failure . The case of a 72-year-old male patient on hemodialysis and receiving sorafenib treatment for mRCC is reported . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . VIGS approach reveals the modulation of anthocyanin biosynthetic genes by CaMYB in chili pepper leaves . The purple coloration of pepper leaves arises from the accumulation of anthocyanin . Three regulatory and 12 structural genes have been characterized for their involvement in the anthocyanin biosynthesis . Examination of the abundance of these genes in leaves showed that the majority of them differed between anthocyanin pigmented line Z1 and non-pigmented line A3 . Silencing of the R2R3- P10242 transcription factor CaMYB in pepper leaves of Z1 resulted in the loss of anthocyanin accumulation . Moreover , the expression of multiple genes was altered in the silenced leaves . The expression of MYC was significantly lower in CaMYB-silenced leaves , whereas WD40 showed the opposite pattern . Most structural genes including Q99698 , CHI , F3H , P13726 '5'H , DFR , ANS , UFGT , P01160 , and Q86UG4 were repressed in CaMYB-silenced foliage with the exception of Q9P2V4 , C4H , and 4CL . These results indicated that P10242 plays an important role in the regulation of anthocyanin biosynthetic related genes . Besides CaMYB silenced leaves rendered more sporulation of Phytophthora capsici Leonian indicating that CaMYB might be involved in the defense response to pathogens . Toxicity of sorafenib : clinical and molecular aspects . IMPORTANCE OF THE FIELD : DB00398 is a novel oral bis-aryl urea compound originally developed as an inhibitor of RAF kinase for its anti-proliferative property . DB00398 also inhibits receptor tyrosine kinases of multiple pro-angiogenic factors such as P17948 /2/3 , Flt-3 and P09619 . The combination of both its anti-proliferative and anti-angiogenic properties makes sorafenib an attractive agent in cancer treatment . DB00398 has been approved for the treatment of metastatic renal cell carcinoma as well as hepatocellular cancer . Despite its inherent selectivity , sorafenib can cause unusual adverse events whose the management represents a challenge for oncologists . AREAS COVERED IN THIS REVIEW : Relevant literature was identified using a Pubmed search of articles published up to June 2009 . Search terms included ' sorafenib ' and ' toxicity ' . Original articles were reviewed and relevant citations from these articles were also considered . WHAT THE READER WILL GAIN : The clinical aspect of sorafenib-induced adverse events and the molecular basis behind this toxicity are discussed . Finally , recommendations for the management of these adverse events are proposed . TAKE HOME MESSAGE : Although not life-threatening , toxicity of sorafenib can severely impact the physical , psychological and social well-being of patients . The management of this unusual toxicity highlights the particular need of new pluridisciplinarities linking oncologist , cardiologist and dermatologist . Phase II study of sorafenib in children with recurrent or progressive low-grade astrocytomas . BACKGROUND : Activation of the DB01367 -RAF-MEK- P29323 signaling pathway is thought to be the key driver of pediatric low-grade astrocytoma ( PLGA ) growth . DB00398 is a multikinase inhibitor targeting P15056 , VEGFR , P09619 , and c-kit . This multicenter phase II study was conducted to determine the response rate to sorafenib in patients with recurrent or progressive PLGA . METHODS : Key eligibility criteria included age ≥ 2 years , progressive PLGA evaluable on Q9BWK5 , and at least one prior chemotherapy treatment . DB00398 was administered twice daily at 200 mg/m(2)/dose ( maximum of 400 mg/dose ) in continuous 28-day cycles . Q9BWK5 , including 3-dimensional volumetric tumor analysis , was performed every 12 weeks . P15056 molecular testing was performed on tumor tissue when available . RESULTS : Eleven patients , including 3 with neurofibromatosis type 1 ( P21359 ) , were evaluable for response ; 5 tested positive for P15056 duplication . Nine patients ( 82 % ) came off trial due to radiological tumor progression after 2 or 3 cycles , including 3 patients with confirmed P15056 duplication . Median time to progression was 2.8 months ( 95 % CI , 2.1-31.0 months ) . Enrollment was terminated early due to this rapid and unexpectedly high progression rate . Tumor tissue obtained from 4 patients after termination of the study showed viable pilocytic or pilomyxoid astrocytoma . CONCLUSIONS : DB00398 produced unexpected and unprecedented acceleration of tumor growth in children with PLGA , irrespective of P21359 or tumor P15056 status . In vitro studies with sorafenib indicate that this effect is likely related to paradoxical P29323 activation . Close monitoring for early tumor progression should be included in trials of novel agents that modulate signal transduction . Phase I study of the combination of sorafenib and temsirolimus in patients with metastatic melanoma . PURPOSE : This phase I clinical trial was conducted to determine the safety , efficacy , and molecular effects of sorafenib with temsirolimus in patients with advanced melanoma . PATIENTS AND METHODS : Patients with stage IV or unresectable or recurrent stage III melanoma and Eastern Cooperative Oncology Group performance status of 0 to 1 were eligible . DB00398 was given orally once or twice daily and temsirolimus was given i.v. weekly , both starting on day 1 , with a 4-week cycle . Responses were assessed every 2 cycles per Response Evaluation Criteria in Solid Tumors criteria . Consenting patients with accessible tumors underwent optional tumor biopsies before treatment and after the second infusion of temsirolimus . Tumor biopsies were analyzed for activating mutations in P15056 and P01111 , and for expression of P-extracellular signal-regulated kinase ( P- P29323 ) and P-S6 proteins . RESULTS : A total of 25 patients were accrued to the study . The maximum tolerated doses were sorafenib 400 mg every morning and 200 mg every evening and temsirolimus 25 mg i.v. weekly . Dose-limiting toxicities included thrombocytopenia , hand-foot syndrome , serum transaminase elevation , and hypertriglyceridemia . There were no complete or partial responses with the combination ; 10 patients achieved stabilization of disease as their best response . The median progression-free survival was 2.1 months . Matching pretreatment and day 15 tumor biopsies showed marked inhibition of P-S6 with treatment in 3 of 4 evaluable patients , but minimal inhibition of P- P29323 . CONCLUSIONS : Combination therapy with sorafenib and temsirolimus resulted in significant toxicity at higher dose levels , failed to achieve any clinical responses in genetically unselected patient population , and did not inhibit P- P29323 . Bypassing tumor-specific and bispecific antibodies : triggering of antitumor immunity by expression of anti-FcgammaR scFv on cancer cell surface . We have developed a novel immunostimulatory molecule against tumor cells , composed of an anti-FcgammaRIII ( CD16 ) scFv fused to the platelet-derived growth factor receptor ( P09619 ) transmembrane region . This fusion molecule was stably expressed on the tumor cell surface and retained the ability of the parental antibody to bind soluble CD16 . Tumor cells expressing anti-CD16 scFv triggered the release of P60568 by Jurkat-CD 16/gamma cells and of TNFalpha by monocytes when co-cultured with these cells . Furthermore , NK cells could kill scFv-transfected HLA+ class I H1299 lung carcinoma tumor cells , but not the parental cells , indicating that anti-CD16 scFv tumor expression prevents the killer inhibitory receptor ( P55040 ) -mediated inhibition of NK cell cytotoxicity . This anti-CD16 scFv tumor expression also enhanced tumor phagocytosis by IFNgamma-activated macrophages , a mechanism known to induce a protective long-term adaptative immunity to tumors . In vivo Winn tests performed in SCID mice showed that the expression of anti-CD16 scFv on tumor cells , but not of the negative control anti-phOx scFv , prevented tumor cell growth . Thus , expression of FcR antibodies or other FcR-specific ligands on tumor cells represents a novel and potent antibody-based gene therapy approach , which may have clinical applications in cancer Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Peripheral blood gene expression of P33681 and P10747 family members associated with tumor progression and microscopic lymphovascular invasion in colon cancer patients . PURPOSE : To associate the global gene expression of P33681 / P10747 family transcripts with pathologic features of colon cancer , we determined the P33681 / P10747 family transcripts in peripheral blood mononuclear cells ( PBMCs ) from normal subjects and patients with adenomatous polyps and colon cancer , and correlated the results with pathologic features of colon cancer . METHODS : PBMCs from age-matched normal subjects and patients with adenomatous polyps and colon cancer were analyzed for peripheral blood transcripts ( PBTs ) of P33681 / P10747 family using real-time PCR . Differences in expression levels of P33681 / P10747 PBTs across all cancer stages and between colon cancer patients with or without microscopic lymphovascular invasion ( LVI ) were analyzed . RESULTS : The results showed a significant upregulation of PBTs of co-inhibitory molecules such as Q5ZPR3 and P18621 and a significant P10721 downregulation of co-stimulatory molecules including P10747 and Q9Y6W8 in colon cancer patients . Furthermore , the increase of Q5ZPR3 P10721 was strongly associated with tumor invasion ( P = 0.025 ) and advanced TNM stages ( P = 0.019 ) , whereas the decline of co-stimulatory ligand O75144 P10721 was related to regional lymph node metastasis ( P = 0.028 ) and aggressive tumor invasion ( P = 0.031 ) . In addition , the ratios of P10721 expression of P10747 family to P33681 family such as P16410 to O75144 and P18621 to O75144 were significantly higher in colon cancer patients with microscopic LVI than in those without LVI ( P = 0.001 and P = 0.016 , respectively ) . CONCLUSIONS : Our results suggest that P33681 / P10747 family PBTs may serve as valuable markers reflecting the pathological features of colon cancer . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . A randomized phase I clinical and biologic study of two schedules of sorafenib in patients with myelodysplastic syndrome or acute myeloid leukemia : a NCIC ( National Cancer Institute of Canada ) Clinical Trials Group Study . DB00398 is a small molecule inhibitor of RAF kinase , P35968 , c- P10721 , and P36888 . In this randomized phase I study , eligible patients had relapsed/refractory acute myeloid leukemia ( AML ) , and one prior induction regimen , or were age > 65 with untreated myelodysplastic syndrome ( P43034 ) or secondary AML . DB00398 was given orally for 28 days ( cont ) or 14 days ( int ) every 4 weeks at three dose levels ( 100 , 200 , and 400 mg P55957 ) ; 300 mg cont was also tested . Forty-two patients were enrolled ( median age 71 [ 37-82 ] ; prior chemotherapy : 22 ) . Dose-limiting toxicity ( DLT ) was : 100 mg P55957 : 0/7 patients ; 200 mg P55957 : 2/12 patients ; 400 mg P55957 : 1/17 patients . DB00398 400 mg cont was not tolerated in this population : 6/8 received < 14 days of treatment due to toxicity ; no DLT was seen with 300 mg cont . One CR was seen in a patient with AML with P36888 -ITD . Flow cytometry studies suggest that sorafenib inhibits P29323 phosphorylation via c- P10721 . The recommended phase II dose in AML is 300 mg P55957 continuously , and testing in combination and in P36888 -ITD AML is warranted .

MH_train_1097

MH_train_1097

MH_train_1097

interacts_with DB08918?

multiple_choice

An aging pathway controls the TrkA to p75NTR receptor switch and amyloid beta-peptide generation . Aging of the brain is characterized by marked changes in the expression levels of the neurotrophin receptors , TrkA and p75(NTR) . An expression pattern in which TrkA predominates in younger animals switches to one in which p75(NTR) predominates in older animals . This TrkA-to-p75(NTR) switch is accompanied by activation of the second messenger ceramide , stabilization of beta-site amyloid precursor protein-cleaving enzyme-1 ( P56817 ) , and increased production of amyloid beta-peptide ( Abeta ) . Here , we show that the insulin-like growth factor-1 receptor ( IGF1-R ) , the common regulator of lifespan and age-related events in many different organisms , is responsible for the TrkA-to-p75(NTR) switch in both human neuroblastoma cell lines and primary neurons from mouse brain . The signaling pathway that controls the level of TrkA and p75(NTR) downstream of the IGF1-R requires Q9Y4H2 , PIP3/Akt , and is under the control of P60484 and Q8TCB0 , the short isoform of p53 . We also show that hyperactivation of IGF1-R signaling in Q8TCB0 transgenic animals , which show an accelerated form of aging , is characterized by early TrkA-to-p75(NTR) switch and increased production of Abeta in the brain . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . Genetic pathway-based hierarchical clustering analysis of older adults with cognitive complaints and amnestic mild cognitive impairment using clinical and neuroimaging phenotypes . Hierarchical clustering is frequently used for grouping results in expression or haplotype analyses . These methods can elucidate patterns between measures that can then be applied to discerning their validity in discriminating between experimental conditions . Here a hierarchical clustering method is used to analyze the results of an imaging genetics study using multiple brain morphology and cognitive testing endpoints for older adults with amnestic mild cognitive impairment ( D6RGH6 ) or cognitive complaints ( CC ) compared to healthy controls ( HC ) . The single nucleotide polymorphisms ( SNPs ) are a subset of those included on a larger array that are found in a reported Alzheimer 's disease ( AD ) and neurodegeneration pathway . The results indicate that genetic models within the endpoints cluster together , while there are 4 distinct sets of SNPs that differentiate between the endpoints , with most significant results associated with morphology endpoints rather than cognitive testing of patients ' reported symptoms . The genes found in at least one cluster are P08183 , Q02410 , P56817 , Q9Y5Z0 , P10415 , Q07817 , P55210 , P28329 , P01034 , P35462 , P21918 , P05231 , Q07954 , NAT1 , and P49810 . The greater associations with morphology endpoints suggests that changes in brain structure can be influenced by an individual 's genetic background in the absence of dementia and in some cases ( Cognitive Complaints group ) even without those effects necessarily being detectable on commonly used clinical tests of cognition . The results are consistent with polygenic influences on early neurodegenerative changes and demonstrate the effectiveness of hierarchical clustering in identifying genetic associations among multiple related phenotypic endpoints . Molecular determinants of trastuzumab efficacy : What is their clinical relevance ? DB00072 -containing therapy is a standard of care for human epidermal growth factor receptor-2 ( P04626 ) -positive breast cancer . In pre-clinical models , a wide range of molecular mechanisms have been associated with reduced sensitivity to trastuzumab in vitro . These include expression of the truncated P04626 receptor fragment p95HER2 , activating mutation of the gene encoding the class 1A catalytic subunit of phosphatidylinositol 3-kinase ( P42336 ) , loss of phosphatase and tensin homolog ( P60484 ) , activation of other downstream signal transducers , prevention of cell cycle arrest , increased signaling through alternative ( HER or non-HER ) tyrosine kinase receptors , and resistance to antibody-dependent cellular cytotoxicity . However , the clinical significance of these mechanisms as determinants of trastuzumab efficacy in vivo has been unclear . Here , we review clinical studies of potential predictive biomarkers of trastuzumab efficacy in P04626 -positive breast cancer and consider whether evaluation of such markers might inform patient selection for therapy . We find that clinical evidence relating to potential predictive biomarkers is mostly limited to small , retrospective studies , many of which have yielded conflicting findings . Some trends are evident in the retrospective data and in biomarker analyses from randomized clinical trials , particularly relating to activation of the phosphatidylinositol 3-kinase pathway , but none is sufficiently strong to form a basis for patient selection . This may be explained by the fact that multiple mechanisms of action determine the clinical efficacy of trastuzumab . In the absence of novel , validated biomarkers of efficacy , trastuzumab eligibility should continue to be based on evaluation of P04626 status according to standard methods . Transcriptional regulation of beta-secretase by p25/cdk5 leads to enhanced amyloidogenic processing . P12004 -dependent kinase 5 ( cdk5 ) has been implicated in Alzheimer 's disease ( AD ) pathogenesis . Here , we demonstrate that overexpression of p25 , an activator of cdk5 , led to increased levels of P56817 mRNA and protein in vitro and in vivo . A p25/cdk5 responsive region containing multiple sites for signal transducer and activator of transcription ( P42224 /3 ) was identified in the P56817 promoter . P40763 interacts with the P56817 promoter , and p25-overexpressing mice had elevated levels of pSTAT3 and P56817 , whereas cdk5-deficient mice had reduced levels . Furthermore , mice with a targeted mutation in the P40763 cdk5 responsive site had lower levels of P56817 . Increased P56817 levels in p25 overexpressing mice correlated with enhanced amyloidogenic processing that could be reversed by a cdk5 inhibitor . These data demonstrate a pathway by which p25/cdk5 increases the amyloidogenic processing of P05067 through P40763 -mediated transcriptional control of P56817 that could have implications for AD pathogenesis . Granulocyte macrophage-colony stimulating factor increases the expression of histamine and histamine receptors in monocytes/macrophages in relation to arteriosclerosis . OBJECTIVE : To study the effect of granulocyte macrophage-colony-stimulating factor ( GM- P04141 ) on histamine metabolism in arteriosclerosis , the expression of histidine decarboxylase ( HDC ; histamine-producing enzyme ) , histamine receptors 1 and 2 ( P35367 and P25021 ) , and GM- P04141 was investigated in human and mouse arteriosclerotic carotid arteries . Furthermore , the molecular mechanisms of GM- P04141 -induced HDC and P35367 expression in monocytic U937 cells were investigated . METHODS AND RESULTS : Immunohistochemistry showed that atherosclerotic human coronary and mouse ligated carotid arteries contained HDC-expressing macrophages . Gene expression of HDC , P35367 , P25021 , and GM- P04141 was also detected in the lesions . In U937 cells , GM- P04141 enhanced histamine secretion and gene expression of HDC and P35367 . A promoter assay showed that GM- P04141 enhanced gene transcription of HDC and P35367 but not P25021 . CONCLUSIONS : The present results indicate that HDC and HHR are expressed in arteriosclerotic lesion , and that GM- P04141 induces HDC and P35367 expression in monocytes . Locally produced histamine might participate in atherogenesis by affecting the expression of atherosclerosis-related genes in monocytes and smooth muscle cells . The presence of histamine-producing macrophages and gene expression of histamine receptors and GM- P04141 was demonstrated in arteriosclerotic lesions . In monocytic U937 cells , GM- P04141 upregulated the expression of histamine and P35367 . Coordinated expression of histamine and its receptors by GM- P04141 would participate in atherogenesis by affecting monocytic and SMC gene expression . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Deletion of the prostaglandin E2 EP2 receptor reduces oxidative damage and amyloid burden in a model of Alzheimer 's disease . Epidemiological studies demonstrate that chronic use of nonsteroidal anti-inflammatory drugs ( NSAIDs ) in normal aging populations reduces the risk of developing Alzheimer 's disease ( AD ) . NSAIDs inhibit the enzymatic activity of cyclooxygenase-1 ( P23219 ) and inducible P35354 , which catalyze the first committed step in the synthesis of prostaglandins . These studies implicate P36551 -mediated inflammation as an early and potentially reversible preclinical event ; however , the mechanism by which P36551 activity promotes development of AD has not been determined . Recent studies implicate the prostaglandin E2 ( DB00917 ) E prostanoid subtype 2 ( EP2 ) receptor in the development of the innate immune response in brain . Here , we report that deletion of the DB00917 EP2 receptor in the APPSwe-PS1DeltaE9 model of familial AD results in marked reductions in lipid peroxidation in aging mice . This reduction in oxidative stress is associated with significant decreases in levels of amyloid-beta ( Abeta ) 40 and 42 peptides and amyloid deposition . Aged APPSwe-PS1DeltaE9 mice lacking the EP2 receptor harbor lower levels of beta C-terminal fragments , the product of beta-site P05067 cleaving enzyme ( P56817 ) processing of amyloid precursor protein . Increases in P56817 processing have been demonstrated in models of aging and AD and after oxidative stress . Our results indicate that DB00917 signaling via the EP2 receptor promotes age-dependent oxidative damage and increased Abeta peptide burden in this model of AD , possibly via effects on P56817 activity . Our findings identify EP2 receptor signaling as a novel proinflammatory and proamyloidogenic pathway in this model of AD , and suggest a rationale for development of therapeutics targeting the EP2 receptor in neuroinflammatory diseases such as AD . Combined treatment with a P56817 inhibitor and anti-Aβ antibody gantenerumab enhances amyloid reduction in APPLondon mice . Therapeutic approaches for prevention or reduction of amyloidosis are currently a main objective in basic and clinical research on Alzheimer 's disease . Among the agents explored in clinical trials are anti-Aβ peptide antibodies and secretase inhibitors . Most anti-Aβ antibodies are considered to act via inhibition of amyloidosis and enhanced clearance of existing amyloid , although secretase inhibitors reduce the de novo production of Aβ . Limited information is currently available on the efficacy and potential advantages of combinatorial antiamyloid treatment . We performed a chronic study in APPLondon transgenic mice that received treatment with anti-Aβ antibody gantenerumab and P56817 inhibitor RO5508887 , either as mono- or combination treatment . Treatment aimed to evaluate efficacy on amyloid progression , similar to preexisting amyloidosis as present in Alzheimer 's disease patients . Mono-treatments with either compound caused a dose-dependent reduction of total brain Aβ and amyloid burden . Combination treatment with both compounds significantly enhanced the antiamyloid effect . The observed combination effect was most pronounced for lowering of amyloid plaque load and plaque number , which suggests effective inhibition of de novo plaque formation . Moreover , significantly enhanced clearance of pre-existing amyloid plaques was observed when gantenerumab was coadministered with RO5508887 . P56817 inhibition led to a significant time- and dose-dependent decrease in P04141 Aβ , which was not observed for gantenerumab treatment . Our results demonstrate that combining these two antiamyloid agents enhances overall efficacy and suggests that combination treatments may be of clinical relevance . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 .

MH_train_1098

MH_train_1098

MH_train_1098

interacts_with DB00328?

multiple_choice

Increased epithelial permeability is the primary cause for bicarbonate loss in inflamed murine colon . BACKGROUND : Bicarbonate loss into the lumen occurs during intestinal inflammation in different species . However , candidate pathways like P13569 or P40879 are inhibited in the inflamed gut . This study addressed the question whether and how inflammation-associated increased intestinal permeability may result in epithelial HCO(3)(-) loss . METHODS : Murine proximal colon was studied because it does not express functional P40879 but is inflamed in the tumor necrosis factor α overexpressing mouse model ( P01375 (ΔARE) ) . DB01174 alkalization , (3)H-mannitol fluxes , impedance spectroscopy , and dilution potentials were measured in Ussing chambers , whereas expression and localization of tight junction-associated proteins were analyzed by Western blots and immunohistochemistry . RESULTS : DB01174 alkalization rates and (3)H-mannitol fluxes were increased in P01375 (+/ΔARE) proximal colon , whereas forskolin-stimulated I(sc) was not altered . Epithelial resistance was reduced , but subepithelial resistance increased . The epithelial lining was intact , and enterocyte apoptosis rate was not increased despite massively increased Th1 cytokine levels and lymphoplasmacellular infiltration . Measurement of dilution potentials suggested a loss of cation selectivity with increased anion permeability . Western analysis revealed a downregulation of occludin expression and an upregulation of both claudin-2 and claudin-5 , with no change in ZO-1 , P12830 , claudin-4 , and claudin-8 . Immunohistochemistry suggested correct occludin localization but reduced tight junction density in P01375 (+/ΔARE) surface epithelium . CONCLUSIONS : Inflammation during P01375 -α overexpression leads to increased epithelial permeability in murine proximal colon , decreased tight junctional cation selectivity , and increased HCO(3)(-) loss into the lumen . Inflammation-associated colonic HCO(3)(-) loss may occur through leaky tight junctions rather than through HCO(3)(-) secreting ion transporters . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . A 3-D model for P08908 -receptor agonists based on stereoselective methyl-substituted and conformationally restricted analogues of 8-hydroxy-2-(dipropylamino)tetralin . The enantiomers of cis- and trans-1,2,3,4,4a,5,10,10a-octahydro-9-hydroxy-1- propylbenzo[g]quinolines ( 10 and 11 , respectively ) and the enantiomers of trans-1,2,3,4,4a,5,6,10b-octahydro-10- hydroxy-4-propylbenzo[f]quinoline ( 12 ) have been synthesized and their stereochemical and conformational characteristics have been studied by use of X-ray crystallography and molecular mechanics ( P08253 ) calculations . The compounds , which are conformationally restricted analogues of the potent 5-hydroxytryptamine ( 5-HT ) receptor agonist 8-hydroxy-2- (dipropylamino)tetralin ( 8-OH-DPAT ; 1 ) have been evaluated for central 5-HT and dopamine receptor stimulating activity by use of biochemical and behavioral tests in rats . In addition , we have evaluated the ability of these compounds and a number of previously reported analogues to displace [ 3H ] -8-OH-DPAT from P08908 -binding sites . The enantiomers of 12 behave as potent P08908 -receptor agonists , whereas the octahydrobenzo[g]quinoline derivatives are much less potent or inactive . In general , the affinities of the compounds correlate well with their agonist potencies . The set of compounds under study is accommodated by a novel computer-graphics-derived model for P08908 -receptor agonism . The model consists of a flexible pharmacophore and a partial receptor-excluded volume . Identification of the amino acid sequence motif of alpha-synuclein responsible for macrophage activation . P37840 ( Syn ) is implicated in the pathogenesis of PD and related neurodegenerative disorders . Recent studies have also shown that alpha-synuclein can activate microglia and enhance dopaminergic neurodegeneration . The mechanisms of microglia activation by alpha-synuclein , however , are not well understood . In this study , we found that not only alpha-synuclein but also beta- and gamma-synucleins activated macrophages ( RAW 264.7 ) in vitro . Macrophages treated with synuclein proteins secreted P01375 in a dose-dependent manner . Synuclein family proteins also increased mRNA transcription of P35354 and P35228 . Two alpha-synuclein deletion mutants , SynDeltaNAC and Syn61-140 , activated macrophages , while deletion mutants Syn1-60 and Syn96-140 did not significantly activate them . Finally , we demonstrated that macrophage activation by alpha-synuclein was accompanied by phosphorylation of P29323 . These results suggest that synuclein family proteins can activate macrophages , and that macrophage activation needs both the N-terminal and C-terminal domains of alpha-synuclein , but not the central Q9C000 region . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity . Evaluation of pharmacological profile of meloxicam as an anti-inflammatory agent , with particular reference to its relative selectivity for cyclooxygenase-2 over cyclooxygenase-1 . We studied the anti-inflammatory activity of meloxicam on rat carrageenin-induced pleurisy and its toxicity for rat gastric mucosa , relative to its in vitro inhibitory potency against partially purified cyclooxygenase ( P36551 ) -1 and P35354 preparations in order to clarify the pharmacological profile of the compound as an anti-inflammatory agent . In rat carrageenin-induced pleurisy , the plasma exudation rate peaked at 5 h , at which time P35354 was detectable in cells from the pleural exudate . Meloxicam and piroxicam ( 1 and 3 mg/kg ) and NS-398 ( 3 mg/kg ) showed almost equal anti-inflammatory potency against 5-hour pleurisy . A single oral administration of the compounds caused a dose-dependent increase in the number of rats with gastric mucosal erosion . The ED50 value for meloxicam ( 5.92 mg/kg ) was significantly higher than that for piroxicam ( 1.76 mg/kg ) , indicating that meloxicam is safer . DB00328 showed intermediate safety ( 2.59 mg/kg ) . In in vitro experiments , indometacin inhibited P23219 about 1.7 times more potently than P35354 . NS-398 inhibited P35354 with an IC50 of 0.32 microM , but never affected P23219 activity , even at 100 microM . In the same assay system , meloxicam inhibited P35354 about 12 times more selectively than P23219 . Piroxicam , however , inhibited both isoforms almost equally . These results indicate that meloxicam is a potent anti-inflammatory agent with low gastric toxicity . One reason for its in vivo pharmacological profile may be related to its relative selectivity for P35354 over P23219 . Thus , meloxicam may belong to a group of P35354 selective anti-inflammatory agents with a better safety profile than conventional P23219 and P35354 nonselective anti-inflammatory agents . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB00328 ameliorates high glucose-induced proliferation and invasion via modulation of e-cadherin in pancreatic cancer cells . DB00328 , an inhibitor of cyclooxygenase-2 ( P35354 ) , has been shown to exert anticancer effects in a variety of cancers . However , the effect and mechanism of indometacin on high glucose ( HG ) -induced proliferation and invasion of pancreatic cancer ( PC ) cells remain unclear . Multiple lines of evidence suggest that a large portion of pancreatic cancer ( PC ) patients suffer from either diabetes or HG which contributing PC progression . In this study , we report that indometacin down-regulated HG-induced proliferation and invasion via up-regulating P12830 but not P35354 in PC cells . Additionally , the P12830 transcriptional repressors , Snail and Slug , were also involved in the process . Furthermore , the proliferation and invasion of PC cells , incubated in HG medium and treated with indometacin were significantly increased when P12830 was knocked down ( Si-E-cad ) . Moreover , the protein levels of P08253 , P14780 , and P15692 were increased in PC cells transfected with Si-E-cad . Finally , the activation of the PI3K/AKT/GSK-3β signaling pathway was demonstrated to be involved in indometacin reversing HG-induced cell proliferation and invasion in PC cells . In conclusion , these results suggest that indometacin plays a key role in down-regulating HG-induced proliferation and invasion in PC cells . Our findings indicate that indometacin could be used as a novel therapeutic strategy to treat PC patients who simultaneously suffer from diabetes or HG . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . The protective effect of rebamipide on paracellular permeability of rat gastric epithelial cells . BACKGROUND : Barrier function in gastric epithelial cells is essential for the gastric defence mechanism against acid back-diffusion into the mucosal layer . Our previous study indicated that trans-epithelial resistance ( Q9NZ01 ) of rat gastric epithelial cells was rapidly increased when the cells were exposed to acid . This response to acid was diminished by indometacin . AIM : Evaluate the effects of a mucoprotective agent , rebamipide , on the nonsteroidal anti-inflammatory drug ( NSAID ) -induced increase of gastric epithelial permeability . METHODS : Rat gastric epithelial cells were plated on tissue culture inserts . Cells were exposed to a NSAID ( indometacin , 10-7 M ) . Trans-epithelial permeability was measured by Q9NZ01 and diffusion rate of 14C-mannitol . The effect of rebamipide was evaluated by measuring Q9NZ01 . Endogenous prostaglandin E2 ( DB00917 ) production in culture medium was also measured . RESULTS : DB00328 gradually and significantly decreased Q9NZ01 and increased 14C-manitol permeability . Rebamipide reversed the indometacin-induced changes in epithelial permeability and induced DB00917 synthesis . This induction was blocked by either indometacin or a Cyclooxygenase ( P36551 ) -2 specific inhibitor . CONCLUSIONS : P36551 inhibitors such as indometacin inhibit regulation of epithelial permeability by reducing DB00917 . P23219 has an important role in the gastric defense mechanism . Rebamipide suppressed an indometacin-induced increase in gastric epithelial permeability by increasing DB00917 levels in a P35354 dependent manner . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . The transcriptional coactivator DRIP/mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) /mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 /MED1 ( MED1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED1/ Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and/or glioma-associated oncogene homolog . Blocking MED1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation . P01308 like growth factor-1 selectively regulates the expression of matrix metalloproteinase-2 in malignant H-ras transformed cells . The present study demonstrates alterations in the regulation of matrix metalloproteinase-2 ( P08253 ) expression in response to insulin like growth factor-1 ( DB01277 ) in a H-ras transformed cell line , P01024 , which is capable of metastasis formation . These changes in P08253 expression in response to DB01277 treatment did not occur in either non-transformed parental 10 T 1/2 cells or in H-ras transformed cells ( Q13224 cells ) which are capable of benign tumour formation . DB01277 treatment of P01024 cells resulted in increased expression of P08253 gelatinolytic activity and increased expression of P08253 mRNA levels . The DB01277 mediated alterations in P08253 mRNA levels were dependent upon de novo protein synthesis and independent of transcriptional events , but dependent upon post-transcriptional regulatory events . Most notably , DB01277 can regulate P08253 mRNA expression in P01024 cells through a mechanism involving P08253 message stabilization . This study demonstrates aspects of the temporal regulatory mechanisms of P08253 expression in response to insulin-like growth factor-1 in a H-ras transformed fibrosarcoma cell line capable of metastasis formation and thereby , provides further insight into the altered growth regulatory program associated with H-ras mediated cellular transformation and malignant progression . Targetting esophageal and gastric cancers with monoclonal antibodies . Target therapies and notably monoclonal antibodies are currently being considered for esophageal , gastric , and gastroesophageal junction cancers . P00533 was found to be overexpressed in 60-86 % of gastric or gastroesophageal tumors and in 50-70 % of esophageal cancers . Cetuximab was shown to be a radiosensitizing agent in the treatment of ENT neoplasia . These results led to several phase II encouraging therapeutic trials evaluating the combination of cetuximab with radiochemotherapy in locally advanced esophageal cancers . Numerous encouraging phase II trials evaluating cetuximab combined with chemotherapy in patients with gastric adenocarcinoma or gastroesophageal junction cancer were reported . These promising results are still to be confirmed by the ongoing phase III trials . Several studies reported P04626 overexpression in gastric cancer ( 7-34 % ) , which appeared to be associated with poorer prognosis . DB00072 is a monoclonal antibody directed against the extracellular P04626 domain . The international phase III trial known as ToGA ( DB00072 for Gastric Cancer ) aimed to determine the clinical efficacy and acceptable toxicity profile of trastuzumab in combination with first-line chemotherapy in P04626 -overexpressing gastric or gastroesophageal cancer . Angiogenesis is an essential step in the initial phase of tumorigenesis , and it is normally absent from healthy tissues except for particular physiological situations , such as wound healing . P15692 plays a role in endothelial growth and angiogenesis . DB00112 , a humanized monoclonal anti- P15692 antibody , is currently being studied for gastric cancer . The phase III AVAGAST study , evaluating bevacizumab in association with chemotherapy in advanced gastric adenocarcinoma , did not achieve its primary aim of improved OS in bevacizumab-treated patients . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . P37840 activates microglia by inducing the expressions of matrix metalloproteinases and the subsequent activation of protease-activated receptor-1 . The mutation or overexpression of alpha-synuclein protein plays a pivotal role in the pathogenesis of Parkinson 's disease . In our preliminary experiments , we found that alpha-synuclein induced the expression of matrix metalloproteinases ( MMPs ) ( P03956 , -3 , -8 , and -9 ) in rat primary cultured microglia . Thus , the current study was undertaken to determine the roles of MMPs in alpha-synuclein-induced microglial activation . The inhibition of P08254 , -8 , or -9 significantly reduced NO and reactive oxygen species levels and suppressed the expression of P01375 and IL-1beta . Notably , P22894 inhibitor suppressed P01375 production more efficaciously than P08254 or P14780 inhibitors . Inhibition of P08254 or -9 also suppressed the activities of MAPK , NF-kappaB , and AP-1 . Previously , protease-activated receptor-1 ( P25116 ) has been associated with the actions of MMPs , and thus , we further investigated the role of P25116 in alpha-synuclein-induced inflammatory reactions . A P25116 -specific inhibitor and a P25116 antagonist significantly suppressed cytokine levels , and NO and reactive oxygen species production in alpha-synuclein-treated microglia . Subsequent P25116 cleavage assay revealed that P08254 , -8 , and -9 , but not alpha-synuclein , cleaved the synthetic peptide containing conventional P25116 cleavage sites . These results suggest that MMPs secreted by alpha-synuclein-stimulated microglia activate P25116 and amplify microglial inflammatory signals in an autocrine or paracrine manner . Furthermore , our findings suggest that modulation of the activities of MMPs and/or P25116 may provide a new therapeutic strategy for Parkinson 's disease . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Effects of P04626 amplicon size and genomic alterations of chromosomes 1 , 3 , and 10 on patient response to trastuzumab in metastatic breast cancer . DB00072 is widely used for advanced breast cancer patients with P04626 -amplified tumors . Nevertheless , over half of these patients do not have an objective response . One reason may be altered expression of genes that might compensate for P04626 inhibition . We previously mapped the gene-rich region of chromosome 17 telomeric to P04626 , and reported considerable variability in the telomeric extent of the P04626 amplicon . Here we examined whether the variable amplicon size may be associated with patient response to trastuzumab . In addition , we looked at associations between response and several signaling pathway-related genes unrelated to the P04626 amplicon , including Q9Y243 , P60484 , P42336 , and P35354 . In 35 patients with P04626 -amplified metastatic breast cancer , with 40 % overall response to trastuzumab , fluorescence in situ hybridization identified the telomeric extent of the P04626 amplicon and the status of the several pathway-related genes . Objective response strongly correlated with the telomeric amplicon size , with 62 % of patients with shorter amplicons responding , compared with only 7 % of patients with longer amplicons ( P = 0.0015 ) . Abnormal copy number of P35354 was marginally associated with objective response ( P = 0.066 ) , while abnormal copy numbers of two reference loci , 1q25 and the chromosome 10 centromere , were significantly associated with response . Pairwise combinations of copy number status of these loci and P04626 amplicon size provided stronger associations and identified a group of patients without responders . These results suggest that patient selection for trastuzumab may be improved by considering P04626 amplicon size and genomic status of the 1q25 , P35354 , and centromere 10 loci . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) A structural and in vitro characterization of asoprisnil : a selective progesterone receptor modulator . Selective progesterone receptor modulators ( SPRMs ) have been suggested as therapeutic agents for treatment of gynecological disorders . One such SPRM , asoprisnil , was recently in clinical trials for treatment of uterine fibroids and endometriosis . We present the crystal structures of progesterone receptor ( PR ) ligand binding domain complexed with asoprisnil and the corepressors nuclear receptor corepressor ( NCoR ) and Q9Y618 . This is the first report of steroid nuclear receptor crystal structures with ligand and corepressors . These structures show PR in a different conformation than PR complexed with progesterone ( P4 ) . We profiled asoprisnil in PR-dependent assays to understand further the PR-mediated mechanism of action . We confirmed previous findings that asoprisnil demonstrated antagonism , but not agonism , in a PR-B transfection assay and the T47D breast cancer cell alkaline phosphatase activity assay . Asoprisnil , but not DB00834 , weakly recruited the coactivators Q15788 and Q9Y6Q9 . However , asoprisnil strongly recruited the corepressor NCoR in a manner similar to DB00834 . Unlike DB00834 , NCoR binding to asoprisnil-bound PR could be displaced with equal affinity by NCoR or Q15596 peptides . We further showed that it weakly activated T47D cell gene expression of Sgk-1 and O60437 and antagonized P4-induced expression of both genes . In rat leiomyoma ELT3 cells , asoprisnil demonstrated partial P4-like inhibition of cyclooxygenase ( P36551 ) enzymatic activity and P35354 gene expression . In the rat uterotrophic assay , asoprisnil demonstrated no P4-like ability to oppose estrogen . Our data suggest that asoprisnil differentially recruits coactivators and corepressors compared to DB00834 or P4 , and this specific cofactor interaction profile is apparently insufficient to oppose estrogenic activity in rat uterus .

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interacts_with DB00188?

multiple_choice

Point mutation of the proteasome beta5 subunit gene is an important mechanism of bortezomib resistance in bortezomib-selected variants of Jurkat T cell lymphoblastic lymphoma/leukemia line . To study the mechanism of acquired resistance to bortezomib , a new antitumor drug that is the first therapeutic proteasome inhibitor , we established a series of bortezomib-resistant T lymphoblastic lymphoma/leukemia cell lines , designated the JurkatBs , from the parental Jurkat line via repeated drug selection . There were no significant differences in the growth curves or colony formation between the JurkatB cells and parental Jurkat cells . The effects of bortezomib on cytotoxicity , cell cycle arrest , and induction of apoptosis were decreased in JurkatB cells compared with parental Jurkat cells . A mutation in the proteasome beta5 subunit ( P28074 ) gene ( G322A ) , which encodes an amino acid change from Ala to DB00156 at polypeptide position 108 , was detected by sequencing full-length cDNA clones and direct polymerase chain reaction products of the P28074 gene . DB00188 caused less inhibition of chymotrypsin-like activity in resistant cells . When the G322A mutant P28074 was retrovirally introduced into parental Jurkat cells , it conferred bortezomib resistance to these cells , resulting in decreased cytotoxicity , apoptosis , and inhibition of chymotrypsin-like activity . The predicted structure of A108T-mutated P28074 shows a conformational change that suggests decreased affinity to bortezomib . In short , the G322A mutation of the P28074 gene is a novel mechanism for bortezomib resistance . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Potent and selective activation of the pancreatic beta-cell type K( DB00171 ) channel by two novel diazoxide analogues . AIMS/HYPOTHESIS : We investigated the pharmacological properties of two novel DB00171 sensitive potassium ( K( DB00171 ) ) channel openers , 6-Chloro-3-isopropylamino-4 H-thieno [ 3,2- e ] -1,2,4-thiadiazine 1,1-dioxide ( NNC 55-0118 ) and 6-chloro-3-(1-methylcyclopropyl)amino-4 H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide ( NN414 ) , on the cloned cardiac ( Kir6.2/SUR2A ) , smooth muscle ( Kir6.2/SUR2B ) and pancreatic beta cell ( Kir6.2/ Q09428 ) types of K( DB00171 ) channel . METHODS : We studied the effects of these compounds on whole-cell currents through cloned K( DB00171 ) channels expressed in Xenopus oocytes or mammalian cells ( HEK293 ) . We also used inside-out macropatches excised from Xenopus oocytes . RESULTS : In P29320 293 cells , NNC 55-0118 and NN414 activated Kir6.2/ Q09428 currents with EC(50) values of 0.33 micromol/l and 0.45 micromol/l , respectively , compared with that of 31 micro mol/l for diazoxide . Neither compound activated Kir6.2/SUR2A or Kir6.2/SUR2B channels expressed in oocytes , nor did they activate Kir6.2 expressed in the absence of Q09428 . Current activation was dependent on the presence of intracellular MgATP , but was not supported by MgADP . CONCLUSION/INTERPRETATION : Both NNC 55-0118 and NN414 selectively stimulate the pancreatic beta-cell type of K( DB00171 ) channel with a higher potency than diazoxide , by interaction with the Q09428 subunit . The high selectivity and efficacy of the compounds could prove useful for treatment of disease states where inhibition of insulin secretion is beneficial . Experimental Staphylococcus aureus infection of the mammary gland induces region-specific changes in innate immune gene expression . Staphylococcus aureus is a prolific mastitis-causing bacterium that resides naturally in the environment of the dairy cow . The aim of this study was to profile immune gene expression in tissue from the alveolar , ductal , gland cistern and teat canal regions of the bovine mammary gland following intramammary infection with S. aureus . Quantitative real-time PCR ( qPCR ) was used to profile expression of innate immune genes including pattern recognition receptors ( PRRs ) , cytokines , antimicrobial peptides ( AMPs ) and acute phase proteins ( APPs ) . Consistent expression of Toll-like receptors ( TLRs ) 1-10 and NOD-like receptors ( NODs ) 1-2 was detected in all four tissue regions . Pro-inflammatory cytokines ( P05231 , Q16552 and P10145 ) and anti-inflammatory cytokine ( P22301 ) were induced in all 4 tissues . P05067 ( SAA3 and HP ) and AMP ( DEFB4 and Q8NG35 ) genes showed the greatest induction throughout the mammary gland in response to S. aureus , with particularly high expression in alveolar tissue ( SAA3 and HP > 133- and > 80-fold respectively , P < 0.05 ; DEFB4 and Q8NG35 > 9- and > 27-fold respectively , P < 0.05 ) . Collectively , our data show both sentinel and effector immune functions throughout the mammary gland in response to S. aureus challenge . Variability in the 5-HT(2A) receptor gene is associated with seasonal pattern in major depression . The 102-T/C polymorphism of the 5-HT(2A) receptor gene was analysed in 159 patients with major depression and 164 unrelated and healthy controls using a case-control design . Allele and genotype frequencies did not differ between cases and controls . No differences according to sex , age of onset , melancholia , suicidal behaviour or family history of psychiatric illness were found . However , genotype distributions significantly differed between patients with seasonal pattern in their episodes ( P43034 ) and patients with no seasonal pattern ( N- P43034 ) ( chi(2) = 10.63 ; P = 0.004 ) . A seasonal pattern was 7.57 times more frequent in 102C-allele carriers than in 102T homozygous ( 95.1 % of patients P43034 carried 102C-allele vs 72 % of patients N- P43034 ( chi(2) = 9.45 , df=1 , P = 0.002 ; OR = 7.57 ( 95 % CI : 1.65 -- 48.08 ) ) . These results suggest that variation in the 5- Q13049 receptor gene may play a role in the development of major depression with seasonal pattern and support the existence of a genetic and etiological heterogeneity underlying the diagnosis of major depression . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . DB00188 -resistant myeloma cell lines : a role for mutated P28074 in preventing the accumulation of unfolded proteins and fatal ER stress . DB00188 is an effective agent for treating multiple myeloma ( MM ) . To investigate the underlying mechanisms associated with acquired resistance to this agent , we established two bortezomib-resistant MM cell lines , KMS-11/BTZ and OPM-2/BTZ , the 50 % inhibitory concentration values of which were respectively 24.7- and 16.6-fold higher than their parental cell lines . No activation of caspase and BH3-only proteins such as Noxa was noted in bortezomib-resistant cells after exposure to the drug . The accumulation of polyubiquitinated proteins was reduced in bortezomib-resistant cells compared with the parental cells , associated with avoidance of catastrophic ER stress as assessed by downregulation of P35638 expression . These resistant MM cells have a unique point mutation , G322A , in the gene encoding the proteasome beta5 subunit ( P28074 ) , likely resulting in conformational changes to the bortezomib-binding pocket of this subunit . KMS-11 parental cells transfected to express mutated P28074 also showed reduced bortezomib-induced apoptosis compared with those expressing wild-type P28074 or the parental cells . Expression of mutated P28074 was associated with the prevention of the accumulation of unfolded proteins . Thus , a fraction of MM cells may acquire bortezomib resistance by suppressing apoptotic signals through the inhibition of unfolded protein accumulation and subsequent excessive ER stress by a mutation of the P28074 gene . Immunoaffinity purification of the functional 20S proteasome from human cells via transient overexpression of specific proteasome subunits . The proteasome is a multi-subunit proteolytic complex that plays a central role in protein degradation in all eukaryotic cells . It regulates many vital cellular processes therefore its dysfunction can lead to various pathologies including cancer and neurodegeneration . Isolation of enzymatically active proteasomes is a key step to the successful study of the proteasome regulation and functions . Here we describe a simple and efficient protocol for immunoaffinity purification of the functional 20S proteasomes from human P29320 293T cells after transient overexpression of specific proteasome subunits tagged with 3xFLAG . To construct 3xFLAG-fusion proteins , DNA sequences encoding the 20S proteasome subunits P28074 , P28066 , and P25788 were cloned into mammalian expression vector pIRES-hrGFP-1a . The corresponding recombinant proteins P28074 -3xFLAG , P28066 -3xFLAG , or P25788 -3xFLAG were transiently overexpressed in human P29320 293T cells and were shown to be partially incorporated into the intact proteasome complexes . 20S proteasomes were immunoprecipitated from P29320 293T cell extracts under mild conditions using antibodies against FLAG peptide . Isolation of highly purified 20S proteasomes were confirmed by SDS-PAGE and Western blotting using antibodies against different proteasome subunits . Affinity purified 20S proteasomes were shown to possess chymotrypsin- and trypsin-like peptidase activities confirming their functionality . This simple single-step affinity method of the 20S proteasome purification can be instrumental to subsequent functional studies of proteasomes in human cells . Proteasomal serine hydrolases are up-regulated by and required for influenza virus infection . Interactions between viruses and their host cells are important determinants of virus replication and of immune responses to the virus . However , these interactions and resulting consequences of these interactions remain poorly defined . Numerous recent quantitative proteomic approaches have measured host proteins affected by virus infection . Here , we used activity-based protein profiling ( P05067 ) to measure functional alterations in host serine hydrolases after influenza A virus infection of Madin-Darby canine kidney and human A549 lung cells . We identified 62 serine proteases . We then combined the P05067 approach with stable isotope labeling to directly measure how serine hydrolase activities were affected by virus infection . Differentially regulated SHs mapped into a few key cellular pathway systems , most notably the proteasomal system . The specific serine protease inhibitors DB06692 and Pefablock and specific proteasomal inhibitors DB00188 and MG132 significantly inhibited influenza virus growth . Some inhibitors also down-regulated activities of several proteasomal proteins , including P25786 , P25787 , and PMSB3 . Genetic knockdown of PMSA2 also attenuated influenza virus replication . These findings further our understanding of enzymatic cellular processes affected by influenza virus and may be beneficial in the search for additional antiviral therapeutic targets . Alzheimer 's disease and modern lifestyle : what is the role of stress ? This Editorial highlights a study by Baglietto-Vargas et al. 2015 published in this issue of J. Neurochem . Stress is one of the environmental factors that can contribute to Alzheimer 's disease pathogenesis . However , the role of modern-life stress has not been investigated yet . The authors reveal that modern-life stress reduces the number of dendritic spines in the hippocampus of Alzheimer 's disease transgenic mice . The mechanism underlying such effect involves an increase in corticotropin-releasing hormone ( P06850 ) release that stimulates the amyloid precursor protein ( P05067 ) processing and fosters the generation of Amyloid-β , which negatively affects dendritic spines . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . No evidence of mutations of the P28074 ( beta-5 subunit of proteasome ) in a case of myeloma with clinical resistance to DB00188 . Evaluation of hypoxia inducible factor expression in inflammatory and neurodegenerative brain models . The neuroinflammatory process is thought to contribute to the progression of neurological disorders and brain pathologies . The release of pro-inflammatory cytokines and chemokines by activated glial cells , astrocytes and microglia plays an important role in this process . However , the role of hypoxia-inducible factor-1α ( HIF-1α ) , the key transcription factor regulating the expression of hypoxia-inducible genes , during glial activation is less known . Thus , we examined the significance of HIF-1α in three experimental models : first in an acute model of inflammation induced by pro-inflammatory cytokines P01375 -α , IL-1β and IFN-γ ; secondly , in a chronic model of inflammation using an APPswe/PS1dE9 ( P05067 / P49768 ) transgenic mouse model of Alzheimer 's disease and thirdly via the inhibition of the PI3K/AKT pathway in a model of neuronal apoptosis . During acute glial inflammation induced by in vitro administration of P01375 -α , IL-1β and IFN-γ , mRNA expression levels of HIF-1α were significantly upregulated ; however , this effect was blocked by SP600126 , a pharmacological inhibitor of mitogen-activated protein kinases ( MAPKs ) . These data suggest that MAPKs could be involved in HIF-1α regulation . In addition , we observed that HIF-1α is not involved in the neuronal apoptotic process mediated by P19957 -kinase inhibition , which is regulated by c-Jun . Finally , we did not detect significant differences in the expression of HIF-1α mRNA in P05067 / P49768 mice during the course of the study ( 3-12 months of age ) . Thus , we demonstrated that HIF-1α has a prominent role in acute but not in chronic inflammatory processes , such as the one which occurs in the P05067 / P49768 experimental model of AD . Moreover , HIF-1α is not involved in neuronal apoptosis after PI3K/AKT inhibition . Spinal cord injury induces early and persistent lesional Q99571 receptor expression . Following spinal cord injury ( SCI ) , neuropathic , chronic pain is a major cause of disability . Recently , glial Q99571 receptor ( P2X4R ) has been identified as a major contributor to the development of neuropathic pain after peripheral nerve injury . Here we report analysis of P2X4R expression following rat SCI . Significant lesional accumulation of P2X4R+ cells was detected as early as 24 h after SCI , reaching maximum cell numbers on Day 7 . Thereafter cell numbers declined but persisted at significantly elevated , sub-maximal levels ( > 70 % ) until 1 month post injury . Double-immunolabeling identified the majority of lesional P2X4R+ cells as activated microglia/macrophages and surviving neurons/neurites . Increase of P2X4R+ , beta- P05067 + hypertrophic neurites correlated with proximity to the lesion . Further , P2X4R+ cells coexpressed the intracellular regulators of signalling cascades , P23219 ( > 20 % ) , P35354 ( > 5 % ) , RhoA ( > 60 % ) and RhoB ( > 10 % ) . G(alpha)12/13 inhibition enhances the anticancer effect of bortezomib through P28074 downregulation . DB00188 is a proteasome inhibitor approved for anticancer therapy . However , variable sensitivity of tumor cells exists in this therapy probably due to differences in the expression of proteasome subunits . G(alpha)(12/13) serves modulators or signal transducers in diverse pathways . This study investigated whether cancer cells display differential sensitivity to bortezomib with reference to G(alpha)(12/13) expression , and if so , whether G(alpha)(12/13) affects the expression of proteasome subunits and their activities . DB00188 treatment exhibited greater sensitivities in Huh7 and SNU886 cells ( epithelial type ) than SK-Hep1 and SNU449 cells ( mesenchymal type ) that exhibited higher levels of G(alpha)(12/13) . Overexpression of an active mutant of G(alpha)(12) ( Galpha(12)QL ) or G(alpha)(13) ( G(alpha)(13)QL ) diminished the ability of bortezomib to induce cytotoxicity in Huh7 cells . Moreover , transfection with the minigene that disturbs G protein-coupled receptor-G protein coupling ( CT12 or Q9NXZ2 ) increased it in SK-Hep1 cells . Consistently , MiaPaCa2 cells transfected with CT12 or Q9NXZ2 exhibited a greater sensitivity to bortezomib . Evidence of G(alpha)(12/13) 's antagonism on the anticancer effect of bortezomib was verified in the reversal by G(alpha)(12)QL or G(alpha)(13)QL of the minigenes ' enhancement of cytotoxity . Real-time polymerase chain reaction assay enabled us to identify P28074 , multicatalytic endopeptidase complex-like-1 , and proteasome activator subunit-1 repression by CT12 or Q9NXZ2 . Furthermore , G(alpha)(12/13) inhibition enhanced the ability of bortezomib to repress P28074 , as shown by immunoblotting and proteasome activity assay . Moreover , this inhibitory effect on P28074 was attenuated by G(alpha)G(alpha)(12)QL or G(alpha)(13)QL . In conclusion , the inhibition of G(alpha)(12/13) activities may enhance the anticancer effect of bortezomib through P28074 repression , providing insight into the G(alpha)(12/13) pathway for the regulation of proteasomal activity . Enhanced goblet cell hyperplasia in HDC knockout mice with allergic airway inflammation . BACKGROUND : DB11320 is known to have immunoregulatory roles in allergic reactions through histamine receptor 1 ( P35367 ) , P25021 , Q9Y5N1 and Q9H3N8 . However , its role in goblet cell hyperplasia in the airways of asthma patients is yet to be clarified . OBJECTIVE : This study was designed to examine the role of histamine in goblet cell hyperplasia using histamine-deficient mice ( Hdc-/- mice ) with allergic airway inflammation . METHODS : Wild-type and Hdc-/- C57BL/6 mice were sensitized with ovalbumin ( OVA ) . After a 2-week exposure to OVA , goblet cell hyperplasia was evaluated . Cell differentials and cytokines in BALF were analyzed . The mRNA levels of P98088 and Gob-5 gene were determined quantitatively . RESULTS : The number of eosinophils in BALF increased in both the sensitized wild-type mice and Hdc-/- mice with OVA inhalation . In addition , the numbers of alveolar macrophages and lymphocytes in BALF increased significantly in the sensitized Hdc-/- mice with OVA inhalation compared to the wild-type mice under the same conditions . The concentrations of P05112 ( P05112 ) , P05113 , P35225 , Interferon-gamma ( P01579 ) , tumor necrosis factor-alpha ( P01375 ) and P60568 in the BALF all increased significantly in both groups compared to those exposed to saline . In particular , the concentration of P01375 in the Hdc-/- mice exposed to OVA was significantly higher than that in the wild-type mice under the same conditions . The mRNA levels of Gob-5 and P98088 , and the ratio of the goblet cells in the airway epithelium significantly increased in Hdc-/- mice exposed to OVA compared to wild-type mice . CONCLUSIONS : These results suggested that histamine may play a regulatory role in goblet cell hyperplasia in allergic airway inflammation . Knockdown of human deubiquitinase O00487 induces cell cycle arrest and senescence . The O00487 ( O00487 , also known as Rpn11/MPR1/S13/CepP1 ) protein within the 19S complex ( 19S cap ; PA700 ) is responsible for substrate deubiquitination during proteasomal degradation . The role of O00487 in cell proliferation and senescence was explored using siRNA knockdown in carcinoma cell lines . Our results reveal that down-regulation of O00487 by siRNA transfection had a considerable impact on cell viability causing cell arrest in the G0- P55008 phase , ultimately leading to senescence . The molecular events associated with decreased cell proliferation , cell cycle arrest and senescence include down-regulation of cyclin B1- P06493 - P30307 , down-regulation of cyclin D1 and up-regulation of P38936 (/Cip) and p27(/Kip1) . Most notably , phosphorylation of the retinoblastoma protein was markedly reduced in O00487 knockdown cells . A comparative study with P28074 , a subunit of the 20S proteasome , revealed that P28074 and O00487 have different effects on cell cycle , senescence and associated molecular events . These data support the view that the 19S and 20S subunits of the proteasome have distinct biological functions and imply that targeting 19S and 20S would have distinct molecular consequences on tumor cells . Characterization of bortezomib-adapted I-45 mesothelioma cells . BACKGROUND : DB00188 , a proteasome-specific inhibitor , has emerged as a promising cancer therapeutic agent . However , development of resistance to bortezomib may pose a challenge to effective anticancer therapy . Therefore , characterization of cellular mechanisms involved in bortezomib resistance and development of effective strategies to overcome this resistance represent important steps in the advancement of bortezomib-mediated cancer therapy . RESULTS : The present study reports the development of I-45-BTZ-R , a bortezomib-resistant cell line , from the bortezomib-sensitive mesothelioma cell line I-45 . I-45-BTZ-R cells showed no cross-resistance to the chemotherapeutic drugs cisplatin , 5-fluorouracil , and doxorubicin . Moreover , the bortezomib-adapted I-45-BTZ-R cells had decreased growth kinemics and did not over express proteasome subunit beta5 ( P28074 ) as compared to parental I-45 cells . I-45-BTZ-R cells and parental I-45 cells showed similar inhibition of proteasome activity , but I-45-BTZ-R cells exhibited much less accumulation of ubiquitinated proteins following exposure to 40 nm bortezomib . Further studies revealed that relatively low doses of bortezomib did not induce an unfolded protein response ( UPR ) in the bortezomib-adapted cells , while higher doses induced UPR with concomitant cell death , as evidenced by higher expression of the mitochondrial chaperone protein Bip and the endoplasmic reticulum ( ER ) stress-related pro-apoptotic protein P35638 . In addition , bortezomib exposure did not induce the accumulation of the pro-apoptotic proteins p53 , Mcl-1S , and noxa in the bortezomib-adapted cells . CONCLUSION : These results suggest that UPR evasion , together with reduced pro-apoptotic gene induction , accounts for bortezomib resistance in the bortezomib-adapted mesothelioma cell line I-45-BTZ-R . Cytotoxic effects of bortezomib in myelodysplastic syndrome/acute myeloid leukemia depend on autophagy-mediated lysosomal degradation of Q9Y4K3 and repression of P25786 . DB00188 ( Velcade ) is used widely for the treatment of various human cancers ; however , its mechanisms of action are not fully understood , particularly in myeloid malignancies . DB00188 is a selective and reversible inhibitor of the proteasome . Paradoxically , we find that bortezomib induces proteasome-independent degradation of the Q9Y4K3 protein , but not mRNA , in myelodysplastic syndrome ( P43034 ) and acute myeloid leukemia ( AML ) cell lines and primary cells . The reduction in Q9Y4K3 protein coincides with bortezomib-induced autophagy , and subsequently with apoptosis in P43034 /AML cells . RNAi-mediated knockdown of Q9Y4K3 sensitized bortezomib-sensitive and -resistant cell lines , underscoring the importance of Q9Y4K3 in bortezomib-induced cytotoxicity . DB00188 -resistant cells expressing an shRNA targeting Q9Y4K3 were resensitized to the cytotoxic effects of bortezomib due to down-regulation of the proteasomal subunit α-1 ( P25786 ) . To determine the molecular consequences of loss of Q9Y4K3 in P43034 /AML cells , in the present study , we applied gene-expression profiling and identified an apoptosis gene signature . Knockdown of Q9Y4K3 in P43034 /AML cell lines or patient samples resulted in rapid apoptosis and impaired malignant hematopoietic stem/progenitor function . In summary , we describe herein novel mechanisms by which Q9Y4K3 is regulated through bortezomib/autophagy-mediated degradation and by which it alters P43034 /AML sensitivity to bortezomib by controlling P25786 expression . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Rational approaches to design of therapeutics targeting molecular markers . This paper introduces novel therapeutic strategies focusing on a molecular marker relevant to a particular hematologic malignancy . Four different approaches targeting specific molecules in unique pathways will be presented . The common theme will be rational target selection in a strategy that has reached the early phase of human clinical trial in one malignancy , but with a much broader potential applicability to the technology . In Section I Dr. Richard Klasa presents preclinical data on the use of antisense oligonucleotides directed at the bcl-2 gene message to specifically downregulate Bcl-2 protein expression in non-Hodgkin 's lymphomas and render the cells more susceptible to the induction of apoptosis . In Section II Dr. Alan List reviews the targeting of vascular endothelial growth factor ( P15692 ) and its receptor in anti-angiogenesis strategies for acute myeloid leukemia ( AML ) and myelodysplastic syndromes ( P43034 ) . In Section III Dr. Bruce Cheson describes recent progress in inhibiting cell cycle progression by selectively disrupting cyclin D1 with structurally unique compounds such as flavopiridol in mantle cell lymphoma as well as describing a new class of agents that affect proteasome degradation pathways . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Biological and clinical relevance of matrix metalloproteinases 2 and 9 in acute myeloid leukaemias and myelodysplastic syndromes . We analysed by immunocytochemistry metalloproteinase ( MMP ) -2 and P14780 expression in bone marrow cells from 54 acute myeloid leukaemia ( AML ) patients , 153 myelodysplastic syndrome ( P43034 ) patients , and 52 non-haemopathic subjects , in order to evaluate whether MMP expression abnormalities were associated with relevant laboratory or clinical findings . In normal samples P08253 was detected in rare myeloid cells , P14780 in most maturing myeloid cells . In P43034 P08253 myeloid levels were higher than in controls ( P < 0.0001 ) ; P08253 and P14780 were often co-expressed . Also many erythroblasts expressed P08253 . There was a positive correlation between P08253 erythroblast expression and erythroid dysplasia ( P = 0.002 ) and an inverse correlation between P08253 or P14780 myeloid expression and blast cell percentage ( P = 0.05 and P = 0.04 respectively ) . High MMP levels in myeloid cells were associated with longer overall survival ( P = 0.03 ) and evolution-free survival ( P = 0.04 ) . In AML P08253 levels were lower than in P43034 ( P < 0.0001 ) and P14780 levels lower than in P43034 and controls ( P < 0.0001 ) . MMP levels did not predict response to therapy . The release of active MMPs was detected by colorimetric analysis in cell cultures from representative P43034 and AML cases . In conclusion , we have demonstrated an abnormal MMP expression in AML as well as in P43034 . The production and release of these enzymes may influence haematopoietic cell behaviour . In P43034 , the detection of MMP deregulated expression may be important also from the clinical point of view : it may provide a useful tool for diagnosis , prognosis and a possible target for experimental treatments . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . P15692 -induced angiogenesis ameliorates the memory impairment in P05067 transgenic mouse model of Alzheimer 's disease . Vascular endothelial growth factor ( P15692 ) was investigated in the present study to see whether it could provide a therapeutic opportunity for the treatment of Alzheimer 's disease ( AD ) . PDGF-hAPP(V717I) transgenic mice were treated with P15692 or PBS by intraperitoneal injection for three consecutive days . The results showed that P15692 ameliorated the memory impairment of mice , accompanied by P28906 (+) cells increasing in peripheral blood , P04275 (+) vessels increasing in hippocampus , and P28906 (+)/ P35968 (+) , P04275 (+)/ P35968 (+) and BrdU(+)/ P04275 (+) cells expressing in hippocampus . Furthermore , the level of choline acetyltransferase ( P28329 ) was considerably enhanced and Aβ deposition was decreased in the brains of mice upon P15692 treatment . These observations suggest that P15692 should be pursued as a novel therapeutic agent for treatment of AD . Structural analysis and chromosomal localization of the mouse Psmb5 gene coding for the constitutively expressed beta-type proteasome subunit . The proteasome is a multi-subunit protease responsible for the production of peptides presented by major histocompatibility complex class I molecules . Accumulated evidence indicates that , upon stimulation with interferon-gamma ( P01579 ) , three beta-type subunits , designated P28065 , P28062 , and P40306 , are incorporated into the 20S proteasome by displacing the housekeeping beta-type subunits designated P28072 , P28074 , and Q99436 , respectively . These changes in the subunit composition appear to facilitate class I-mediated antigen presentation , presumably by altering the cleavage specificities of the proteasome . In the present study , we determined the organization of the mouse gene Psmb5 , coding for the P28074 subunit . Psmb5 is made up of three exons , spanning approximately 5 kilobases . Its exon-intron organization differs radically from those of the other P01579 -regulated , beta-type subunit genes including Lmp7 with which Psmb5 is believed to share an immediate common ancestor . The structure of the mouse Psmb5 gene is identical to that of its recently characterized human counterpart . Thus , the unique organization of the gene coding for the P28074 subunit appears to have been established before mammalian radiation . As well as the Psmb5 gene , the mouse genome contains a processed pseudogene designated Psmb5-ps . Interspecific backcross mapping showed that Psmb5 maps close to the Gtrgal2 locus on chromosome 14 and that Psmb5-ps is located in the vicinity of the Psme3 locus on chromosome 11 . These results were confirmed by fluorescent in situ hybridization analysis that localized Psmb5 to band P06681 to proximal D1 of chromosome 14 and Psmb5-ps to band D of chromosome 11 . DB00104 -targeted liposomes loaded with CPT-11 enhanced cytotoxicity for the treatment of medullary thyroid carcinoma . Medullary thyroid carcinoma ( P04629 ) is a rare endocrine tumor that frequently metastasizes , but treatment with irinotecan ( CPT-11 ) is limited because of side effects . P04629 is known to overexpress the somatostatin receptor subtype 2 ( P30874 ) . DB00104 ( Oct ) is a somatostatin analogue that has a high binding affinity for SSTR and can be used as a tumor-targeting ligand . We prepared Oct-targeted liposomes loaded with CPT-11 using Oct-poly ( ethylene glycol ) ( PEG ) -lipid and evaluated Oct-mediated association and cytotoxicity of the liposomes with an P04629 cell line TT . The association of higher concentrations of modified Oct-targeted liposomes with TT cells was significantly higher than PEGylated liposomes and was significantly inhibited by empty Oct-targeted liposomes but not by free Oct . With exposure for 96 h , the cytotoxicity of Oct-targeted liposomal CPT-11 ( IC50 : 1.05 ± 0.47 μM ) was higher than free CPT-11 ( IC50 : 3.76 ± 0.61 μM ) or PEGylated liposomal CPT-11 ( IC50 : 3.05 ± 0.28 μM ) . In addition , empty Oct-targeted liposomes showed significantly higher cytotoxicity than empty nontargeted liposomes at a concentration where free Oct did not show cytotoxicity , suggesting that Oct as a ligand showed cytotoxicity . Moreover , Oct-targeted liposomal CPT-11 led to significantly higher antitumor activity and prolonged the survival time compared with nontargeted liposomal and free CPT-11 at a one-third dose and lower administration times with free CPT-11 . These findings indicated that Oct-targeted liposomes loaded with CPT-11 may offer considerable potential for P04629 chemotherapy because cytotoxicity of both CPT-11 and Oct was enhanced by effective cellular uptake via P30874 . P05067 is a primary androgen target gene that promotes prostate cancer growth . P10275 ( AR ) is a critical transcription factor that regulates various target genes and contributes to the pathophysiology of prostate cancer hormone dependently . Here , we identify amyloid precursor protein ( P05067 ) as a primary androgen target through chromatin immunoprecipitation ( ChIP ) combined with genome tiling array analysis ( ChIP-chip ) . ChIP-treated DNA were obtained from prostate cancer LNCaP cells with R1881 or vehicle treatment using AR or acetylated histone H3 antibodies . Ligand-dependent AR binding was further enriched by PCR subtraction . Using chromosome 21/22 arrays , we identified P05067 as one of the androgen-regulated genes with adjacent functional AR binding sites . P05067 expression is androgen-inducible in LNCaP cells and P05067 immunoreactivity was correlated with poor prognosis in patients with prostate cancer . Gain-of-function and loss-of-function studies revealed that P05067 promotes the tumor growth of prostate cancer . The present study reveals a novel P05067 -mediated pathway responsible for the androgen-dependent growth of prostate cancer . Our findings will indicate that P05067 could be a potential molecular target for the diagnosis and treatment of prostate cancer . Regulation of P28074 protein and β subunits of mammalian proteasome by constitutively activated signal transducer and activator of transcription 3 ( P40763 ) : potential role in bortezomib-mediated anticancer therapy . The ubiquitin-proteasome system facilitates the degradation of ubiquitin-tagged proteins and performs a regulatory role in cells . Elevated proteasome activity and subunit expression are found in several cancers . However , the inherent molecular mechanisms responsible for increased proteasome function in cancers remain unclear despite the well investigated and defined role of the mammalian proteasome . This study was initiated to elucidate the mechanisms involved in the regulation of β subunits of the mammalian proteasome . Suppression of P40763 tyrosine phosphorylation coordinately decreased the mRNA and protein levels of the β subunits of the 20 S core complex in DU145 cells . Notably , P28074 , a molecular target of bortezomib , was shown to be a target of P40763 . Knockdown of P40763 decreased P28074 protein . Inhibition of phospho- P40763 substantially reduced P28074 protein levels in cells expressing constitutively active- P40763 . Accumulation of activated P40763 resulted in the induction of P28074 promoter and protein levels . In addition , a direct correlation was observed between the endogenous levels of P28074 and constitutively active P40763 . P28074 and P40763 protein levels remained unaltered following the inhibition of proteasome activity . The P01133 -induced concerted increase of β subunits was blocked by inhibition of the P01133 receptor or P40763 but not by the PI3K/AKT or MEK/ P29323 pathways . Decreased proteasome activities were due to reduced protein levels of catalytic subunits of the proteasome in P40763 -inhibited cells . Combined treatments with bortezomib and inhibitor of P40763 abrogated proteasome activity and enhanced cellular apoptosis . Overall , we demonstrate that aberrant activation of P40763 regulates the expression of β subunits , in particular P28074 , and the catalytic activity of the proteasome . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . Cellular responses to murine P25942 in a mouse B cell line may be TRAF dependent or independent . Engagement of P25942 by its ligand induces IKK and mitogen-activated protein kinase ( MAPK ) phosphorylation and transcriptional activation , leading to activation and differentiation of B cells . These events are most likely transduced by adaptor molecules that are recruited to the P25942 cytoplasmic domain , called P01375 receptor-associated factors ( TRAF ) . We have engineered a chimeric P25942 molecule using the human extracellular sequence and the murine cytoplasmic domain to assess the contribution that specific TRAF binding domains provide to the cytoplasmic signaling functions of P25942 . The data presented here show that the shared binding site for TRAF2 and Q13114 accounts for receptor internalization , and the majority of signaling through P25942 , but is redundant with the Q9Y4K3 binding site for activation of p38 and NFkappaB signaling pathways . Disruption of the TRAF2/3 binding site results in a delayed and diminished kinase pathway induction , but complete preclusion of all signals requires the disruption of more than the two known TRAF binding sites . The specific TRAF dependency of P25942 -induced growth arrest , P01375 production , and phosphorylation of signaling molecules are shown , while p38 MAPK activation and cell surface antigen modulation suggest TRAF independent P25942 signaling in B cells . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Epigenetic control of aquaporin 1 expression by the amyloid precursor protein . Cellular processing of the amyloid precursor protein ( P05067 ) has been extensively studied , but its precise function remains elusive . The intracellular domain of P05067 has been proposed to regulate expression of several genes by mechanisms that are largely unknown . We report that P05067 regulates expression of the aquaporin 1 ( P29972 ) gene in mouse embryonic fibroblasts and in transgenic mice . P29972 mRNA and protein were down-regulated in fibroblasts lacking P05067 or presenilin 2 in which P29972 expression was restored by stable expression of full-length P05067 or presenilin 2 but not by P05067 deleted from its carboxy-terminal domain . The transcriptional activity of the P29972 gene promoter and the stability of P29972 mRNA were identical in fibroblasts expressing or not expressing P05067 . Control of P29972 expression by P05067 was sensitive to trichostatin A , an histone deacetylase inhibitor , and histone deacetylase activity coimmunoprecipitated with P05067 . Altogether , these data show that a presenilin-2-dependent gamma-secretase activity releases the intracellular domain of P05067 involved in the epigenetic control of P29972 expression . Since P29972 is found in astrocytes surrounding senile plaques , this epigenetic control of P29972 expression could have important implications in Alzheimer disease . Negative regulation of O00206 signaling by P22301 -dependent microRNA-146b . Toll-like receptors ( TLRs ) play key roles in detecting pathogens and initiating inflammatory responses that , subsequently , prime specific adaptive responses . Several mechanisms control TLR activity to avoid excessive inflammation and consequent immunopathology , including the anti-inflammatory cytokine P22301 . Recently , several TLR-responsive microRNAs ( miRs ) have also been proposed as potential regulators of this signaling pathway , but their functional role during the inflammatory response still is incompletely understood . In this study , we report that , after LPS engagement , monocytes up-regulate miR-146b via an P22301 -mediated P40763 -dependent loop . We show evidence that miR-146b modulates the O00206 signaling pathway by direct targeting of multiple elements , including the LPS receptor O00206 and the key adaptor/signaling proteins myeloid differentiation primary response ( MyD88 ) , interleukin-1 receptor-associated kinase 1 ( P51617 ) , and Q9Y4K3 ( Q9Y4K3 ) . Furthermore , we demonstrate that the enforced expression of miR-146b in human monocytes led to a significant reduction in the LPS-dependent production of several proinflammatory cytokines and chemokines , including P05231 , P01375 -α , P10145 , P10147 , P13500 , P80098 , and P02778 . Our results thus identify miR-146b as an P22301 -responsive miR with an anti-inflammatory activity based on multiple targeting of components of the O00206 pathway in monocytes and candidate miR-146b as a molecular effector of the P22301 anti-inflammatory activity . Euscaphic acid isolated from roots of Rosa rugosa inhibits LPS-induced inflammatory responses via O00206 -mediated NF-κB inactivation in RAW 264.7 macrophages . As an attempt to search for bioactive natural products exerting anti-inflammatory activity , we have evaluated the anti-inflammatory effects of euscaphic acid ( 19α-hydroxyursane-type triterpenoids , EA ) isolated from roots of Rosa rugosa and its underlying molecular mechanisms in lipopolysaccharide ( LPS ) -induced RAW 264.7 macrophages . EA concentration-dependently reduced the production of nitric oxide ( NO ) , prostaglandin E2 ( DB00917 ) , tumor necrosis factor-α ( P01375 -α ) , and interleukin-1β ( IL-1β ) induced by LPS in RAW 264.7 macgophages . Consistent with these data , expression levels of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) protein and P35228 , P35354 , P01375 -α , and IL-1β mRNA were inhibited by EA in a concentration-dependent manner . In addition , EA attenuated LPS-induced DNA binding and transcriptional activity of nuclear factor-kappa B ( NF-κB ) , which was accompanied by a parallel reduction of degradation and phosphorylation of inhibitory kappa Bα ( IκBα ) and consequently by decreased nuclear translocation of p65 subunit of NF-κB . Pretreatment with EA significantly inhibited the LPS-induced phosphorylation of IκB kinase β ( IKKβ ) , p38 , and JNK , whereas the phosphorylation of P27361 /2 was unaffected . Furthermore , EA interfered with the LPS-induced clustering of Q9Y4K3 ( Q9Y4K3 ) with interleukin receptor associated kinase 1 ( P51617 ) and transforming growth factor-β-activated kinase 1 ( TAK1 ) . Taken together , these results suggest that EA inhibits LPS-induced inflammatory responses by interference with the clustering of Q9Y4K3 with P51617 and TAK1 , resulting in blocking the activation of IKK and MAPKs signal transduction to downregulate NF-κB activations . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . Loss of P50281 causes cell senescence and nuclear defects which can be reversed by retinoic acid . P50281 ( P50281 ) is a collagenolytic enzyme located at the cell surface and implicated in extracellular matrix ( Q13201 ) remodeling . Mmp14(-/-) mice present dwarfism , bone abnormalities , and premature death . We demonstrate herein that the loss of P50281 also causes cardiac defects and severe metabolic changes , and alters the cytoskeleton and the nuclear lamina structure . Moreover , the absence of P50281 induces a senescent phenotype characterized by up-regulation of p16(INK4a) and P38936 (CIP1/WAF) ( 1 ) , increased activity of senescence-associated β-galactosidase , generation of a senescence-associated secretory phenotype , and somatotroph axis alterations . Consistent with the role of retinoic acid signaling in nuclear lamina stabilization , treatment of Mmp14(-/-) mice with all-trans retinoic acid reversed the nuclear lamina alterations , partially rescued the cell senescence phenotypes , ameliorated the pathological defects in bone , skin , and heart , and extended their life span . These results demonstrate that nuclear architecture and cell senescence can be modulated by a membrane protease , in a process involving the Q13201 as a key regulator of nuclear stiffness under cell stress conditions . The complete primary structure of mouse 20S proteasomes . The proteasome is a large multicatalytic proteinase that plays a role in the generation of peptides for presentation by major histocompatibility complex class I molecules . The 20S proteolytic core of mammalian proteasomes is assembled from a group of 17 protein subunits that generate a distinctive pattern of spots upon two-dimensional gel electrophoresis . The genes for most of these subunits have been cloned from humans and rats . We isolated cDNA clones for the mouse orthologues of ten of the subunits [ P25786 ( P06681 ) , P25787 ( P01024 ) , P25788 ( Q99618 ) , P25789 ( P02748 ) , P28066 ( ZETA ) , P60900 ( IOTA ) , O14818 ( P13671 -I ) , P49721 ( P10643 -I ) , P49720 ( Q99622 -II ) , and P28074 ( X ) ] to complete the cloning of all of the mouse subunits . Using antisera raised against these subunits or their orthologues , we verified the identity of these proteins by two-dimensional NEPHGE-PAGE . Analysis of a 26-kb region linked to the Mhc in zebrafish : genomic organization of the proteasome component beta/transporter associated with antigen processing-2 gene cluster and identification of five new proteasome beta subunit genes . Sequencing of zebrafish ( Danio rerio ) bacterial artificial chromosome and P1 artificial chromosome genomic clone fragments and of cDNA clones has led to the identification of five new loci coding for beta subunits of proteasomes ( PSMB ) . Together with the four genes identified previously , nine PSMB genes have now been defined in the zebrafish . Six of the nine genes reside in the zebrafish MHC ( Mhc ) class I region , four of them reside in a single cluster closely associated with TAP2 on a 26-kb long genomic fragment , and two reside at some distance from the fragment . In addition to homologues of the human genes P28074 through P28065 , two new genes , A5LHX3 and PSMB12 , have been found for which there are no known corresponding genes in humans . The new genes reside in the PSMB cluster in the Mhc . Homology and promoter region analysis suggest that the Mhc-associated genes might be inducible by P01579 . The zebrafish class I region contains representatives of three phylogenetically distinguishable groups of PSMB genes , X , Y , and Z . It is proposed that these genes were present in the ancestral PSMB region before Mhc class I genes became associated with it . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Computational insights into the selectivity mechanism of P05067 -IP over matrix metalloproteinases . In this work , selectivity mechanism of P05067 -IP inhibitor ( β-amyloid precursor protein-derived inhibitory peptide ) over matrix metalloproteinases ( MMPs including P08253 , P09237 , P14780 and P50281 ) was investigated by molecular modeling methods . Among MMPs , P08253 is the most favorable one for P05067 -IP interacting based on our calculations . The predicted binding affinities can give a good explanation of the activity difference of inhibitor P05067 -IP . In Comparison with P08253 / P05067 -IP complex , the side chain of Tyr214( P09237 ) makes the binding pocket so shallow that the whole side chain of Tyr3( P05067 -IP) can not be fully embraced , thus unfavorable for the N-terminal of P05067 -IP binding to P09237 . The poor selectivity of P05067 -IP toward P14780 is mainly related with the decrease of interaction between the P05067 -IP C-terminal and P14780 due to the bulky side chains of Pro193 and Gln199 , which is in agreement with experiment . The mutations at residues P193A and Q199G of P14780 alternate the binding pattern of the C-terminal of P05067 -IP by forming two new hydrogen bonds and hydrophobic interactions with P14780 . The mutants favor the binding affinity of P14780 largely . For P50281 / P05067 -IP , the large steric effect of Phe204( P50281 ) and the weak contributions of the polar residues Asn231( P50281 ) and Thr190( P50281 ) could explain why P50281 is non-selective for P05067 -IP interacting . Here , the molecular modeling methods were successfully employed to explore the selective inhibitor of MMPs , and our work gives valuable information for future rational design of selective peptide inhibitors toward individual MMP . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . DB02901 interacts with P00533 /MAPK signalling and modulates P00533 levels in androgen receptor-positive LNCaP prostate cancer cells . P10275 ( AR ) signalling plays a pivotal role in prostate cancer pathogenesis and progression . However , androgen-mediated AR signalling is yet to be fully understood . P00533 and Q96HU1 kinase signalling pathways play predominant roles in AR function . Therefore , we investigated the interaction of P00533 signalling and AR activity in AR-positive LNCaP cells . We found that 5alpha-dihydrotestosterone ( DB02901 ) and P01133 had a synergistic effect on AR activity as detected by a luciferase reporter system , although P01133 alone did not activate AR . Both P27361 /2 and p38 were involved in DB02901 and DB02901 / P01133 -induced AR activation as detected by specific MEK and p38 inhibitors . Furthermore , 24-h treatment of the cells with DB02901 resulted in ubiquitination and down-regulation of the P00533 . This effect could be inhibited by the anti-androgen flutamide , suggesting an androgen-dependent mechanism . On the other hand , DB02901 -treatment strongly increased AR levels in LNCaP cells . These data suggest a complex regulatory loop between activated AR and P00533 . In conclusion , activation of AR by both DB02901 and P01133 / DB02901 involves the Q96HU1 kinase pathway . Long-term activation of AR results in increase of AR levels , which through so far unknown regulatory mechanisms results in ubiquitination and degradation of the P00533 . Growth factor priming in therapy of acute myelogenous leukemia . Treatment of acute myelogenous leukemia ( AML ) incorporates the use of growth factors towards several objectives , including abbreviating the time of neutropenia , reducing susceptibility to infections , reducing length of hospitalization , and increasing susceptibility of the blasts to chemotherapy drugs , or priming . Priming is defined as the driving of leukemia cells into cell cycle in order to increase response to S-phase-specific drugs such as cytarabine ( DB00987 ) . Growth factors that have been used in priming include both filgrastim ( DB00099 ) and sagramostim ( GM- P04141 ) . Priming has often been applied in the treatment of older patients , who do not respond as well to standard treatment regimens , and whose disease was transformed from a myelodysplastic syndrome ( P43034 ) , or preleukemia . Recent , large randomized trials have demonstrated that although subgroups of patients may benefit , there does not appear to be any improvement in the complete remission rate sustained by inclusion of growth factor priming as part of the initial therapy of acute myelogenous leukemia . Hyperosmolar mannitol simulates expression of aquaporins 4 and 9 through a p38 mitogen-activated protein kinase-dependent pathway in rat astrocytes . The membrane pore proteins , aquaporins ( AQPs ) , facilitate the osmotically driven passage of water and , in some instances , small solutes . Under hyperosmotic conditions , the expression of some AQPs changes , and some studies have shown that the expression of P29972 and P55064 is regulated by MAPKs . However , the mechanisms regulating the expression of P55087 and AQP9 induced by hyperosmotic stress are poorly understood . In this study , we observed that hyperosmotic stress induced by mannitol increased the expression of P55087 and AQP9 in cultured rat astrocytes , and intraperitoneal infusion of mannitol increased P55087 and AQP9 in the rat brain cortex . In addition , a p38 MAPK inhibitor , but not P29323 and JNK inhibitors , suppressed their expression in cultured astrocytes . AQPs play important roles in maintaining brain homeostasis . The expression of P55087 and AQP9 in astrocytes changes after brain ischemia or traumatic injury , and some studies have shown that p38 MAPK in astrocytes is activated under similar conditions . Since mannitol is commonly used to reduce brain edema , understanding the regulation of AQPs and p38 MAPK in astrocytes under hyperosmotic conditions induced with mannitol may lead to a control of water movements and a new treatment for brain edema . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . The Q92734 protein , involved in oncogenic rearrangements , interacts with Q92844 and Q9Y6K9 , two proteins involved in the NF-kappaB pathway . P04629 -fused gene ( Q92734 ) was first identified as a partner of P04629 in generating the thyroid P04629 - DB00279 oncogene , and is also involved in oncogenic rearrangements with Q9UM73 in anaplastic lymphoma and NOR1 in mixoid chondrosarcoma . The Q92734 physiological role is still unknown , but the presence of a number of motifs involved in protein interactions suggests that it may function by associating with other proteins . We have recently demonstrated that Q92734 associates and regulates the activity of the tyrosine phosphatase Q15466 -1 . In this study by yeast two-hybrid screening we identified Q9Y6K9 and Q92844 , two proteins modulating the NF-kappaB pathway , as novel Q92734 -interacting proteins . These interactions were further characterized in vitro and in vivo . We provide evidence that Q92734 and Q9Y6K9 may be part of the same high molecular weight complex . Q92734 enhances the effect of P01375 , Q92844 , P01375 receptor-associated factor (TRAF)2 , and Q9Y4K3 in inducing NF-kappaB activity . We suggest that Q92734 is a novel member of the NF-kappaB pathway . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Association between Q06609 gene polymorphism ( -135G/C ) and susceptibility of myelodysplastic syndrome and acute leukemia : evidence based on a meta-analysis . Study results on the association between Q06609 gene -135G/C polymorphism and risk of myelodysplastic syndrome ( P43034 ) or acute leukemia are inconsistent . A meta-analysis was conducted to identify the association . A systematic search was performed in PubMed , Embase , CNKI , P01282 , Wanfang databases to collect all relevant studies until January 2013 . Meta-analysis was carried out using fixed/random model by Review Manager 5.1 and STATA10.0 . A total of 10 eligible studies with 2,656 patients and 3,725 controls were included in meta-analysis . Significant association was detected between -135G/C polymorphism and increased P43034 risk ( CC + GC vs. GG : OR = 1.46 , 95 % CI = 1.11-1.92 ; CC vs. GC + GG : OR = 2.45 , 95 % CI = 1.23-4.89 ) , while no association was observed for acute leukemia . Subgroup analysis by subtypes of acute leukemia and ethnicity showed no significant results either . Our meta-analysis indicated that the -135G/C polymorphism might be associated with increased susceptibility of P43034 . However , lack of evidence supported association of this polymorphism with acute leukemia . Additional well-designed studies with larger samples are required to verify our results . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) [ Effect of Jiawei Sini Decoction on Q9Y251 function of rats with restraint psychological stress ] . OBJECTIVE : To observe the effect of Jiawei Sini Decoction ( JWSND ) on Q9Y251 function of rats with restraint psychological stress and the pathway of thymus immunosuppressive function . METHODS : The rats were randomly divided into 4 groups , control group ( C ) , model group ( M ) , group treated by JWSND ( C1 ) and ginsenosides ( P06681 ) . Hypothalamus P06850 , blood plasma DB01285 and O00230 , the numberofthy moeyte GCR sites and the GCR nuclear translocation rate were detected by radioimmunoassay . RESULTS : Compared with C , in M , Hypothalamus P06850 , blood plasma DB01285 and O00230 increased significantly ( P < 0.01 ) , but all these of C1 and P06681 lowered significantly ( P < 0.05 , P < 0.01 ) . Compared with C , the GCR nuclear translocation rate was increased significantly ( P < 0. 01 ) . Compared with M , the GCR nuclear translocation rate lowered significantly ( P < 0.01 ) . Moreover , no significant difference was found in all indexes between C1 and C . CONCLUSION : JWSND participates effectively in the adjustment of Q9Y251 of the rats with restraint psychological stress , and relieves significantly the inhibitory effect of glucocorticoid on the thymus , the pathway closely relates to the translocation of GCR . P05067 -dependent alteration of GSK3β activity impairs neurogenesis in the Ts65Dn mouse model of Down syndrome . Intellectual disability in Down syndrome ( DS ) appears to be related to severe neurogenesis impairment during brain development . The molecular mechanisms underlying this defect are still largely unknown . Accumulating evidence has highlighted the importance of GSK3β signaling for neuronal precursor proliferation/differentiation . In neural precursor cells ( NPCs ) from Ts65Dn mice and human fetuses with DS , we found reduced GSK3β phosphorylation and , hence , increased GSK3β activity . In cultures of trisomic subventricular-zone-derived adult NPCs ( aNPCs ) we found that deregulation of GSK3β activity was due to higher levels of the AICD fragment of the trisomic gene P05067 that directly bound to GSK3β . We restored GSK3β phosphorylation in trisomic aNPCs using either lithium , a well-known GSK3β inhibitor , or using a 5-HT receptor agonist or fluoxetine , which activated the serotonin receptor P08908 . Importantly , this effect was accompanied by restoration of proliferation , cell fate specification and neuronal maturation . In agreement with results obtained in vitro , we found that early treatment with fluoxetine , which was previously shown to rescue neurogenesis and behavior in Ts65Dn mice , restored GSK3β phosphorylation . These results provide a link between GSK3β activity alteration , P05067 triplication and the defective neuronal production that characterizes the DS brain . Knowledge of the molecular mechanisms underlying neurogenesis alterations in DS may help to devise therapeutic strategies , potentially usable in humans . Results suggest that drugs that increase GSK3β phosphorylation , such as lithium or fluoxetine , may represent useful tools for the improvement of neurogenesis in DS .